靶向Survivin的RNAi联合放射治疗肺腺癌的研究
|
摘要
肺癌的发生、发展以及侵袭、转移是多基因参与、多步骤发生的过程。全世界发病率最高的癌症是肺癌,而且占全世界癌症死亡的第一位。肺癌的治疗主要包括手术、放疗和化疗,但是都具有局限性,近年来,新兴的肿瘤分子靶向治疗意在针对肿瘤内特异的分子治疗靶点,利用一定的分子技术阻断分子的生物学功能,进而从分子水平逆转肿瘤细胞的恶性生物学行为,达到抑制肿瘤生长的目的,是近年来倍受关注的领域,而将肺癌的传统治疗方法与分子靶向治疗联合应用是个非常有前景的技术手段。Survivin基因是凋亡抑制蛋白家族的重要成员,具有抑制凋亡、能参与细胞周期调控、促进血管生成的功能,有望成为肺癌靶向分子治疗的新靶点。RNA干扰(RNA interference,RNAi)技术是近年来发展起来的一项特异性抑制功能基因表达的方法,具有操作简单、抑制高效和特异性等特点,将RNAi技术引入肺癌的分子靶向治疗也是新兴起的科学研究方向。电离辐射可以直接杀伤肺腺癌A549细胞,并且诱导细胞凋亡,但是临床肺癌的放疗中多次照射可能产生辐射耐受,疗效不佳;又因为Survivin基因具有抑制凋亡的作用,笔者考虑利用RNAi技术沉默Survivin基因的表达,去除抑制凋亡的作用,在放疗时可以增加细胞的辐射敏感性,对临床的肺癌治疗提供新的思路。本实验构建了靶向Survivin基因的干扰表达载体pGenesil-2-Survivin,转染肺腺癌A549细胞,并联合X射线照射,观察Survivin mRNA及蛋白的表达,同时观察细胞增殖、周期、凋亡和放射敏感性的变化,结果显示转染pGenesil-2-Survivin表达载体后明显降低其mRNA及蛋白的表达,增加细胞凋亡百分率,减弱细胞的增殖能力,提高细胞的放射敏感性;而X射线照射能够增强细胞的这些生物学效应。进而笔者构建肺腺癌裸鼠移植模型,将pGenesil-2-Survivin表达载体局部注射到瘤体内,可以观察到Survivin mRNA及蛋白的表达明显降低,同时肿瘤生长明显抑制,而X射线照射能够增强肿瘤的抑制效果。本研究的结果显示无论是体外还是体内实验,都能证明靶向Survivin的RNAi能够提高肺腺癌的放射治疗效应,这为肺癌的分子靶向治疗提供了新治疗方案和实验证据。
Lung cancer is highest incidence of cancer in world, and accounts for the first one of the proportion of cancer deaths, the occurrence, the development, and the invasion,the metastasis of lung cancer are a multi-gene participating, multi-step developing process. The general treatment of lung cancer includes operation, radiotherapy and chemotherapy, but has limitations, in recent years, the rise of molecular targeting therapy of lung cancer are expected to combine with traditional regimen and solve thorough the therapy of lung cancer treatment. The so-called molecular targeting therapy is utilizing molecular targeting technology to block specific molecular target and its biological function, thereby reverses the biological behavior of malignant tumor cells on the molecular level, to reach purposesof tumor growth inhibition. Survivin gene is the inhibitor of apoptosis protein family member, it can inhibit apoptosis, involve in cell cycle regulation and promote angiogenesis function, is expected to become a target for cancer therapy; RNAi is a new development techniques of specific functional gene expression inhibition, using RANi technique to silence Survivin gene expression, thus to inhibit the role of Survivin in tumors can induce tumor cell apoptosis. Ionizing radiation can be directly used to treat tumors, but the long application has radiation tolerance phenomenon, and the combination of silencing Survivin gene and ionizing radiation can significantly increase the efficacy of radiation therapy. In this study, using the features of the three and combining them, to investigate the inhition effects on A549 cells and their graft tumor in vitro and in vivo after Survivin gene silencing, and to explore the mechanism. This experiment can provides a new treatment programs and experimental evidence for molecular target for clinical treatment of lung cancer.
1 Effects on A549 cell apoptosis and Survivin protein expression of ionizing radiation
Ionizing radiation can induce tumor cell apoptosis, its mechanism is very complicated, involves many genes. In this study, the apoptosis percentage of lung adenocarcinoma A549 cells 0 ~ 72 h after 0, 0.5, 1, 2.5 and 5 Gy X-ray irradiation were measured by flow cytometry, at same time, Survivin protein expression regularity in cells were detected by flow cytometry. The results showed that the apoptosis percentage of lung adenocarcinoma A549 cells 0 ~ 72 h after the same dose rradiation increased, and reached a peak at 12 or 24 h, then back down, but still were higher than the control; the percentage of apoptosis increased with dose increase same period of time after different doses irradiation, and reached the maximum 24 h after 5 Gy irradiation; at the same time, Surivin protein expression also had similar regulatity with apoptosis.Although Survivin is the apoptosis inhibition protein, but as the expression increased after radiation, apoptosis was not inhibited but actually increased, this suggested that there mignt be other factors effect them.
2 Construction of shRNA vectors targeting Survivin and it’s efficiency evaluation
In order to study lung adenocarcinoma A549 cell biological behavior change after Survivin was inhibited, the research used RNAi technique to specifically block Survivin gene expression. Two pairs of silencing sequence targeted Survivi gene were designed mainly with biological means, and linkaged to pGenesil-2 vector, to construct shRNA expression plasmid targeted Survivin, pGenesil-2-Survivin was identificated by restriction enzyme and sequencing, then transfected the expression vector into A549 cells with liposome, transfection efficiency was evalued with eukaryotic expression vector pGenesil-1 carried green fluorescence protein by flow cytometry and fluorescence microscope, and after shRNA expression plasmid was trasfected into A549 cells, Survivin mRNA and protein expressions were measured with RT-PCR and Western blot, and to evaluate interferencing efficiency. The results showed that 389 bp and 4206 bp fragments could be seen after pGenesil-2-Survivin plasmid digested by KpnⅠand EcoRⅠrestriction enzyme, this indicated that pGenesil-2-Survivin was right, the sequencing results were consistent with design, expression vector was entirely correct. cells with a green fluorescent were more than cells of normal group under fluorescence microscope, the flow cytometry results showed that green fluorescent protein expression rate was (56.22±8.16)%, transfection efficiency could reach 51.64%, the transfection efficiency could meet the experimental needs. After cells were transfected with the pGenesil2-Survivin, the Survivin mRNA and protein expression in cells were significantly inhibited, indicating Survivin gene is effectively silenced.
3 Effect on A549 cells biological behavior of shRNA vector targeted Survivin
After the shRNA targeting Survivin expression vector pGenesil-2-Survivin were transfected into A549 cells, to measure cell proliferation change by plotting cell survival curve and MTT assay; and to measure cell phage change by flow cytometry; to detect cell apoptosis by flow cytometry after PI / AnexinⅤstaining and TUNEL; to measure the change of radio sensitivity; to measure the relative gene mRNA and protein expresson by RT-PCR and Western blot. The results showed that A549 cell proliferation were inhibited in blank vector group, transfected pGenesil-2-Surivin group, 5 Gy irradiation group and transfected pGenesil-2-Surivin+5 Gy irradiation group at 0, 2, 4, 6 and 8 d, in turns, the cells in blank vector group were inhibited small, then in 5 Gy irradiation group were inhibited more, at last, in transfected pGenesil-2-Survivin plasmid and in transfected pGenesil-2-Survivin plasmid +5 Gy group were inhibited most, particularly cells inhibition in the two combined group was strongest; MTT results showed that cell proliferation in blank plasmid was basically same with normal cell proliferation from 0 h to 72 h, but cell proliferation was inhibited in transfected pGenesil-2-Survivin group and 5 Gy irradiation group, cells proliferation was inhibted most in the two combined group. Flow cytometry results showed that cells cycle phage in transfected blank vector and 5 Gy irradiation group change little, while cells percentage of G0/G1 in transfected pGenesil- 2-Survivin group significantly increased, S phase shortened, in particular, cells percentage of G0/G1in transfected pGenesil-2-Survivin+5 Gy irradiation group increased more. Flow cytometry results with PI / AnnexinⅤstaining showed that cell apoptosis percentage did not change in transfected blank vector, and increased in 5 Gy irradiation group, and increased obviously in transfected pGenesil-2-Survivin shRNA, and was more in transfected pGenesil-2-Survivin+5 Gy group. Colony forming assay results showed that the cells in transfected blank vector survival fraction changed little, while cell survival fraction in transfected pGenesil-2-Survivin decreased obviously, the D0 values of normal cells, transfected blank vector and transfected pGenesil-2-Survivin were 3.26 Gy, 3.31 Gy and 2.11 Gy, respectively, this indicated cell radiosensitivity in transfected blank vector blank changed little, but cell radiosensitivity in transfected pGenesil-2-Survivin group increased. RT-PCR, immunocytochemistry and Western blot results showed that Survivin mRNA and protein expression in the cells transfected blank vector were affected little, and Survivin mRNA and protein expression in the cells of 5 Gy irradiation group increased, but Survivin mRNA and protein expression in the cells transfected pGenesil-2-Survivin and transfected pGenesil-2-Survivin+5 Gy irradiation were inhibited significantly. Western blot results showed that caspase-3 protein in transfected blank vector group, transfected pGenesil-2-Survivin plasmid group, 5 Gy irradiation group and combination of two increased, and in combination group the caspase-3 expression was maximum. These results suggested that successfully constructed RNAi expression vector targeting Survivin gene, after pGenesil-2-Survivin transfection into A549 cells, Survivin mRNA and protein in cells were inhibited obviously, and cell proliferation was suppressed, G0/G1 cycle arrest was induced, apoptosis of cell increased and cell radiosensitivity was enhanced.
4 The inhibitory effect on A 549 cell xenografts in nude mice of targeting Survivin shRNA
To establish the lung adenocarcinoma A 549 cells xenograft tumor in nude mice model, pGenesil-2-Survivin expression vector was injected into the tumor with liposome, then the tumor was irradiated by 5 Gy, to observe tumor growth inhibition, and to explore mechanism, results showed that the graft tu mor model successfully established, after A549 cells were injected into nude mice 4 d, the mass 5 mm in diameter could be seen. The tumor growth injected by saline, pGenesil-2-Survivin shRNA, irradiated by 5 Gy, and injected by pGenesil-2-Survivin shRNA +5 Gy irradiation was significantly different, tumor growth injected saline was very rapidly, The tumor growth injected pGenesil-2-Survivin shRNA, irradiated by 5 Gy, and injected by pGenesil-2-Survivin shRNA+ 5 Gy irradiation slowed down, tumor growth irradiated by 5 Gy was inhibited relatively weakly, while tumor growth injected expression plasmid inhibition was very strong, and combination of pGenesil-2-Survivin and 5 Gy was most strong. To detect mRNA and protein expression by RT-PCR, Western blot and immunohistochemistry, results showed that 5 Gy X-rays irradiation could induce Survivin mRNA and protein increase, while the injection of pGenesil-2-Survivin plasmid and injection added by 5 Gy irradiation could inhibit Survivin mRNA and protein expression, these results suggested that after injection of pGenesil-2-Surivin, Survivin expression was inhibited, then induced tumor cell apoptosis, this might be main reason for tumor growth inhibition. At present, there is no reports about application of RNAi technology combined with radiotherapy for lung cancer at home and abroad, the experimental results, the conclusion might provid the necessary experimental data for gene therapy for adenocarcinoma of the lung.
Lung cancer is highest incidence of cancer in world, and accounts for the first one of the proportion of cancer deaths, the occurrence, the development, and the invasion,the metastasis of lung cancer are a multi-gene participating, multi-step developing process. The general treatment of lung cancer includes operation, radiotherapy and chemotherapy, but has limitations, in recent years, the rise of molecular targeting therapy of lung cancer are expected to combine with traditional regimen and solve thorough the therapy of lung cancer treatment. The so-called molecular targeting therapy is utilizing molecular targeting technology to block specific molecular target and its biological function, thereby reverses the biological behavior of malignant tumor cells on the molecular level, to reach purposesof tumor growth inhibition. Survivin gene is the inhibitor of apoptosis protein family member, it can inhibit apoptosis, involve in cell cycle regulation and promote angiogenesis function, is expected to become a target for cancer therapy; RNAi is a new development techniques of specific functional gene expression inhibition, using RANi technique to silence Survivin gene expression, thus to inhibit the role of Survivin in tumors can induce tumor cell apoptosis. Ionizing radiation can be directly used to treat tumors, but the long application has radiation tolerance phenomenon, and the combination of silencing Survivin gene and ionizing radiation can significantly increase the efficacy of radiation therapy. In this study, using the features of the three and combining them, to investigate the inhition effects on A549 cells and their graft tumor in vitro and in vivo after Survivin gene silencing, and to explore the mechanism. This experiment can provides a new treatment programs and experimental evidence for molecular target for clinical treatment of lung cancer.
1 Effects on A549 cell apoptosis and Survivin protein expression of ionizing radiation
Ionizing radiation can induce tumor cell apoptosis, its mechanism is very complicated, involves many genes. In this study, the apoptosis percentage of lung adenocarcinoma A549 cells 0 ~ 72 h after 0, 0.5, 1, 2.5 and 5 Gy X-ray irradiation were measured by flow cytometry, at same time, Survivin protein expression regularity in cells were detected by flow cytometry. The results showed that the apoptosis percentage of lung adenocarcinoma A549 cells 0 ~ 72 h after the same dose rradiation increased, and reached a peak at 12 or 24 h, then back down, but still were higher than the control; the percentage of apoptosis increased with dose increase same period of time after different doses irradiation, and reached the maximum 24 h after 5 Gy irradiation; at the same time, Surivin protein expression also had similar regulatity with apoptosis.Although Survivin is the apoptosis inhibition protein, but as the expression increased after radiation, apoptosis was not inhibited but actually increased, this suggested that there mignt be other factors effect them.
2 Construction of shRNA vectors targeting Survivin and it’s efficiency evaluation
In order to study lung adenocarcinoma A549 cell biological behavior change after Survivin was inhibited, the research used RNAi technique to specifically block Survivin gene expression. Two pairs of silencing sequence targeted Survivi gene were designed mainly with biological means, and linkaged to pGenesil-2 vector, to construct shRNA expression plasmid targeted Survivin, pGenesil-2-Survivin was identificated by restriction enzyme and sequencing, then transfected the expression vector into A549 cells with liposome, transfection efficiency was evalued with eukaryotic expression vector pGenesil-1 carried green fluorescence protein by flow cytometry and fluorescence microscope, and after shRNA expression plasmid was trasfected into A549 cells, Survivin mRNA and protein expressions were measured with RT-PCR and Western blot, and to evaluate interferencing efficiency. The results showed that 389 bp and 4206 bp fragments could be seen after pGenesil-2-Survivin plasmid digested by KpnⅠand EcoRⅠrestriction enzyme, this indicated that pGenesil-2-Survivin was right, the sequencing results were consistent with design, expression vector was entirely correct. cells with a green fluorescent were more than cells of normal group under fluorescence microscope, the flow cytometry results showed that green fluorescent protein expression rate was (56.22±8.16)%, transfection efficiency could reach 51.64%, the transfection efficiency could meet the experimental needs. After cells were transfected with the pGenesil2-Survivin, the Survivin mRNA and protein expression in cells were significantly inhibited, indicating Survivin gene is effectively silenced.
3 Effect on A549 cells biological behavior of shRNA vector targeted Survivin
After the shRNA targeting Survivin expression vector pGenesil-2-Survivin were transfected into A549 cells, to measure cell proliferation change by plotting cell survival curve and MTT assay; and to measure cell phage change by flow cytometry; to detect cell apoptosis by flow cytometry after PI / AnexinⅤstaining and TUNEL; to measure the change of radio sensitivity; to measure the relative gene mRNA and protein expresson by RT-PCR and Western blot. The results showed that A549 cell proliferation were inhibited in blank vector group, transfected pGenesil-2-Surivin group, 5 Gy irradiation group and transfected pGenesil-2-Surivin+5 Gy irradiation group at 0, 2, 4, 6 and 8 d, in turns, the cells in blank vector group were inhibited small, then in 5 Gy irradiation group were inhibited more, at last, in transfected pGenesil-2-Survivin plasmid and in transfected pGenesil-2-Survivin plasmid +5 Gy group were inhibited most, particularly cells inhibition in the two combined group was strongest; MTT results showed that cell proliferation in blank plasmid was basically same with normal cell proliferation from 0 h to 72 h, but cell proliferation was inhibited in transfected pGenesil-2-Survivin group and 5 Gy irradiation group, cells proliferation was inhibted most in the two combined group. Flow cytometry results showed that cells cycle phage in transfected blank vector and 5 Gy irradiation group change little, while cells percentage of G0/G1 in transfected pGenesil- 2-Survivin group significantly increased, S phase shortened, in particular, cells percentage of G0/G1in transfected pGenesil-2-Survivin+5 Gy irradiation group increased more. Flow cytometry results with PI / AnnexinⅤstaining showed that cell apoptosis percentage did not change in transfected blank vector, and increased in 5 Gy irradiation group, and increased obviously in transfected pGenesil-2-Survivin shRNA, and was more in transfected pGenesil-2-Survivin+5 Gy group. Colony forming assay results showed that the cells in transfected blank vector survival fraction changed little, while cell survival fraction in transfected pGenesil-2-Survivin decreased obviously, the D0 values of normal cells, transfected blank vector and transfected pGenesil-2-Survivin were 3.26 Gy, 3.31 Gy and 2.11 Gy, respectively, this indicated cell radiosensitivity in transfected blank vector blank changed little, but cell radiosensitivity in transfected pGenesil-2-Survivin group increased. RT-PCR, immunocytochemistry and Western blot results showed that Survivin mRNA and protein expression in the cells transfected blank vector were affected little, and Survivin mRNA and protein expression in the cells of 5 Gy irradiation group increased, but Survivin mRNA and protein expression in the cells transfected pGenesil-2-Survivin and transfected pGenesil-2-Survivin+5 Gy irradiation were inhibited significantly. Western blot results showed that caspase-3 protein in transfected blank vector group, transfected pGenesil-2-Survivin plasmid group, 5 Gy irradiation group and combination of two increased, and in combination group the caspase-3 expression was maximum. These results suggested that successfully constructed RNAi expression vector targeting Survivin gene, after pGenesil-2-Survivin transfection into A549 cells, Survivin mRNA and protein in cells were inhibited obviously, and cell proliferation was suppressed, G0/G1 cycle arrest was induced, apoptosis of cell increased and cell radiosensitivity was enhanced.
4 The inhibitory effect on A 549 cell xenografts in nude mice of targeting Survivin shRNA
To establish the lung adenocarcinoma A 549 cells xenograft tumor in nude mice model, pGenesil-2-Survivin expression vector was injected into the tumor with liposome, then the tumor was irradiated by 5 Gy, to observe tumor growth inhibition, and to explore mechanism, results showed that the graft tu mor model successfully established, after A549 cells were injected into nude mice 4 d, the mass 5 mm in diameter could be seen. The tumor growth injected by saline, pGenesil-2-Survivin shRNA, irradiated by 5 Gy, and injected by pGenesil-2-Survivin shRNA +5 Gy irradiation was significantly different, tumor growth injected saline was very rapidly, The tumor growth injected pGenesil-2-Survivin shRNA, irradiated by 5 Gy, and injected by pGenesil-2-Survivin shRNA+ 5 Gy irradiation slowed down, tumor growth irradiated by 5 Gy was inhibited relatively weakly, while tumor growth injected expression plasmid inhibition was very strong, and combination of pGenesil-2-Survivin and 5 Gy was most strong. To detect mRNA and protein expression by RT-PCR, Western blot and immunohistochemistry, results showed that 5 Gy X-rays irradiation could induce Survivin mRNA and protein increase, while the injection of pGenesil-2-Survivin plasmid and injection added by 5 Gy irradiation could inhibit Survivin mRNA and protein expression, these results suggested that after injection of pGenesil-2-Surivin, Survivin expression was inhibited, then induced tumor cell apoptosis, this might be main reason for tumor growth inhibition. At present, there is no reports about application of RNAi technology combined with radiotherapy for lung cancer at home and abroad, the experimental results, the conclusion might provid the necessary experimental data for gene therapy for adenocarcinoma of the lung.
引文
[1] BONSTED A, ENGESAETER B?, H?GSET A, et al. Transgene expression is increased by photochemically mediated transduction of polycation-complexed adenoviruses [J]. Gene Ther, 2004, 11(2):152-160.
[2] DéAS O, ANGEVIN E, CHERBONNIER C, SENIK A, et al. In vivo-targeted gene delivery using antibody-based nonviral vector [J]. Hum Gene Ther, 2002, 13(9):1101-1114.
[3] BERGES BK, WOLFE JH, FRASER NW. Transduction of brain by herpes simplex virus vectors [J]. Mol Ther, 2007 15(1):20-29.
[4] HIRAOKA K, KIMURA T, LOGG CR, et al. Tumor-selective gene expression in a hepatic metastasis model after locoregional delivery of a replication- competent retrovirus vetor [J]. Clin Cancer Res, 2006, 12(23):7108-7116.
[5] ISAKA Y. Gene therapy targeting kidney diseases: routes and vehicles [J]. Clin Exp Nephrol, 2006, 10(4):229-235.
[6] FOUGENROUSSE F, BARTOLI M, POUPIOT J, et al. Phenotypic correction ofα-sarcoglycan deficiency by intra-arterial injection of a muscle-specific serotype 1 rAAV vector [J]. Mol Ther, 2007, 15(1):53-61.
[7] INDRACCOLO S, HABELER W, TISATO V, et al. Gene transfer in ovarian cancer cells: a comparison between retrovial and lentiviral vectors [J]. Cancer Res, 2002, 62(21):6099-6107.
[8] SAKODA T, KASAHARA N, KEDES L, et al. Lentiviral vector-mediated gene transfer to endotherial cells compared with adenoviral and retrovial vectors [J]. Prep Biochem Biotechnol, 2007, 37(1):1-11.
[9] RAPP UR, KORN C, CETECI F, et al. MYC is a metastasis gene for non-small-cell lung cancer [J]. PLoS One, 2009, 4(6):e6029.
[10] ADJEI AA. K-ras as a target for lung cancer therapy [J]. J Thorac Oncol. 2008, 3(6Suppl 2): S160-163.
[11] HERBST RS, JOHNSON DH, MININBERG E, et al. Phase I/II trial evaluating the anti-vascular endothelial growth factor monoclonal antibody bevacizumab in combination with the HER-1/epidermal growth factor receptor tyrosine kinase inhibitor erlotinib for patients with recurrent non-small-cell lung cancer [J]. J Clin Oncol, 2005, 23(11):2544-2555.
[12] BUNN PA JR, HELFRICH B, SORIANO AF, et al. Expression of Her-2/neu in human lung cancer cell lines by immunohistochemistry and fluorescence in situ hybridization and its relationship to in vitro cytotoxicity by trastuzumab and chemotherapeutic agents [J]. Clin Cancer Res, 2001, 7(10):3239-3250.
[13] SUPINO R, PEREGO P, GATTI L, et al. A role for c-myc in DNA damage-induced apoptosis in a human TP53-mutant small-cell lung cancer cell line [J]. Eur J Cancer, 2001, 37(17):2247-2256.
[14] SONG JS. Adenovirus-mediated suicide SCLC gene therapy using the increased activity of the hTERT promoter by the MMRE and SV40 enhancer [J]. Biosci Biotechnol Biochem, 2005, 69(1):56-62.
[15]甄时建,杨举伦.以Ras信号通路为靶标的抗肿瘤治疗研究进展[J].临床与实验病理学杂志, 2008, 24(5):607-612.
[16]陈为志,郭冬梅,刘传方.恶性血液病RAS基因突变激活与Ras蛋白法尼基化抑制剂的研究进展[J].国际输血及血液学杂志, 2006, 29 (5):414-418.
[17] GAYLO AE, LAUX KS, BATZEL EJ, et al. Delayed rejection of MHC class II-disparate skin allografts in mice treated with farnesyltransferase inhibitors [J]. Transpl Immunol, 2009, 20(3):163-170.
[18] TAYLOR SA, MARRINAN CH, LIU G, et al. Combining the farnesyltransferase inhibitor lonafarnib with paclitaxel results in enhanced growth inhibitory effects on human ovarian cancer models in vitro and in vivo [J]. Gynecol Oncol, 2008, 109(1):97-106.
[19] REUTER CW, MORGAN MA, BERGMANN L. Targeting the Ras signaling pathway: a rational, mechanism-based treatment for hematologic malignancies [J]?Blood, 2000, 96(5):1655-1669.
[20] Lombardo LJ, Camuso A, Clark J, et al. Design , synthesis, and structure-activity relationships of tetrahydroquinoline-based farnesyltransferase inhibitors [J]. Bioorg Med Chem Lett, 2005, 15(7):1895-1899.
[21] EISENBERG S, HENIS YI. Interactions of Ras proteins with the plasma membrane and their roles in signaling [J]. Cell Signal, 2008, 20(1):31-39.
[22] HASHII M, FUKUDA M, NOMURA H, et al. Up-regulation of ras-GAP genes is reversed by a MEK inhibitor and doxorubicin in v-Ki-ras-transformed NIH/3T3 fibroblasts [J]. Biochem Biophys Res Commun, 2007, 356(2):374-380.
[23] NITHYA RAMNATH, ALEX ADJEI. Inhibitors of Raf kinase and MEK signaling [J]. Update on cancer therapeutics, 2007, 2(1):111-118.
[24] ROBERT F, EZEKIEL MP, SPENCER SA, et al. PhaseⅠstudy of anti- epidermal growth factor receptor antibody cetuximab in combination with radiation therapy in patients with advanced head and neck cancer [J]. J Clin Oncol, 2001, 19(13): 3234-3243.
[25] KHANZADA UK, PARDO OE, MEIER C, et al. Potent inhibition of small-cell lung cancer cell growth by simvastatin reveals selective functions of Ras isoforms in growth factor signalling [J]. Oncogene, 2006, 25(6):877-887.
[26] PAPILA C, UZUN H, BALCI H, et al. Clinical significance and prognostic value of serum sHER-2/neu levels in patients with solid tumors [J]. Med Onco, 2009, 26(2):151-156.
[27] SHEPHERD FA, RODRIGUES PEREIRA J, CIULEANU T, et al. Erlotinib in previously treated non-small-cell lung cancer [J]. N Engl J Med, 2005, 353(2):123-132.
[28] GATZEMEIER U, GROTH G, BUTTS C, et al. Randomized phaseⅡtrial of gemcitabine-cisplatin with or without transtuzumab in HER-2 positive non-small- cell lung cancer [J]. Ann Oncol, 2004, 15(1):19-27.
[29] LANGER CJ, STEPHENSON P, THOR A, et al. Transtuzumab in the treatment of advanced non-small-cell lung cancer: is there a role [J]? J Clin Oncol, 2004, 22(7):1180-1187.
[30] FUJIWARA T, GRIMM EA, MUKHOPADHYAY T, et al. A retroviral wild-type p53 expression vector penetrates human lung cancer spheroids and inhibits growth by inducing apoptosis [J]. Cancer Res, 1993, 53(18):4129-4133.
[31] FUJIWARA T, CAI DW, GEORGES RN, et al. Therapeutic effect of a retroviral wild-type p53 expression vector in an orthotopic lung cancer model [J]. J Natl Cancer Inst, 1994, 86(19): 1458-1462.
[32] ROTH JA, NGUYEN D, LAWRENCE DD, et al. Retrovirus-mediated wild-type p53 gene transfer to tumors of patients with lung cancer [J]. Nat Med, 1996, 2(9):985-991.
[33] BANDI N, ZBINDEN S, GUGGER M, et al. miR-15a and miR-16 are implicated in cell cycle regulation in a Rb-dependent manner and are frequently deleted or down-regulated in non-small cell lung cancer [J]. Cancer Res, 2009, 69(13):5553-5559.
[34] KAWABE S, ROTH JA, WILSON DR, et al. Adenovirus-mediated p16INK4a gene expression radiosensitizes non-small cell lung cancer cells in a p53- dependent manner [J]. Oncogene, 2000, 19(47):5359-5366.
[35] Fukuoka M, Yano S, Giaccone G, et al. Multi-institutional randomized phase II trial of gefitinib for previously treated patients with advanced non-small-cell lung cancer (The IDEAL 1 Trial) [J]. J Clin Oncol, 2003, 21(12):2237-2246.
[36] GIACCONE G, HERBST RS, MANEGOLD C, et al. Gefitinib in combination with gemcitabine and cisplatin in advanced non-small-cell lung cancer; a phaseⅢtrial-INTACT1 [J]. J Clin Oncol, 2004, 22 (5):777-784.
[37] HERBST RS, GIACCONE G, SCHILLER JH, et al. Gefitinib in combination with paclitaxel and carbop latin in advanced non-small-cell lung cancer: a phase III trial-INTACT2 [J]. J Clin Oncol, 2004, 22(5):785-794.
[38] KIM ES, MAUER AM, WILLIAM WN JR, et al. A phase 2 study of cetuximab in combination with docetaxel in chemotherapy-refractory/resistant patients with advanced nonsmall cell lung cancer [J]. Cancer, 2009, 15(8):1713-1722.
[39] READE CA, GANTI AK. EGFR targeted therapy in non-small cell lung cancer:potential role of cetuximab [J]. Biologics, 2009, 3:215-224.
[40] METRO G, FINOCCHIARO G, TOSCHI L, et al. Epidermal growth factor receptor (EGFR) targeted therapies in non-small cell lung cancer (NSCLC) [J]. Rev Recent Clin Trials, 2006, 1(1):1-13.
[41] ISCHIA J, PATEL O, SHULKES A, et al. Gastrin-releasing peptide: different forms, different functions [J]. Biofactors, 2009, 35(1):69-75.
[42] ROTHENBERG ML, NELSON AR, HANDE KR. New drugs on the horizon: matrix metalloproteinase inhibitors [J]. Stem Cells, 1999, 17 (4):237-240.
[43] STEWARD WP. marimastat (BB2516): current status of development [J]. Cancer Chemother Pharmacol, 1999, 43 (Supp l):S56-60.
[44] GRIFFIOEN AW. AG-3340 (Agouron Pharmaceuticals Inc) [J]. IDrugs, 2000, 3(3):336-345.
[45] ROSSI A, MAIONE P, FERRARA ML, et al. Angiogenesis Inhibitors and Vascular Disrupting Agents in Non-Small Cell Lung Cancer [J]. Curr Med Chem, 2009, 16(30):3919-3930.
[46] JOHNSON DH, FEHRENBACHERL, NOVOTNY WF, et al. Randomized phaseⅡtrial comparing bevacizumab p lus carbop latin and paclitaxelwith carbop latin and paclitaxel alone in previously untreated locally advanced ormetastatic non-small-cell lung cancer [J]. J Clin Oncol, 2004, 22(11): 2184-2191.
[47] ROSSI A, MAIONE P, COLANTUONI G, et al. Recent developments of targeted therapies in the treatment of non-small cell lung cancer [J]. Curr Drug Discov Technol, 2009, 6(2):91-102.
[48]周园园,孙艳芹,郭琳琅.小细胞肺癌阿霉素多药耐药细胞模型的建立及其与Bcl-2家族蛋白的关系[J].山东医药, 2009, 49(3):31-34.
[49]雷炜,梁军,张彦,等. Bcl-2反义核苷酸对小细胞肺癌NCI-H69放射敏感性的影响[J].青岛大学医学院学报, 2008, 44(1):15-18.
[50]何冬梅,张洹. Bcl-2反义寡核苷酸对人肺癌裸鼠移植瘤的抑制作用[J].癌症, 2006, 25(1):40-44.
[51] GLINSKY VV, KIRIAKOVA G, GLINSKII OV, et al. Synthetic galectin-3 inhibitor increases metastatic cancer cell sensitivity to taxol-induced apoptosis in vitro and in vivo [J]. Neoplasia, 2009, 11(9):901-909.
[52] CAO C, MU Y, HALLAHAN DE, et al. XIAP and Survivin as therapeutic targets for radiation sensitization in preclinical models of lung cancer [J]. Oncogene, 2004, 23(42):7047-7052.
[53] SEKINE K, TAKUBO K, KIKUCHI R, et al. Small molecules destabilize cIAP1 by activating auto-ubiquitylation [J]. J Biol Chem, 2008, 283(14):8961-8968.
[54] YANG L, MASHIMA T, SATO S, et al. Predominant suppression of apoptosome by inhibitor of apoptosis protein in non-small cell lung cancer H460 cells: therapeutic effect of a novel polyarginine-conjugated Smac peptide [J]. Cancer Res, 2003, 63(4):831-837.
[55] FRESE S, SCHüLLER A, FRESE-SCHAPER M, et al. Cytotoxic effects of camptothecin and cisplatin combined with tumor necrosis factor-related apoptosis-inducing ligand (Apo2L/TRAIL) in a model of primary culture of non-small cell lung cancer [J]. Anticancer Res, 2009, 29(8):2905-2911.
[56]张明鑫,王健生.凋亡素与肿瘤基因治疗[J].现代肿瘤医学, 2008, 16(3):476-478.
[57] HIYAMA K, HIYAMA E, SHIOKA S, et al. Telomerase activity in small-cell and non-small cell lung cancer [J]. J Natl Cancer Inst, 1995, 87(12):895-902.
[58] LI W, NI Y, TU Z, et al. Study of telomerase activity in pleural lavage fluid specimens in patients with non-small-cell lung cancer and its clinical significance [J]. Eur J Cardiothorac Surg, 2009, 36(3):460-464.
[59] HE LF, WANG YG, XIAO T, et al. Suppression of cancer growth in mice by adeno-associated virus vector-mediated IFN-beta expression driven by hTERT promoter [J]. Cancer Lett, 2009, 286(2):196-205.
[60] Dikmen ZG, Gellert GC, Jackson S, Gryaznov S, Tressler R, Dogan P, Wright WE, Shay JW. In vivo inhibition of lung cancer by GRN163L: a novel human telomerase inhibitor [J].Cancer Res, 2005, 65 (17):7866-7873.
[61] GRYAZNOV SM, JACKSON S, DIKMEN G, et al. Oligonucleotide conjugate GRN163L targeting human telomerase as potential anticancer and antimetastatic agent [J]. Nucleosides Nucleotides Nucleic Acids, 2007, 26(10-12):1577-1579.
[63]刘永全,江涛. RNA干扰与肺癌的诊断与治疗[J].四川医学, 2007, 28(3):241-243.
[64] http://nobelprize.org/nobel_prizes /medicine/laureates/2006/
[65] ZAMORE PD. Ancient pathways programmed by small RNAs [J]. Science, 2002, 296(5571): 1265-1269.
[66] ZAMORE PD, TUSCHL T, SHARP PA, et al. RNAi: double-stranded RNA directs the ATP-dependent cleavage of mRNA at 21 to 23 nucleotide intervals [J]. Cell, 2000, 101(1):25-33.
[67] WASSENEGGER M, HEIMES S, RIEDEL L, et al. RNA-directed de novo methylation of genomic sequences in plants [J]. Cell, 1994, 76(3):567-576.
[68]杜梅君,刘德培,梁植权. RNA干扰与染色质沉默-生物体内精密的网络调控机制[J].生物化学与生物物理进展, 2004, 31(3 ):204-208.
[69] PARRISH S, FLEENOR J, XU S, et al. Functional anatomy of a dsRNA trigger: differential requirement for the two trigger strands in RNA interference [J]. Mol Cell, 2000, 6(5):1077-1087.
[70] HAMMOND SM, BERNSTEIN E, BEACH D, et al. An RNA-directed nuclease mediates post-transcriptional gene silencing in Drosophila cells [J]. Nature, 2000, 404(6775):293-296.
[71] SHRIVASTAVA N, SRIVASTAVA A. RNA interference: an emerging generation of biologicals [J]. Biotechnol J, 2008, 3(3):339-353.
[72] LUO KQ, CHANG DC. The gene-silencing efficiency of siRNA is strongly dependent on the local structure of mRNA at the targeted region [J]. Biochem Biophys Res Commun, 2004,318(1):303-310.
[73] HOLEN T, AMARZGUIOUI M, BABAIE E, et al. Similar behaviour of single-strand and double-strand siRNAs suggests they act through a common RNAi pathway [J]. Nucleic Acids Res, 2003, 31(9):2401-2407.
[74]袁成良,金晓岚. RNA干扰原理及应用研究进展[J].西部医学, 2009, 21(4):664-666.
[75] BERNSTEIN E, CAUDY AA, HAMMOND SM, et al. Role for a bidentateri bonuclease in the initiation step of RNA interference [J]. Nature, 2001, 409 (6818): 363-368.
[76] YANG D, BUCHHOLZ F, HUZNG Z, et al. Short RNA duplexes produced by hydrolysis with Escherichia coli RNAse III mediate effective RNA interference in mammalian cells [J]. Proc Natl Acad Sci USA, 2002, 99(15):9942-9947.
[77] PAUL CP, GOOD PD, WINER I, et al. Effective expression of small interfering RNA in human cells [J]. Nat Biotechnol, 2002, 20(5):505-508.
[78] SUI G, SOOHOO C, AFFAR EL B, et al. A DNA vector based RNAi technology to suppress gene expression in mammalian cells [J]. Proc Natl Acad Sci USA, 2002, 99(8):5515-5520. [79 ]董婉维,孙开来,郑志红. RNA干扰技术在基因治疗中的应用进展[J].现代生物医学进展, 2009, 9(6):1174-1177.
[80] MCCAFFREY AP, MEUSE L, PHAM TT, et al. RNA interference in adult mice [J]. Nature, 2002, 418(6893):38-39.
[81] DURCAN N, MURPHY C, CRYAN SA. Inhalable siRNA: potential as a therapeutic agent in the lungs [J]. Mol Pharm, 2008, 5(4):559-566.
[82] AIGNER A. Gene silencing through RNA interference (RNAi) in vivo: strategies based on the direct application of siRNAs [J]. J Biotechnol, 2006, 124(1):12-25.
[83] SCHIFFELERS RM, XU J, STORM G, et al. Effects of treatment with small interfering RNA on joint inflammation in mice with collagen-induced arthritis [J]. Arthritis Rheum, 2005, 52(4):1314-1318.
[84] KO D, HAWKINS L, YU DC. Development of transcriptionally regulated oncolytic adenoviruses [J]. Oncogene, 2005, 24(52):7763-7774.
[85] JIN Z,GAO F,FLAGG T,et al. Tobacco-specific nitrosamine 4- (methylnitrosamino)- 1-(3-pyridyl)-1-butanone promotes functional cooperation of Bcl2 and c-Myc through phosphorylation in regulating cell survival and proliferation [J]. J BiolChem, 2004, 279(38):40209-40219.
[86]肖希龙,涂义定,李锐,等.原癌基因c-myc RNA干扰对A549细胞周期的影响[J].毒理学杂志, 2005, 19(Suppl3):203.
[87] MA LL, SUN WJ, WANG ZH, et al. Effects of silencing of mutant p53 gene in human lung adenocarcinoma cell line Anip973 [J]. J Exp Clin Cancer Res. 2006, 25(4):585-592.
[88]陈日升,齐连权,于长明,等. RNA干涉抑制肺癌细胞DNA甲基转移酶1 (DNMT1)的表达[J].军事医学科学院院刊, 2005, 29 (21):127-131.
[89] HE DM, ZHANG Y, LIU GX. Bcl-2 small hairpin RNAs enhance radiation-induced apoptosis in A549 cells [J]. Cell Biol Int, 2007, 31(11): 1442-1445.
[90]胡海燕,张洹. siRNA对肺癌细胞株NCI-H460 bcl-2基因表达的影响[J].暨南大学学报(自然科学与医学版), 2005, 26(2):174-179.
[91] CRNKOVI?-MERTENS I, MULEY T, MEISTER M, et al. The anti-apoptotic livin gene is an important determinant for the apoptotic resistance of non-small cell lung cancer cells [J]. Lung Cancer, 2006, 54(2):135-142.
[92]孙建国,廖荣霞,陈正堂,等. Livin异构体特异siRNA表达载体的构建及其在SPC-A1细胞中的稳定表达[J].肿瘤, 2006, 26(2) :139-143.
[93] ZHANG M, ZHANG X, BAI CX, et al. Inhibition of epidermal growth factor receptor expression by RNA interference in A549 cells [J]. Acta Pharmacol Sin, 2004, 25(1):61-67.
[94]游佳,赵新宇,陈县城,等.靶向EGFR的RNA干扰诱导非小细胞肺癌细胞A549凋亡的研究[J].中国肿瘤防治杂志, 2008, 15(16):1205-1209.
[95]张泳,谷仲平,周勇安,等. RNAi沉默VEGF的表达及其治疗肺癌的初步研究[J].细胞与分子免疫学杂志, 2009, 25(4):341-343, 347.
[96] LEE YJ, IMSUMRAN A, PARK MY, et al. Adenovirus expressing shRNA to IGF-1R enhances the chemosensitivity of lung cancer cell lines by blocking IGF-1 pathway [J]. Lung Cancer, 2007, 55(3):279-286.
[97]孔敏坚,董爱强,马志原,等. RNA干扰沉默IGF-ⅠR表达对非小细胞肺癌细胞生物学特性及化疗敏感性的影响[J].浙江大学学报(医学版), 2008, 37(4):373-380.
[98] AMBROSINI G, ADIDA C, ALTIER DC. A novel anti-apoptosis gene, survivin, expressed in cancer and lymphoma [J]. Nat Med, 1997, 3(8):917-921.
[99] O'CONNOR DS, GROSSMAN D, PLESCIA J, et al. Regulation of apoptosis at cell division by p34cdc2 phosphorylation of survivin [J]. Proe Natl Acad Sci USA, 2000, 97(24):13013-13107.
[100] VERDECIA MA, HUANG H, DUTIL E, et al. Structure of the human anti-apoptotic protein survivin reveals a dimeric arrangement [J]. Nat Struct Biol, 2000, 7(7):602-608.
[101] CONWAY EM, POLLEFEYT S, COMELISSEN J, et al. Three differentially expressed survivin cDNA variants encode proteins with distinct antiapoptotic functions [J]. Blood, 2000, 95(4):1435-1442.
[102] ALTIERI DC, MARCHISIO PC. Survivin apoptosis: an interloper between cell death and cell proliferation in cancer [J]. Lab Invest, 1999, 79(11):1327-1333.
[103]廖玉华,殷秀飞. Survivin的表达和功能研究[J].中国生物工程杂志, 2006, 26(5):54-57.
[104]杨建辉.凋亡抑制基因Survivin的研究进展[J].实用癌症杂志, 2003, 18(2):216-217.
[105] FORTUGNO P, WALL NR, GIODINI A, et al. Surviivin exists in immunochemically distinct subcellular pools and is involved in spindle microtubule function [J]. J Cell Sci, 2002, 115 (Pt3):575-585.
[106] LI F, AMBROSINI G, CHU EY, et al. Control of apoptosis and mitotic spindle checkpoint by survivin [J]. Nature, 1998, 396(6711):580-584.
[107]杨冬,朱尤庆,齐健. Survivin基因与PTEN蛋白在结直肠腺癌组织中的表达及临床意义[J].癌症, 2004, 23(23):306-309.
[108] CARTER BZ, MILELLA M, ALTIERI DC, et al. Cytokine-regulated expression of survivin in myeloid leukemia [J]. Blld, 2001, 97(9):2784-2790.
[109] SCHLETTE EJ, MEDEIROS LJ, GOY A, et al. Survivin expression predictspoorer prognosis in anaplastic large-cell lymphoma [J]. J Clin Oncol, 2004, 22(9):1682-1688.
[110] KONNIKOVA L, KOTECKI M, KRUGER MM, et al. Knockdown of STAT3 expression by RNAi induces apoptosis in astrocytoma cells [J]. BMC Cancer, 2003, 3:23.
[111] CHAO JI, KUO PC, HSU TS. Down-regulation of survivin in nitric oxide-induced cell growth inhibition and apoptosis of the human lung carcinoma cells [J]. Biol Chem, 2004, 279(19):20267-20276.
[112] AMBROSINI G, ADIDA C, SIRUGO G, et al. Induction of apoptosis and inhibition of cell prolilferation by survivin gene targeting [J]. Biol Chem, 1998, 273(18):11177-11182.
[113] MESRI M, WALL NR, LI J, et al. Cancer gene therapy using a survivin mutant adenovirus [J]. J Clin Invest, 2001, 108 (7):981-990.
[114] SHIN S, SUNG BJ, CHO YS, et al. An anti-apoptotic protein human survivin is a direct inhibitor of caspase-3 and -7 [J]. Biochemistry, 2001, 40(4):1117-1123.
[115] RIEDL SJ, RENATUS M, SCHWARZENBACHER R, et al. Structural basis for the inhibition of caspase-3 by XIAP [J]. Cell, 2001, 104(5):791-800.
[116] Jin Y, Wei Y, Xiong L, Yang Y, Wu JR. Differential regulation of survivin by p53 contributes to cell cycle dependent apoptosis [J]. Cell Res, 2005, 15(5):361-370.
[117] KALLIO MJ, NIEMINEN M, ERIKSSON JE. Human inhibitor of apoptosis protein survivin participates in regulation of chromosome segregation and mitotic exit [J]. FASEB J, 2001, 15(14):2721-2723.
[118] LI F, ACKERMANN EJ, BENNETT CF, et al. Pleiotropic cell-division defects and apoptosis induced by interference with survivin function [J]. Nat cell Biol, 1999, 1(8):461-466.
[119]沈波,鞠桂芝,刘扬. Survivin与X射线诱导的染色体不稳定性[J].中华放射医学与防护杂志, 2006, 26(2):129-132.
[120] SUI L, DONG Y, OHNO M, et al. Survivin expression and its correlation withcell proliferation and prognosis in epithelial ovarian tumors [J]. Int J Oncol, 2002, 21(2):315-320.
[121] KOBAYASHI K, HATANO M, OTAKI M, et al. Expression of a murine homologue of the inhibitor of apoptosis protein is related to cell proliferation [J]. Proc Natl Acad Sci USA, 1999, 96(4):1457-1462.
[122] MORI A, WADA H, NISHIMURA Y, et al. Expression of the apoptosis gene survivin in human leukemia [J]. Int J Hematol, 2002, 75(2):161-165.
[123] TRAN J, RAK J, SHEEHAN C, et al. Marked induction of the IAP family antiapoptotic proteins survivin and XIAP by VEGF in vascular endothelial cells [J]. Biochem Biophys Res Commun, 1999, 264(3):781-788.
[124]杨六成,黄宗海,陈飞,等. Survivin siRNA对人脐静脉内皮细胞的Survivin基因及蛋白表达与凋亡的作用[J].广东医学, 2009, 30(3):354-357.
[125] PAPAPETROPOULOS A, FULTON D, MAHBOUBI K, et al. Angiopoietin-1 inhibits endothelial cell apoptosis via the Akt/survivin pathway [J]. J Bioll Chem, 2000, 275 (13):9102-9105.
[126] ZHEN HN, ZHANG X, HU PZ, et al. Survivin expression and its relation with proliferation, apoptosis, and angiogenesis in brain gliomas [J]. Cancer, 2005, 104(12):2775-2783.
[127] COMA S, NOE V, LAVARINO C, et al. Use of siRNAs and antisense oligonucleotides against survivin RNA to inhibit steps leading to tumor angiogenesis [J]. Oligonucleotides, 2004, 14(2):100-113.
[128] CONTE MS, ALTIERI DC. Survivin regulation of vascular injury [J]. Trends Cardiovasc Med, 2006, 16(4):114-117.
[129] KISHI H, IGAWA M, KIKUNO N, et al. Expression of the survivin gene in prostate cancer: correlation with clinicopathological characteristics, proliferative activity and apoptosis [J]. J Urol. 2004, 171(5):1855-1860.
[130] TANAKA K, IWAMOTO S , GON G, et al. Expression of survivin and its relationship to loss of apoptosis in breast carcinomas [J]. Clin Cancer Res, 2000, 6(1):127-134.
[131] NAKANISHI K, KAWAI T, KUMAKI F, et al. Survivin expression in atypical adenomatous hyperplasia of the lung [J]. Am J Clin Pathol, 2003, 120(5): 712-719.
[132] MONZóM, ROSELL R, FELIP E, et al. A novel anti-apoptosis gene: Re-expression of survivin messenger RNA as a prognosis marker in non-small- cell lung cancers [J]. J Clin Oncol, 1999, 17(7):2100-2104.
[133] KREN L, BRAZDIL J, HERMANOVA M, et al. Prognostic significance of anti-apoptosis proteins survivin and bcl-2 in non small cell lung carcinomas: a clinicopathologic study of 102 cases [J]. Appl Immunohistochem Mol Morphol, 2004, 12(1):44-49.
[134]张朝辉,邹声泉,余彦,等.胰腺癌细胞放疗抵抗与Survivin表达间的关系[J].中国现代医学杂志, 2006, 16(2):198-200.
[135] YOSHIDA H, SUMI T, HYUN Y, et al. Expression of survivin and matrix metalloproteinases in adenocarcinoma and squamous cell carcinoma of the uterine cervix [J]. Oncol Rep, 2003, 10(1):45-49.
[136] IKEGUCHI M, LIU J, KAIBARA N. Expression of survivin mRNA and protein in gastric cancer cell line (MKN-45) during cisplatin treatment [J]. Apoptosis, 2002, 7(1):23-29.
[137] SHAPIOR GI. Preclinical and clinical development of the cyclindependent kinase inhibitor flavopiridol [J]. Clin Cancer Res, 2004, 10(12 Pt 2):4270s-4275s.
[138] FULDA S, DEBATIN KM. Sensitization for tumor necrosis factor related apoptosis inducing ligand induced apoptosis by the chemopreventive agent resveratrol [J]. Cancer Res, 2004, 64(1):337-346.
[139]朱大冕,陈全,李奕璇,等. Survivin启动子调控的EGFP基因真核表达质粒的构建及其在肺癌细胞中的特异性表达[J].中国生物制品学杂志, 2009, 22(4):348-350.
[140]陈培生,陈村龙,王慧,等. Survivin基因启动子在多种肿瘤细胞株中的转录活性研究[J].数理医药学杂志, 2009, 22(4):403-405.
[141]孙萍胡,吴云林,董文杰,等. Survivin启动子介导自杀基因与突变体融合基因真核表达载体的构建[J]. 2009, 17(11):1085-1090.
[142]张蓓,刘嘉茵,韩素萍.含Survivin启动子重组的条件复制腺病毒联合顺铂对卵巢癌株HO-8910体外作用的实验研究[J].徐州医学院学报, 2008, 28(2):79-82.
[143] SAH NK, MUNSHI A, HOBBSM, et al. Effect of downregulation of survivin expression on radiosensitivity of human epidermoid carcinoma cells [J]. Int J Radiat Oncol Biol Phys, 2006, 66(3):852-859.
[144] KHANNA N, SEN S, SHARMA H, et al. S29 ribosomal protein induces apoptosis in H520 cells and sensitizes them to chemotherapy [J]. Biochem Biophys Res Commun, 2003, 304(1):26-35.
[145] YONESAKA K, TAMURA K, KURATA T, et al. Small interfering RNA targeting survivin sensitizes lung cancer cell wit h mutant p53 to adriamycin [J]. Int J Cancer, 2006, 118(4):812-820.
[146]刘黎明,陈昌杰,陈正徐,等. Survivin特异小干扰RNA表达载体构建及对肺腺癌细胞增殖和凋亡的影响[J].基础医学与临床, 2007, 27(12): 1365-1369.
[147]曾亮,吴胜其,王晖,等. Survivin基因RNA干扰对肺癌细胞凋亡及增殖的影响[J].现代生物医学进展, 2009, 9(2):271-273.
[148] HIRSCHOWITZ EA, FOODY T, KRYSCIO R, et al. Autologous dendritic cell vaccines for Non-small-cell lung cancer [J]. J Clin Oncol, 2004, 22(14):2808- 2815.
[149] ICHIKI Y, HANAGIRI T, TAKENOYAMA M, et al. Tumor specific expression of survivin-2B in lung cancer as a novel target of immunotherapy [J]. Lung Cancer, 2005, 48 (2):281-289.
[150] MAZUMDER S, GONG B, CHEN Q, et al. Proteolytic cleavage of cycline leads to inactivation of associated kinase activity and amplification of apoptosis in hematopoietic cells [J]. Mol Cell Biol, 2002, 22(7):2398-2409.
[151] OH SH, LIM SC. A rapid and transient ROS generation by cadmium triggers apoptosis via caspase-dependent pathway in HepG2 cells and this is inhibitedthrough N-acetylcysteine-mediated catalase upregulation [J].Toxicol Appl Pharmacol, 2006,12(3):212-223.
[152] WANG H, SHANG L, HAO W. DNA-damaging reagents induce apoptosis through reactive oxygen species-dependent Fas aggregation [J]. Oncogene, 2003, 22 (50):8168.
[153] YASUMOTO J, IMAI Y, TAKAHASHI A, et al. Analysis of apoptosis-related gene expression after X-ray irradiation in human tongue squamous cell carcinoma cells harboring wild-type or mutated p53 gene [J]. J Radiat Res, 2003, 44(1):41-45.
[154] LAGARDE F, AXELSSON G, DZMBER L, et al . Residential radon and lung cancer among never-smokers in Sweden [J]. Epi demiology, 2001, 12(4):396.
[155]罗利民.电离辐射对肺癌A549细胞凋亡及细胞周期的影响[J].肿瘤学杂志, 2009, 15(5):410-411.
[156] ELBASHIR SM, HARBORTH J, LENDECKEL W, et al. Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells [J]. Nature, 2001, 411(6836):494-498.
[157]刘树铮主编.医学放射生物学.北京:原子能出版社, 2006.
[158] Howard-Till RA, Yao MC. Induction of gene silencing by hairpin RNA expression in Tetrahymena thermophila reveals a second small RNA pathway [J]. Mol Cell Biol, 2006, 26(23): 8731-8742.
[159] Moamman T. Rapid colorimetric assay for celluar growth and survial: appication to proliferation cytotoxicity assays [J]. J Immunol Methods, 1983, 65(1-2):55-63.
[160] ASANUMA K, MORIAI R, YAJIMA T, et al. Survivin as a radioresistance factor in pancreatic cancer [J]. Jpn J Cancer Res, 2000, 91(11):1204-1209.
[161] SHIN S, SUNG B J, CHO YS, et al. An anti-apoptotic protein human survivin is a direct inhibitor of caspase-3 and -7 [J]. Biochemistry, 2001, 40(4): 1117- 1123.
[162] SONG H, XIN XY, XIAO F, et al. Survivin gene RNA interference inhibitsproliferation, induces apoptosis, and enhances radiosensitivity in HeLa cells [J]. Eur J Obstet Gynecol Reprod Biol, 2008, 136(1):83-89.
[163] MIAO GY, LU QM, ZHANG XL. Downregulation of survivin by RNAi inhibits growth of human gastric carcinoma cells [J]. World J Gastroenterol, 2007, 13(8):1170-1174.
[164]司徒镇强,吴军正.细胞培养[M].西安:世界图书出版社, 2004.
[165] CARMELL MA, ZHANG L, CONKLIN DS, et al.Germlinc transmission of RNAi in mice [J]. Nat Struct Biol, 2003, 10(2):91-92.
[166] SONG E, LEE SK, WANG J, et al. RNA interference targeting Fas protects mice from fulminant hepatitis [J]. Nat Med, 2003, 9(3):347-351.
[167] LEWIS D L, HAGSTROM J E, LOOMIS A G, et al. Efficient delivery of siRNA for inhibition of gene expression in postnatal mice [J]. Nat Genet, 2002, 32(1):107-108.
[168] PADDISON PJ,CANDY AA,BERSTEIN E,et al. Short hairpin RNAs(shRNAs) induce sequence-specific silencing in mammalian cells [J]. Genes Dev, 2002, 16(8):948-958.
[2] DéAS O, ANGEVIN E, CHERBONNIER C, SENIK A, et al. In vivo-targeted gene delivery using antibody-based nonviral vector [J]. Hum Gene Ther, 2002, 13(9):1101-1114.
[3] BERGES BK, WOLFE JH, FRASER NW. Transduction of brain by herpes simplex virus vectors [J]. Mol Ther, 2007 15(1):20-29.
[4] HIRAOKA K, KIMURA T, LOGG CR, et al. Tumor-selective gene expression in a hepatic metastasis model after locoregional delivery of a replication- competent retrovirus vetor [J]. Clin Cancer Res, 2006, 12(23):7108-7116.
[5] ISAKA Y. Gene therapy targeting kidney diseases: routes and vehicles [J]. Clin Exp Nephrol, 2006, 10(4):229-235.
[6] FOUGENROUSSE F, BARTOLI M, POUPIOT J, et al. Phenotypic correction ofα-sarcoglycan deficiency by intra-arterial injection of a muscle-specific serotype 1 rAAV vector [J]. Mol Ther, 2007, 15(1):53-61.
[7] INDRACCOLO S, HABELER W, TISATO V, et al. Gene transfer in ovarian cancer cells: a comparison between retrovial and lentiviral vectors [J]. Cancer Res, 2002, 62(21):6099-6107.
[8] SAKODA T, KASAHARA N, KEDES L, et al. Lentiviral vector-mediated gene transfer to endotherial cells compared with adenoviral and retrovial vectors [J]. Prep Biochem Biotechnol, 2007, 37(1):1-11.
[9] RAPP UR, KORN C, CETECI F, et al. MYC is a metastasis gene for non-small-cell lung cancer [J]. PLoS One, 2009, 4(6):e6029.
[10] ADJEI AA. K-ras as a target for lung cancer therapy [J]. J Thorac Oncol. 2008, 3(6Suppl 2): S160-163.
[11] HERBST RS, JOHNSON DH, MININBERG E, et al. Phase I/II trial evaluating the anti-vascular endothelial growth factor monoclonal antibody bevacizumab in combination with the HER-1/epidermal growth factor receptor tyrosine kinase inhibitor erlotinib for patients with recurrent non-small-cell lung cancer [J]. J Clin Oncol, 2005, 23(11):2544-2555.
[12] BUNN PA JR, HELFRICH B, SORIANO AF, et al. Expression of Her-2/neu in human lung cancer cell lines by immunohistochemistry and fluorescence in situ hybridization and its relationship to in vitro cytotoxicity by trastuzumab and chemotherapeutic agents [J]. Clin Cancer Res, 2001, 7(10):3239-3250.
[13] SUPINO R, PEREGO P, GATTI L, et al. A role for c-myc in DNA damage-induced apoptosis in a human TP53-mutant small-cell lung cancer cell line [J]. Eur J Cancer, 2001, 37(17):2247-2256.
[14] SONG JS. Adenovirus-mediated suicide SCLC gene therapy using the increased activity of the hTERT promoter by the MMRE and SV40 enhancer [J]. Biosci Biotechnol Biochem, 2005, 69(1):56-62.
[15]甄时建,杨举伦.以Ras信号通路为靶标的抗肿瘤治疗研究进展[J].临床与实验病理学杂志, 2008, 24(5):607-612.
[16]陈为志,郭冬梅,刘传方.恶性血液病RAS基因突变激活与Ras蛋白法尼基化抑制剂的研究进展[J].国际输血及血液学杂志, 2006, 29 (5):414-418.
[17] GAYLO AE, LAUX KS, BATZEL EJ, et al. Delayed rejection of MHC class II-disparate skin allografts in mice treated with farnesyltransferase inhibitors [J]. Transpl Immunol, 2009, 20(3):163-170.
[18] TAYLOR SA, MARRINAN CH, LIU G, et al. Combining the farnesyltransferase inhibitor lonafarnib with paclitaxel results in enhanced growth inhibitory effects on human ovarian cancer models in vitro and in vivo [J]. Gynecol Oncol, 2008, 109(1):97-106.
[19] REUTER CW, MORGAN MA, BERGMANN L. Targeting the Ras signaling pathway: a rational, mechanism-based treatment for hematologic malignancies [J]?Blood, 2000, 96(5):1655-1669.
[20] Lombardo LJ, Camuso A, Clark J, et al. Design , synthesis, and structure-activity relationships of tetrahydroquinoline-based farnesyltransferase inhibitors [J]. Bioorg Med Chem Lett, 2005, 15(7):1895-1899.
[21] EISENBERG S, HENIS YI. Interactions of Ras proteins with the plasma membrane and their roles in signaling [J]. Cell Signal, 2008, 20(1):31-39.
[22] HASHII M, FUKUDA M, NOMURA H, et al. Up-regulation of ras-GAP genes is reversed by a MEK inhibitor and doxorubicin in v-Ki-ras-transformed NIH/3T3 fibroblasts [J]. Biochem Biophys Res Commun, 2007, 356(2):374-380.
[23] NITHYA RAMNATH, ALEX ADJEI. Inhibitors of Raf kinase and MEK signaling [J]. Update on cancer therapeutics, 2007, 2(1):111-118.
[24] ROBERT F, EZEKIEL MP, SPENCER SA, et al. PhaseⅠstudy of anti- epidermal growth factor receptor antibody cetuximab in combination with radiation therapy in patients with advanced head and neck cancer [J]. J Clin Oncol, 2001, 19(13): 3234-3243.
[25] KHANZADA UK, PARDO OE, MEIER C, et al. Potent inhibition of small-cell lung cancer cell growth by simvastatin reveals selective functions of Ras isoforms in growth factor signalling [J]. Oncogene, 2006, 25(6):877-887.
[26] PAPILA C, UZUN H, BALCI H, et al. Clinical significance and prognostic value of serum sHER-2/neu levels in patients with solid tumors [J]. Med Onco, 2009, 26(2):151-156.
[27] SHEPHERD FA, RODRIGUES PEREIRA J, CIULEANU T, et al. Erlotinib in previously treated non-small-cell lung cancer [J]. N Engl J Med, 2005, 353(2):123-132.
[28] GATZEMEIER U, GROTH G, BUTTS C, et al. Randomized phaseⅡtrial of gemcitabine-cisplatin with or without transtuzumab in HER-2 positive non-small- cell lung cancer [J]. Ann Oncol, 2004, 15(1):19-27.
[29] LANGER CJ, STEPHENSON P, THOR A, et al. Transtuzumab in the treatment of advanced non-small-cell lung cancer: is there a role [J]? J Clin Oncol, 2004, 22(7):1180-1187.
[30] FUJIWARA T, GRIMM EA, MUKHOPADHYAY T, et al. A retroviral wild-type p53 expression vector penetrates human lung cancer spheroids and inhibits growth by inducing apoptosis [J]. Cancer Res, 1993, 53(18):4129-4133.
[31] FUJIWARA T, CAI DW, GEORGES RN, et al. Therapeutic effect of a retroviral wild-type p53 expression vector in an orthotopic lung cancer model [J]. J Natl Cancer Inst, 1994, 86(19): 1458-1462.
[32] ROTH JA, NGUYEN D, LAWRENCE DD, et al. Retrovirus-mediated wild-type p53 gene transfer to tumors of patients with lung cancer [J]. Nat Med, 1996, 2(9):985-991.
[33] BANDI N, ZBINDEN S, GUGGER M, et al. miR-15a and miR-16 are implicated in cell cycle regulation in a Rb-dependent manner and are frequently deleted or down-regulated in non-small cell lung cancer [J]. Cancer Res, 2009, 69(13):5553-5559.
[34] KAWABE S, ROTH JA, WILSON DR, et al. Adenovirus-mediated p16INK4a gene expression radiosensitizes non-small cell lung cancer cells in a p53- dependent manner [J]. Oncogene, 2000, 19(47):5359-5366.
[35] Fukuoka M, Yano S, Giaccone G, et al. Multi-institutional randomized phase II trial of gefitinib for previously treated patients with advanced non-small-cell lung cancer (The IDEAL 1 Trial) [J]. J Clin Oncol, 2003, 21(12):2237-2246.
[36] GIACCONE G, HERBST RS, MANEGOLD C, et al. Gefitinib in combination with gemcitabine and cisplatin in advanced non-small-cell lung cancer; a phaseⅢtrial-INTACT1 [J]. J Clin Oncol, 2004, 22 (5):777-784.
[37] HERBST RS, GIACCONE G, SCHILLER JH, et al. Gefitinib in combination with paclitaxel and carbop latin in advanced non-small-cell lung cancer: a phase III trial-INTACT2 [J]. J Clin Oncol, 2004, 22(5):785-794.
[38] KIM ES, MAUER AM, WILLIAM WN JR, et al. A phase 2 study of cetuximab in combination with docetaxel in chemotherapy-refractory/resistant patients with advanced nonsmall cell lung cancer [J]. Cancer, 2009, 15(8):1713-1722.
[39] READE CA, GANTI AK. EGFR targeted therapy in non-small cell lung cancer:potential role of cetuximab [J]. Biologics, 2009, 3:215-224.
[40] METRO G, FINOCCHIARO G, TOSCHI L, et al. Epidermal growth factor receptor (EGFR) targeted therapies in non-small cell lung cancer (NSCLC) [J]. Rev Recent Clin Trials, 2006, 1(1):1-13.
[41] ISCHIA J, PATEL O, SHULKES A, et al. Gastrin-releasing peptide: different forms, different functions [J]. Biofactors, 2009, 35(1):69-75.
[42] ROTHENBERG ML, NELSON AR, HANDE KR. New drugs on the horizon: matrix metalloproteinase inhibitors [J]. Stem Cells, 1999, 17 (4):237-240.
[43] STEWARD WP. marimastat (BB2516): current status of development [J]. Cancer Chemother Pharmacol, 1999, 43 (Supp l):S56-60.
[44] GRIFFIOEN AW. AG-3340 (Agouron Pharmaceuticals Inc) [J]. IDrugs, 2000, 3(3):336-345.
[45] ROSSI A, MAIONE P, FERRARA ML, et al. Angiogenesis Inhibitors and Vascular Disrupting Agents in Non-Small Cell Lung Cancer [J]. Curr Med Chem, 2009, 16(30):3919-3930.
[46] JOHNSON DH, FEHRENBACHERL, NOVOTNY WF, et al. Randomized phaseⅡtrial comparing bevacizumab p lus carbop latin and paclitaxelwith carbop latin and paclitaxel alone in previously untreated locally advanced ormetastatic non-small-cell lung cancer [J]. J Clin Oncol, 2004, 22(11): 2184-2191.
[47] ROSSI A, MAIONE P, COLANTUONI G, et al. Recent developments of targeted therapies in the treatment of non-small cell lung cancer [J]. Curr Drug Discov Technol, 2009, 6(2):91-102.
[48]周园园,孙艳芹,郭琳琅.小细胞肺癌阿霉素多药耐药细胞模型的建立及其与Bcl-2家族蛋白的关系[J].山东医药, 2009, 49(3):31-34.
[49]雷炜,梁军,张彦,等. Bcl-2反义核苷酸对小细胞肺癌NCI-H69放射敏感性的影响[J].青岛大学医学院学报, 2008, 44(1):15-18.
[50]何冬梅,张洹. Bcl-2反义寡核苷酸对人肺癌裸鼠移植瘤的抑制作用[J].癌症, 2006, 25(1):40-44.
[51] GLINSKY VV, KIRIAKOVA G, GLINSKII OV, et al. Synthetic galectin-3 inhibitor increases metastatic cancer cell sensitivity to taxol-induced apoptosis in vitro and in vivo [J]. Neoplasia, 2009, 11(9):901-909.
[52] CAO C, MU Y, HALLAHAN DE, et al. XIAP and Survivin as therapeutic targets for radiation sensitization in preclinical models of lung cancer [J]. Oncogene, 2004, 23(42):7047-7052.
[53] SEKINE K, TAKUBO K, KIKUCHI R, et al. Small molecules destabilize cIAP1 by activating auto-ubiquitylation [J]. J Biol Chem, 2008, 283(14):8961-8968.
[54] YANG L, MASHIMA T, SATO S, et al. Predominant suppression of apoptosome by inhibitor of apoptosis protein in non-small cell lung cancer H460 cells: therapeutic effect of a novel polyarginine-conjugated Smac peptide [J]. Cancer Res, 2003, 63(4):831-837.
[55] FRESE S, SCHüLLER A, FRESE-SCHAPER M, et al. Cytotoxic effects of camptothecin and cisplatin combined with tumor necrosis factor-related apoptosis-inducing ligand (Apo2L/TRAIL) in a model of primary culture of non-small cell lung cancer [J]. Anticancer Res, 2009, 29(8):2905-2911.
[56]张明鑫,王健生.凋亡素与肿瘤基因治疗[J].现代肿瘤医学, 2008, 16(3):476-478.
[57] HIYAMA K, HIYAMA E, SHIOKA S, et al. Telomerase activity in small-cell and non-small cell lung cancer [J]. J Natl Cancer Inst, 1995, 87(12):895-902.
[58] LI W, NI Y, TU Z, et al. Study of telomerase activity in pleural lavage fluid specimens in patients with non-small-cell lung cancer and its clinical significance [J]. Eur J Cardiothorac Surg, 2009, 36(3):460-464.
[59] HE LF, WANG YG, XIAO T, et al. Suppression of cancer growth in mice by adeno-associated virus vector-mediated IFN-beta expression driven by hTERT promoter [J]. Cancer Lett, 2009, 286(2):196-205.
[60] Dikmen ZG, Gellert GC, Jackson S, Gryaznov S, Tressler R, Dogan P, Wright WE, Shay JW. In vivo inhibition of lung cancer by GRN163L: a novel human telomerase inhibitor [J].Cancer Res, 2005, 65 (17):7866-7873.
[61] GRYAZNOV SM, JACKSON S, DIKMEN G, et al. Oligonucleotide conjugate GRN163L targeting human telomerase as potential anticancer and antimetastatic agent [J]. Nucleosides Nucleotides Nucleic Acids, 2007, 26(10-12):1577-1579.
[63]刘永全,江涛. RNA干扰与肺癌的诊断与治疗[J].四川医学, 2007, 28(3):241-243.
[64] http://nobelprize.org/nobel_prizes /medicine/laureates/2006/
[65] ZAMORE PD. Ancient pathways programmed by small RNAs [J]. Science, 2002, 296(5571): 1265-1269.
[66] ZAMORE PD, TUSCHL T, SHARP PA, et al. RNAi: double-stranded RNA directs the ATP-dependent cleavage of mRNA at 21 to 23 nucleotide intervals [J]. Cell, 2000, 101(1):25-33.
[67] WASSENEGGER M, HEIMES S, RIEDEL L, et al. RNA-directed de novo methylation of genomic sequences in plants [J]. Cell, 1994, 76(3):567-576.
[68]杜梅君,刘德培,梁植权. RNA干扰与染色质沉默-生物体内精密的网络调控机制[J].生物化学与生物物理进展, 2004, 31(3 ):204-208.
[69] PARRISH S, FLEENOR J, XU S, et al. Functional anatomy of a dsRNA trigger: differential requirement for the two trigger strands in RNA interference [J]. Mol Cell, 2000, 6(5):1077-1087.
[70] HAMMOND SM, BERNSTEIN E, BEACH D, et al. An RNA-directed nuclease mediates post-transcriptional gene silencing in Drosophila cells [J]. Nature, 2000, 404(6775):293-296.
[71] SHRIVASTAVA N, SRIVASTAVA A. RNA interference: an emerging generation of biologicals [J]. Biotechnol J, 2008, 3(3):339-353.
[72] LUO KQ, CHANG DC. The gene-silencing efficiency of siRNA is strongly dependent on the local structure of mRNA at the targeted region [J]. Biochem Biophys Res Commun, 2004,318(1):303-310.
[73] HOLEN T, AMARZGUIOUI M, BABAIE E, et al. Similar behaviour of single-strand and double-strand siRNAs suggests they act through a common RNAi pathway [J]. Nucleic Acids Res, 2003, 31(9):2401-2407.
[74]袁成良,金晓岚. RNA干扰原理及应用研究进展[J].西部医学, 2009, 21(4):664-666.
[75] BERNSTEIN E, CAUDY AA, HAMMOND SM, et al. Role for a bidentateri bonuclease in the initiation step of RNA interference [J]. Nature, 2001, 409 (6818): 363-368.
[76] YANG D, BUCHHOLZ F, HUZNG Z, et al. Short RNA duplexes produced by hydrolysis with Escherichia coli RNAse III mediate effective RNA interference in mammalian cells [J]. Proc Natl Acad Sci USA, 2002, 99(15):9942-9947.
[77] PAUL CP, GOOD PD, WINER I, et al. Effective expression of small interfering RNA in human cells [J]. Nat Biotechnol, 2002, 20(5):505-508.
[78] SUI G, SOOHOO C, AFFAR EL B, et al. A DNA vector based RNAi technology to suppress gene expression in mammalian cells [J]. Proc Natl Acad Sci USA, 2002, 99(8):5515-5520. [79 ]董婉维,孙开来,郑志红. RNA干扰技术在基因治疗中的应用进展[J].现代生物医学进展, 2009, 9(6):1174-1177.
[80] MCCAFFREY AP, MEUSE L, PHAM TT, et al. RNA interference in adult mice [J]. Nature, 2002, 418(6893):38-39.
[81] DURCAN N, MURPHY C, CRYAN SA. Inhalable siRNA: potential as a therapeutic agent in the lungs [J]. Mol Pharm, 2008, 5(4):559-566.
[82] AIGNER A. Gene silencing through RNA interference (RNAi) in vivo: strategies based on the direct application of siRNAs [J]. J Biotechnol, 2006, 124(1):12-25.
[83] SCHIFFELERS RM, XU J, STORM G, et al. Effects of treatment with small interfering RNA on joint inflammation in mice with collagen-induced arthritis [J]. Arthritis Rheum, 2005, 52(4):1314-1318.
[84] KO D, HAWKINS L, YU DC. Development of transcriptionally regulated oncolytic adenoviruses [J]. Oncogene, 2005, 24(52):7763-7774.
[85] JIN Z,GAO F,FLAGG T,et al. Tobacco-specific nitrosamine 4- (methylnitrosamino)- 1-(3-pyridyl)-1-butanone promotes functional cooperation of Bcl2 and c-Myc through phosphorylation in regulating cell survival and proliferation [J]. J BiolChem, 2004, 279(38):40209-40219.
[86]肖希龙,涂义定,李锐,等.原癌基因c-myc RNA干扰对A549细胞周期的影响[J].毒理学杂志, 2005, 19(Suppl3):203.
[87] MA LL, SUN WJ, WANG ZH, et al. Effects of silencing of mutant p53 gene in human lung adenocarcinoma cell line Anip973 [J]. J Exp Clin Cancer Res. 2006, 25(4):585-592.
[88]陈日升,齐连权,于长明,等. RNA干涉抑制肺癌细胞DNA甲基转移酶1 (DNMT1)的表达[J].军事医学科学院院刊, 2005, 29 (21):127-131.
[89] HE DM, ZHANG Y, LIU GX. Bcl-2 small hairpin RNAs enhance radiation-induced apoptosis in A549 cells [J]. Cell Biol Int, 2007, 31(11): 1442-1445.
[90]胡海燕,张洹. siRNA对肺癌细胞株NCI-H460 bcl-2基因表达的影响[J].暨南大学学报(自然科学与医学版), 2005, 26(2):174-179.
[91] CRNKOVI?-MERTENS I, MULEY T, MEISTER M, et al. The anti-apoptotic livin gene is an important determinant for the apoptotic resistance of non-small cell lung cancer cells [J]. Lung Cancer, 2006, 54(2):135-142.
[92]孙建国,廖荣霞,陈正堂,等. Livin异构体特异siRNA表达载体的构建及其在SPC-A1细胞中的稳定表达[J].肿瘤, 2006, 26(2) :139-143.
[93] ZHANG M, ZHANG X, BAI CX, et al. Inhibition of epidermal growth factor receptor expression by RNA interference in A549 cells [J]. Acta Pharmacol Sin, 2004, 25(1):61-67.
[94]游佳,赵新宇,陈县城,等.靶向EGFR的RNA干扰诱导非小细胞肺癌细胞A549凋亡的研究[J].中国肿瘤防治杂志, 2008, 15(16):1205-1209.
[95]张泳,谷仲平,周勇安,等. RNAi沉默VEGF的表达及其治疗肺癌的初步研究[J].细胞与分子免疫学杂志, 2009, 25(4):341-343, 347.
[96] LEE YJ, IMSUMRAN A, PARK MY, et al. Adenovirus expressing shRNA to IGF-1R enhances the chemosensitivity of lung cancer cell lines by blocking IGF-1 pathway [J]. Lung Cancer, 2007, 55(3):279-286.
[97]孔敏坚,董爱强,马志原,等. RNA干扰沉默IGF-ⅠR表达对非小细胞肺癌细胞生物学特性及化疗敏感性的影响[J].浙江大学学报(医学版), 2008, 37(4):373-380.
[98] AMBROSINI G, ADIDA C, ALTIER DC. A novel anti-apoptosis gene, survivin, expressed in cancer and lymphoma [J]. Nat Med, 1997, 3(8):917-921.
[99] O'CONNOR DS, GROSSMAN D, PLESCIA J, et al. Regulation of apoptosis at cell division by p34cdc2 phosphorylation of survivin [J]. Proe Natl Acad Sci USA, 2000, 97(24):13013-13107.
[100] VERDECIA MA, HUANG H, DUTIL E, et al. Structure of the human anti-apoptotic protein survivin reveals a dimeric arrangement [J]. Nat Struct Biol, 2000, 7(7):602-608.
[101] CONWAY EM, POLLEFEYT S, COMELISSEN J, et al. Three differentially expressed survivin cDNA variants encode proteins with distinct antiapoptotic functions [J]. Blood, 2000, 95(4):1435-1442.
[102] ALTIERI DC, MARCHISIO PC. Survivin apoptosis: an interloper between cell death and cell proliferation in cancer [J]. Lab Invest, 1999, 79(11):1327-1333.
[103]廖玉华,殷秀飞. Survivin的表达和功能研究[J].中国生物工程杂志, 2006, 26(5):54-57.
[104]杨建辉.凋亡抑制基因Survivin的研究进展[J].实用癌症杂志, 2003, 18(2):216-217.
[105] FORTUGNO P, WALL NR, GIODINI A, et al. Surviivin exists in immunochemically distinct subcellular pools and is involved in spindle microtubule function [J]. J Cell Sci, 2002, 115 (Pt3):575-585.
[106] LI F, AMBROSINI G, CHU EY, et al. Control of apoptosis and mitotic spindle checkpoint by survivin [J]. Nature, 1998, 396(6711):580-584.
[107]杨冬,朱尤庆,齐健. Survivin基因与PTEN蛋白在结直肠腺癌组织中的表达及临床意义[J].癌症, 2004, 23(23):306-309.
[108] CARTER BZ, MILELLA M, ALTIERI DC, et al. Cytokine-regulated expression of survivin in myeloid leukemia [J]. Blld, 2001, 97(9):2784-2790.
[109] SCHLETTE EJ, MEDEIROS LJ, GOY A, et al. Survivin expression predictspoorer prognosis in anaplastic large-cell lymphoma [J]. J Clin Oncol, 2004, 22(9):1682-1688.
[110] KONNIKOVA L, KOTECKI M, KRUGER MM, et al. Knockdown of STAT3 expression by RNAi induces apoptosis in astrocytoma cells [J]. BMC Cancer, 2003, 3:23.
[111] CHAO JI, KUO PC, HSU TS. Down-regulation of survivin in nitric oxide-induced cell growth inhibition and apoptosis of the human lung carcinoma cells [J]. Biol Chem, 2004, 279(19):20267-20276.
[112] AMBROSINI G, ADIDA C, SIRUGO G, et al. Induction of apoptosis and inhibition of cell prolilferation by survivin gene targeting [J]. Biol Chem, 1998, 273(18):11177-11182.
[113] MESRI M, WALL NR, LI J, et al. Cancer gene therapy using a survivin mutant adenovirus [J]. J Clin Invest, 2001, 108 (7):981-990.
[114] SHIN S, SUNG BJ, CHO YS, et al. An anti-apoptotic protein human survivin is a direct inhibitor of caspase-3 and -7 [J]. Biochemistry, 2001, 40(4):1117-1123.
[115] RIEDL SJ, RENATUS M, SCHWARZENBACHER R, et al. Structural basis for the inhibition of caspase-3 by XIAP [J]. Cell, 2001, 104(5):791-800.
[116] Jin Y, Wei Y, Xiong L, Yang Y, Wu JR. Differential regulation of survivin by p53 contributes to cell cycle dependent apoptosis [J]. Cell Res, 2005, 15(5):361-370.
[117] KALLIO MJ, NIEMINEN M, ERIKSSON JE. Human inhibitor of apoptosis protein survivin participates in regulation of chromosome segregation and mitotic exit [J]. FASEB J, 2001, 15(14):2721-2723.
[118] LI F, ACKERMANN EJ, BENNETT CF, et al. Pleiotropic cell-division defects and apoptosis induced by interference with survivin function [J]. Nat cell Biol, 1999, 1(8):461-466.
[119]沈波,鞠桂芝,刘扬. Survivin与X射线诱导的染色体不稳定性[J].中华放射医学与防护杂志, 2006, 26(2):129-132.
[120] SUI L, DONG Y, OHNO M, et al. Survivin expression and its correlation withcell proliferation and prognosis in epithelial ovarian tumors [J]. Int J Oncol, 2002, 21(2):315-320.
[121] KOBAYASHI K, HATANO M, OTAKI M, et al. Expression of a murine homologue of the inhibitor of apoptosis protein is related to cell proliferation [J]. Proc Natl Acad Sci USA, 1999, 96(4):1457-1462.
[122] MORI A, WADA H, NISHIMURA Y, et al. Expression of the apoptosis gene survivin in human leukemia [J]. Int J Hematol, 2002, 75(2):161-165.
[123] TRAN J, RAK J, SHEEHAN C, et al. Marked induction of the IAP family antiapoptotic proteins survivin and XIAP by VEGF in vascular endothelial cells [J]. Biochem Biophys Res Commun, 1999, 264(3):781-788.
[124]杨六成,黄宗海,陈飞,等. Survivin siRNA对人脐静脉内皮细胞的Survivin基因及蛋白表达与凋亡的作用[J].广东医学, 2009, 30(3):354-357.
[125] PAPAPETROPOULOS A, FULTON D, MAHBOUBI K, et al. Angiopoietin-1 inhibits endothelial cell apoptosis via the Akt/survivin pathway [J]. J Bioll Chem, 2000, 275 (13):9102-9105.
[126] ZHEN HN, ZHANG X, HU PZ, et al. Survivin expression and its relation with proliferation, apoptosis, and angiogenesis in brain gliomas [J]. Cancer, 2005, 104(12):2775-2783.
[127] COMA S, NOE V, LAVARINO C, et al. Use of siRNAs and antisense oligonucleotides against survivin RNA to inhibit steps leading to tumor angiogenesis [J]. Oligonucleotides, 2004, 14(2):100-113.
[128] CONTE MS, ALTIERI DC. Survivin regulation of vascular injury [J]. Trends Cardiovasc Med, 2006, 16(4):114-117.
[129] KISHI H, IGAWA M, KIKUNO N, et al. Expression of the survivin gene in prostate cancer: correlation with clinicopathological characteristics, proliferative activity and apoptosis [J]. J Urol. 2004, 171(5):1855-1860.
[130] TANAKA K, IWAMOTO S , GON G, et al. Expression of survivin and its relationship to loss of apoptosis in breast carcinomas [J]. Clin Cancer Res, 2000, 6(1):127-134.
[131] NAKANISHI K, KAWAI T, KUMAKI F, et al. Survivin expression in atypical adenomatous hyperplasia of the lung [J]. Am J Clin Pathol, 2003, 120(5): 712-719.
[132] MONZóM, ROSELL R, FELIP E, et al. A novel anti-apoptosis gene: Re-expression of survivin messenger RNA as a prognosis marker in non-small- cell lung cancers [J]. J Clin Oncol, 1999, 17(7):2100-2104.
[133] KREN L, BRAZDIL J, HERMANOVA M, et al. Prognostic significance of anti-apoptosis proteins survivin and bcl-2 in non small cell lung carcinomas: a clinicopathologic study of 102 cases [J]. Appl Immunohistochem Mol Morphol, 2004, 12(1):44-49.
[134]张朝辉,邹声泉,余彦,等.胰腺癌细胞放疗抵抗与Survivin表达间的关系[J].中国现代医学杂志, 2006, 16(2):198-200.
[135] YOSHIDA H, SUMI T, HYUN Y, et al. Expression of survivin and matrix metalloproteinases in adenocarcinoma and squamous cell carcinoma of the uterine cervix [J]. Oncol Rep, 2003, 10(1):45-49.
[136] IKEGUCHI M, LIU J, KAIBARA N. Expression of survivin mRNA and protein in gastric cancer cell line (MKN-45) during cisplatin treatment [J]. Apoptosis, 2002, 7(1):23-29.
[137] SHAPIOR GI. Preclinical and clinical development of the cyclindependent kinase inhibitor flavopiridol [J]. Clin Cancer Res, 2004, 10(12 Pt 2):4270s-4275s.
[138] FULDA S, DEBATIN KM. Sensitization for tumor necrosis factor related apoptosis inducing ligand induced apoptosis by the chemopreventive agent resveratrol [J]. Cancer Res, 2004, 64(1):337-346.
[139]朱大冕,陈全,李奕璇,等. Survivin启动子调控的EGFP基因真核表达质粒的构建及其在肺癌细胞中的特异性表达[J].中国生物制品学杂志, 2009, 22(4):348-350.
[140]陈培生,陈村龙,王慧,等. Survivin基因启动子在多种肿瘤细胞株中的转录活性研究[J].数理医药学杂志, 2009, 22(4):403-405.
[141]孙萍胡,吴云林,董文杰,等. Survivin启动子介导自杀基因与突变体融合基因真核表达载体的构建[J]. 2009, 17(11):1085-1090.
[142]张蓓,刘嘉茵,韩素萍.含Survivin启动子重组的条件复制腺病毒联合顺铂对卵巢癌株HO-8910体外作用的实验研究[J].徐州医学院学报, 2008, 28(2):79-82.
[143] SAH NK, MUNSHI A, HOBBSM, et al. Effect of downregulation of survivin expression on radiosensitivity of human epidermoid carcinoma cells [J]. Int J Radiat Oncol Biol Phys, 2006, 66(3):852-859.
[144] KHANNA N, SEN S, SHARMA H, et al. S29 ribosomal protein induces apoptosis in H520 cells and sensitizes them to chemotherapy [J]. Biochem Biophys Res Commun, 2003, 304(1):26-35.
[145] YONESAKA K, TAMURA K, KURATA T, et al. Small interfering RNA targeting survivin sensitizes lung cancer cell wit h mutant p53 to adriamycin [J]. Int J Cancer, 2006, 118(4):812-820.
[146]刘黎明,陈昌杰,陈正徐,等. Survivin特异小干扰RNA表达载体构建及对肺腺癌细胞增殖和凋亡的影响[J].基础医学与临床, 2007, 27(12): 1365-1369.
[147]曾亮,吴胜其,王晖,等. Survivin基因RNA干扰对肺癌细胞凋亡及增殖的影响[J].现代生物医学进展, 2009, 9(2):271-273.
[148] HIRSCHOWITZ EA, FOODY T, KRYSCIO R, et al. Autologous dendritic cell vaccines for Non-small-cell lung cancer [J]. J Clin Oncol, 2004, 22(14):2808- 2815.
[149] ICHIKI Y, HANAGIRI T, TAKENOYAMA M, et al. Tumor specific expression of survivin-2B in lung cancer as a novel target of immunotherapy [J]. Lung Cancer, 2005, 48 (2):281-289.
[150] MAZUMDER S, GONG B, CHEN Q, et al. Proteolytic cleavage of cycline leads to inactivation of associated kinase activity and amplification of apoptosis in hematopoietic cells [J]. Mol Cell Biol, 2002, 22(7):2398-2409.
[151] OH SH, LIM SC. A rapid and transient ROS generation by cadmium triggers apoptosis via caspase-dependent pathway in HepG2 cells and this is inhibitedthrough N-acetylcysteine-mediated catalase upregulation [J].Toxicol Appl Pharmacol, 2006,12(3):212-223.
[152] WANG H, SHANG L, HAO W. DNA-damaging reagents induce apoptosis through reactive oxygen species-dependent Fas aggregation [J]. Oncogene, 2003, 22 (50):8168.
[153] YASUMOTO J, IMAI Y, TAKAHASHI A, et al. Analysis of apoptosis-related gene expression after X-ray irradiation in human tongue squamous cell carcinoma cells harboring wild-type or mutated p53 gene [J]. J Radiat Res, 2003, 44(1):41-45.
[154] LAGARDE F, AXELSSON G, DZMBER L, et al . Residential radon and lung cancer among never-smokers in Sweden [J]. Epi demiology, 2001, 12(4):396.
[155]罗利民.电离辐射对肺癌A549细胞凋亡及细胞周期的影响[J].肿瘤学杂志, 2009, 15(5):410-411.
[156] ELBASHIR SM, HARBORTH J, LENDECKEL W, et al. Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells [J]. Nature, 2001, 411(6836):494-498.
[157]刘树铮主编.医学放射生物学.北京:原子能出版社, 2006.
[158] Howard-Till RA, Yao MC. Induction of gene silencing by hairpin RNA expression in Tetrahymena thermophila reveals a second small RNA pathway [J]. Mol Cell Biol, 2006, 26(23): 8731-8742.
[159] Moamman T. Rapid colorimetric assay for celluar growth and survial: appication to proliferation cytotoxicity assays [J]. J Immunol Methods, 1983, 65(1-2):55-63.
[160] ASANUMA K, MORIAI R, YAJIMA T, et al. Survivin as a radioresistance factor in pancreatic cancer [J]. Jpn J Cancer Res, 2000, 91(11):1204-1209.
[161] SHIN S, SUNG B J, CHO YS, et al. An anti-apoptotic protein human survivin is a direct inhibitor of caspase-3 and -7 [J]. Biochemistry, 2001, 40(4): 1117- 1123.
[162] SONG H, XIN XY, XIAO F, et al. Survivin gene RNA interference inhibitsproliferation, induces apoptosis, and enhances radiosensitivity in HeLa cells [J]. Eur J Obstet Gynecol Reprod Biol, 2008, 136(1):83-89.
[163] MIAO GY, LU QM, ZHANG XL. Downregulation of survivin by RNAi inhibits growth of human gastric carcinoma cells [J]. World J Gastroenterol, 2007, 13(8):1170-1174.
[164]司徒镇强,吴军正.细胞培养[M].西安:世界图书出版社, 2004.
[165] CARMELL MA, ZHANG L, CONKLIN DS, et al.Germlinc transmission of RNAi in mice [J]. Nat Struct Biol, 2003, 10(2):91-92.
[166] SONG E, LEE SK, WANG J, et al. RNA interference targeting Fas protects mice from fulminant hepatitis [J]. Nat Med, 2003, 9(3):347-351.
[167] LEWIS D L, HAGSTROM J E, LOOMIS A G, et al. Efficient delivery of siRNA for inhibition of gene expression in postnatal mice [J]. Nat Genet, 2002, 32(1):107-108.
[168] PADDISON PJ,CANDY AA,BERSTEIN E,et al. Short hairpin RNAs(shRNAs) induce sequence-specific silencing in mammalian cells [J]. Genes Dev, 2002, 16(8):948-958.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |