NSCLC差异蛋白的筛选及CREB/p-CREB在NSCLC中的临床意义
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摘要
肺癌,特别是非小细胞肺癌发病率和死亡率正在迅速上升,而且是世界性趋势,已成为恶性肿瘤引起死亡的首要原因。目前仍缺乏有效的用于肺癌诊断和预后监测的特异性分子标志物。因此,寻找敏感性高、特异性好的肺癌分子标志物已成为肺癌研究的一个关键。随着后基因组时代的来临,从基因水平向蛋白质水平的深化也是医学研究发展的必然趋势。通过比较疾病状态与正常生理状态蛋白质表达的差异,发现和鉴定与疾病特异相关的蛋白质,也是肿瘤研究领域里的经典思路。蛋白质的磷酸化修饰调节贯穿了几乎所有的生命活动过程,因此磷酸化蛋白的研究对于理解机体的生命活动也是十分必要的。当正常细胞中蛋白磷酸化调控出现异常时,常常会引起恶性肿瘤,研究磷酸化蛋白质差异,能更好的揭示肿瘤发生机制。本实验根据这一思路,利用pathway array方法对39例非小细胞肺癌患者的癌组织及正常肺组织进行了差异蛋白的筛选,发现了5种磷酸化蛋白(p-ERK、p-CREB、p-p38、p-PDK1、p-cdc2)有异常表达,并采用了组织芯片技术及免疫组织化学方法检测了94例非小细胞肺癌患者的癌组织和肺良性肿瘤组织中CREB/p-CREB的表达情况,进一步探讨了CREB/p-CREB的表达与非小细胞肺癌患者的临床病理因素和预后的关系,为肺癌的发生机制及防治研究提供了新的线索。
It is increasing rapidly in the mortality and morbidity of lung cancer, especially the non-small cell lung cancer(NSCLC). It is a worldwide tendency, and, it has become the first etiological factor of malignant carcinomas. So far, it still lack of a specific biomarker for early diagnosis and monitoring prognosis. So it is crucial to find a biomarker for lung cancer with high sensitivity and specificity. With the coming of post-genome period, proteomics with its special thinking method and technology which analyze the alteration of intracellular protein shows its great power to solve biological problems, and it attracts more and more researchers’attention. It have become a kind of classic thread to deal with the research of tumors that with the comparison between morbid and normality, or among different stages of lung cancer, then finding and identifing some kinds of protein that have specific relationship with the disease. According to this thread, this research made a protein-difference screening to the NSCLC tissues and normal ones with the pathway array, that we examined the difference of the expression of CREB/p-CREB between the patients of NSCLC and benefit tumors with the technic of Tissue Microarray (TMA)and immunohistochemistry (IHC). Then we made a further approach to the relationship between the expression of CREB/p-CREB and TMA as long as IHC in clinical pathogen factors and prognosis. These has provided a new clue to do the research in the development, prevention and cure of the lung cancer.
Objective: To screen the protein that express differently between NSCLC and normal tissue, and make an advance research in the relationship between CREB/p-CREB and clinical pathogen factors and prognosis of lung cancer patients.
Methods: Choose 39 cases of malignant tissue in lung cancer and pairing adjacent normal tissue. Examine the expression in the wax samples of 94 cases of NSCLC and 19 cases of benefit tumors, by using pathway array, aiming at CREB/p-CREB. Make an advance approach to the relationship between CREB/p-CREB and clinical pathogen factors and prognosis of lung cancer patients.
Result:
1.We have screen out 5 kind of protein with different expression, they are p-PDK1,p-ERK,p-CREB,p-cdc2,p-p38.
2.The over expression of CREB in NSCLC was significantly higher than in benign tumor of lung(60% vs 5%, P=0.000); the over expression of P-CREB in NSCLC was non- significantly higher than in benign tumor of lung(46%vs37%, P>0.05)
3.The over expression of CREB in squamous carcinoma was significantly higher than in adenocarcinoma(P=0.000); the over expression of CREB in smokers was significantly higher than in non-smokers( P=0.013); but they were both not related to gender, age, tumor size, node status, and TNM stage.
4.Survival analysis showed that the survival of lower-expression of CREB was better than the over-expression in adenocarcinoma patients(P=0.034).
Conclusion
1 p-PDK1,p-ERK,p-CREB,p-Cdc2,p-P38 express differently in normal lung tissues.
2 CREB/P-CREB express in nuclear; CREB has an significant higer over-expression in NSCLC tissues, and relate to the attribute of pathogen as long as smoking.
3 The survival was worse in lung adenocarcinoma patients with over-expression of CREB.
It is increasing rapidly in the mortality and morbidity of lung cancer, especially the non-small cell lung cancer(NSCLC). It is a worldwide tendency, and, it has become the first etiological factor of malignant carcinomas. So far, it still lack of a specific biomarker for early diagnosis and monitoring prognosis. So it is crucial to find a biomarker for lung cancer with high sensitivity and specificity. With the coming of post-genome period, proteomics with its special thinking method and technology which analyze the alteration of intracellular protein shows its great power to solve biological problems, and it attracts more and more researchers’attention. It have become a kind of classic thread to deal with the research of tumors that with the comparison between morbid and normality, or among different stages of lung cancer, then finding and identifing some kinds of protein that have specific relationship with the disease. According to this thread, this research made a protein-difference screening to the NSCLC tissues and normal ones with the pathway array, that we examined the difference of the expression of CREB/p-CREB between the patients of NSCLC and benefit tumors with the technic of Tissue Microarray (TMA)and immunohistochemistry (IHC). Then we made a further approach to the relationship between the expression of CREB/p-CREB and TMA as long as IHC in clinical pathogen factors and prognosis. These has provided a new clue to do the research in the development, prevention and cure of the lung cancer.
Objective: To screen the protein that express differently between NSCLC and normal tissue, and make an advance research in the relationship between CREB/p-CREB and clinical pathogen factors and prognosis of lung cancer patients.
Methods: Choose 39 cases of malignant tissue in lung cancer and pairing adjacent normal tissue. Examine the expression in the wax samples of 94 cases of NSCLC and 19 cases of benefit tumors, by using pathway array, aiming at CREB/p-CREB. Make an advance approach to the relationship between CREB/p-CREB and clinical pathogen factors and prognosis of lung cancer patients.
Result:
1.We have screen out 5 kind of protein with different expression, they are p-PDK1,p-ERK,p-CREB,p-cdc2,p-p38.
2.The over expression of CREB in NSCLC was significantly higher than in benign tumor of lung(60% vs 5%, P=0.000); the over expression of P-CREB in NSCLC was non- significantly higher than in benign tumor of lung(46%vs37%, P>0.05)
3.The over expression of CREB in squamous carcinoma was significantly higher than in adenocarcinoma(P=0.000); the over expression of CREB in smokers was significantly higher than in non-smokers( P=0.013); but they were both not related to gender, age, tumor size, node status, and TNM stage.
4.Survival analysis showed that the survival of lower-expression of CREB was better than the over-expression in adenocarcinoma patients(P=0.034).
Conclusion
1 p-PDK1,p-ERK,p-CREB,p-Cdc2,p-P38 express differently in normal lung tissues.
2 CREB/P-CREB express in nuclear; CREB has an significant higer over-expression in NSCLC tissues, and relate to the attribute of pathogen as long as smoking.
3 The survival was worse in lung adenocarcinoma patients with over-expression of CREB.
引文
[1]Alberg AJ, Samet JM,et al. Epidemiology of lung cancer[J].chest,2003,123:21-49.
[2]Parkin DM,Bray F,Ferlay J,et al.Global Cancer Statistics 2002[J].CA Cancer J Clin,2005,55(2):74-108.
[3]American Canccer sociely: Cancer Facts & Figures [M]. GA: American Cancer sociely ,2007.
[4]钱桂生.肺癌流行病学与启示[J].中国临床医生,2009,37(9): 3-4.
[5]Molina JR ,Yang P,Cassivi SD,et al.Non-small cell lung cancer:epidemiology, risk factors,treatment,and survivorship[J]. Mayo Clin Proc 2008,83:584-94.
[6]Yang L, Perkin DM,et al. Estimation and projection of the national profile of cancermorlayity in chaina 1991-2005[J].Br J Cancer,2005,90:2157-2166.
[7]Waller DA. Surgery for non-small cell lung cancer new trends[J].Lung Cancer,2001, 34(2):133-136.
[8]Gharagozloo F, Margolis M,et al. Robot-assisted thoraco-scopic lobectomy for early-stage lung cancer[J]. Ann Thorac surg, 2008,85:1880-1886.
[9]Yidizeli B,Dartevelle PG,et al. Results of primary surgery with T4 Non-small cell lung cancer during a 25-year period in a single center:the benefit is worth the risk[J]. Ann Thorac surg,2008,86(4):1065-1075.
[10]Bunn PA, Thatcher N,et al. systemic trestment for advanced(stageⅢb/Ⅴ)non-small lung cancer more treatment potions more things to consider conclusion[J]. Oncologist, 2008,13(1):37-46.
[11]Olaussen KA, Dunant A,Fouret P, et al. DNA repair by ERCC1 in non-small- cell-cancer and cisplatin-based adjuvant chemotherapy[J]. N Engl J Med, 2006,355:983-991.
[12]张子瑾,程刚.分子靶向药物治疗非小细胞肺癌的临床研究进展[J].中国新药杂志,2007,5(2):158-162.
[13]Venter JC, Adams MD, Myers EW, et al. the sequence of human gonome [J].Science,2001,291:1304-1351.
[14]Wilkins M R, Sanchez J C, Gooley A A, et al. Progress with proteome projects: why all proteins expressed by a genome should be identified and how to do it[J]. Biotech Genet Eng Rev, 1996,13:19-50.
[15]Anderson N L, Anderson N G. Proteome and proteomics: new technologies, new concepts, and new words [J]. Electrophoresis, 1998,19(11):1853-1861.
[16]Service RF.Proteomics.Can Celera Do It Again[J].Science,2000, 287(5461):2136- 2138.
[17]Fenselau C, Yao XD,Reynolds K,et al. Comparative proteomics strategies that exploit mass spectrometry[J]. Abstr Pap Am Chem S, 2001,222:84-90.
[18]Li C, Chen ZC,Xiao ZQ,et al.Comparative proteomics analysis of human lung squam- ous carcinoma[J]. Biochem Bioph Res Co, 2003, 309(1):253-260.
[19]Gygi SP, Corthals GL, Zhang Y,et al. Evaluation of two-dimensional gel electrop- horesis-based proteome analysis technology[J]. Proc NatI Acad SCI U S A,2000, 97(17):9390-9395.
[20]Unlu M, Morgan M E, Minden J S. Difference gel electrophoresis: A single gel method for detecting changes in protein extracts[J]. Electrophoresis, 1997,18(11):2071-2077.
[21]李选文,陈平,梁宋平.癌症差异蛋白质组学研究中样品分离和鉴定分析技术[J].生命科学,2007,19(1):97-102.
[22]Ong S E, Mann M,et al. Mass spectrometry-based proteomic sturns quantitative[J]. NatChemBiol,2005,1(5):252-262
[23]Stulik J,Hernychova L,Prokertova S,et al.Proteome study of colorectal carcinogenesis [J].Eletrophoresis,2001,22:3019-3025.
[24]Celis JE, Gromova I,Moreira JMA,et al.Impact of proteomics on bladder cancer research[J].Phamacogenomics,2004,5(4):381-394.
[25]Celis JE, Korc M. Proteomics of disease[J]. Mol Cell Proteomics, 2005,4(4):345-351.
[26]Takikawa M, Akiyama Y,Maruyama K,et al.Proteomic analysis of a highly metastatic gastric cancer cell line using two-dimensional differential gel electrophoresis[J].Oncol Rep,2006, 16(4):705-711.
[27]Li C,Chen Z,Xiao Z, et al.Comparative proteomeics analysis of human lung squam- ous carcinoma[J].Biochem Biophys Res Commun,2003,309(1):253-260.
[28]Li C,Xiao Z,Chen Z, et al. Proteome analysis of human lung squalors carcinoma [J].proteomics,2006,6(2):547-558.
[29]Yanagiswa K,Shyr Y,Xu B,et al.proteomic pattems of tumour subsets in non-small- cell lung cancer Lancet,2003,362:433-439.
[30]Alfonso P, Catala M, Rico Morales ML, et al. Proteomic analysis of lung biopsies: Differntial protein expression profile between peritumoral and tumoral tissue[J]. Proteomics. 2004,4(2):442-447.
[31]武建国。实用临床免疫学检验[M].江苏科技技术出版社。1990:67-70.
[32]Southern EM. Detection of specific sequences among DNA frag-ments separated by gel electrophoresis. [J] Mol Biol, 1975,98(3):503-17.
[33]Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications[J]. Proc Natl Acad Sci USA, 1979;76(9): 4350-4.
[34]Kurien BT, Scofield RH et al. Multiple immunoblots after non electrophoretic bidirectional transfer of a single SDS-PAGE gel with multiple antigens[J],J Immunol Methods, 1997;205-91.
[35]Hauri HP, Bucher K, Immunoblotting with monoclonal antibody-ties Importance of the blocking solution[J]. Anal Biochem.1986(159):386-389.
[36]Lasne F.Double-blotting: a solution to the problem of nonspecific binding of sencondary antibodies in immunoblotting procedures[J]. J Immunol Methods,2003:276:233.
[37]Lowe G. Slice blotting: a method for detecting the release of immunoreactivesubstances from living brain tissue[J]. j Neurosci Methods, 1990,90:117.
[38]Sumi MG,Mathai A, Sarada C, et al. Rapid diagnosis of tuberculous meningitis by a dot immunobinding assay to detect my cobacterial antigen in cerebrospinal fluid specimens[J]. J Clin Microbiol, 1999,37: 3925.
[39]Szakai D, Schneider G, Pai T. A colony blot immunoassay to identifiy enteroinvasive Escherichia coli and Shigella in stool sample[J]. Diagn Microbiol Infect Dis, 2003,45: 165.
[40]Reese G, Schmechel D, Ayuso R, et al . Grid-immunoblotting: a fast and simple technique to test multiple allergens with small amounts of antibody for cros reactivity[J]. J Chromatogr B Biomed Sci, 2001, 756:151.
[41]Noya O, Alareon de Noya B. The multiple antigen blot assay(MABA): a simple immunoenzymatic technique for simultaneous screening of multiple antigens[J]. IMMUNOL Lett, 1998,63:53.
[42]苏雷,支修益,张毅等.非小细胞肺癌的靶向治疗[J].中华临床医学杂志,2008,2(1):16-18.
[43]Hile M, Freed AL,et al. Possibilities to improve autom ation, speed and precision of proteome anaylysis a comparison of two drmensional electrophoresis and alter natives[J]. Electrophoresis,2001,22:4035-4052.
[44]Hanash S. Disease Proteomics[J]. Nature,2003,422(6928):266-232.
[45]Cohen P. The oriins of protein phosphorylation[J]. Nat cell Biol ,2002,4(5):127-130.
[46]隋少卉,王京兰,钱小红等.磷酸化蛋白质组学分析和定量技术的研究进展.生物化学与生物物理进展,2007,34(3):240-245.
[47]David Y Zhang,Fei Ye,Ling Gao,et al. Proteomics, Pathway array and signaling network based medicine in cancer[J]. Cell Division,2009;1-16.
[48]刘革修.3-磷酸肌醇依赖性蛋白激酶1结构与功能[J].生命科学,2005,17(5):387-391.
[49]Alessi DR, James SR,Downes CP,et al. Characterization of a -3phosphoimositidedependent protein kinase which phosphorylates and activates protein kinase Ba[J].Curr Biol,1997,7:261-269.
[50]Mora A, Komander D,Alessi DR,et al. PDK1, the master regulator of AGC kinase signal transducition[J]. Semin Cell Develop Biol, 2004,15:161-170.
[51]Mauer M, Su T, Saal LH,et al. 3-phosphoinositide-dependent protein kinase-1 potentiates upstream lesions on the phosphatidylinositol 3 kinase patyway in breast carcinoma[J]. Cancer Res,2009,69(15):6299-6306.
[52]Lu Z,Cox-Hipkin MA,WINDSOR WT, Boyapati A,et al. 3-phosphoino- sitide-dependent protein kinase-1 regulates proliferation and survival of cancer cells with an activated mitogen activated protein kinase pathway[J]. Cancer Res, 2010,8(3):421-432.
[53]Dasmahapatra GP, Didolkar P, Alley MC, et al. In vitro combination treatment with perifosine and UCN-01 demonstrates synergism against prostate(PC-3) and lung(A549) epithelial adenocarcinoma cell lines[J]. Clin Cancer Res,2004,10:5442-5252.
[54]Nurse P, Thuriaux P, Nasmyth K,et al. Genetic control of the cell division cycle in the fission yeast schizosaccharomyces pombe[J]. Mol Gen Genet,1976;146(2):167-178.
[55]翟德忠,黄强.细胞周期蛋白依赖性激酶cdc2与恶性肿瘤发生发展的研究.肿瘤,2006,26(5):489-491.
[56]Hunter T, Pines J,et al. Cyclins and cancer:cyclin D and CDK inhibitors come of age[J].cell ,1994,79;573-582.
[57]Kawamoto H, Koizumi H, Uchikoshi T,et al. Expression of the G2-M checkpoint regulators cyclin B1 and cdc2 in non-malignant and malignant human breast lesions immunocyto chemical and quantitative image analyses[J]. Am J pathol,1997,150(1):15-23.
[58]Hansel DE,Dhara S, Huang RC, et al .cdc2/CDK1 expression in esophageal adenoca- rcinoma and precursor lesions serves an a diagnostic and cancer progression marker and potential novel drug target[J]. Am J surg Pathol,2005,29(3):390-399.
[59]Ito Y, Taleda T, Wakasa K,et al. expression of the G2-M modulators in pancreaticadenocarcinoma[J]. Pancreatology, 2002,2(2):138-145.
[60]Kallakury BV, Sheehan CE,Ambros RA,et al.the prognostic significance of p34cdc2 and cyclin D1 protein expression in adenocarcinoma[J].cancer,1997,80(4):753-763.
[61]李国生,张道荣,刘冬等. P34cdc2在非小细胞肺癌中的表达及意义[J].中国医科大学学报,2002,31(2):88-92.
[62]Senju M,Sueoka N,Sato A,et al. Hsp90 inhibitors cause G2/M arrest associated with the reduction of cdc25c and cdc2 in lung cancer cell lines[J]. J cacer Res Clin oncol,2006, 132(3):150-158.
[63]Kyriakis JM,Avruch J,et al. Mammalian mitogen activated protein kinase signal transduction pathways activated by stress and inflammation[J].Physiol Rev,2001,81(2): 807-869.
[64]Brewster JL,De Valori T, Dwyer N,et al. An osmosensing signal trans duction pathway in yeast[J]. Science, 1993,259(5099):1760-1763.
[65]Han J, Lee JD, Bibbs L,et al. A MAP kinase targeted by endotoxin and hyperosmolarity in mammalian cells[J]. Science,1994,265(5173): 808-811.
[66]Obata T, Brown GE, Yaffe MB,et al. MAP kinase pathways activated by stress:the P38MAPK patyway. Crit Care Med,2000,28(4):67-77.
[67]Lee JC, Kumar S,et al. Inhibition of P38 MAP kinase as a therapeutic stategy[J]. Immunopharmacology,2000,47(2):185-201.
[68]张文军,李荣上.P38丝裂原活化蛋白激酶通路及其研究进展[J].医学综述,2007,13(9),663-665.
[69]Ostrander JH, Daniel AR, Lofgren K,et al. Breat tumor kinase(protein tyrosine kinase 6) regulates heregulin induced activation of ERK5 and P-38 MAP kinases in breast cancer cells[J]. Cancer Res, 2007,67(9):4199-4209.
[70]Atsumi T, Kato K, Uno K,et al. Pathophysiological role of the activation of p38 mitogen activated protein kinases in poorly differentiated gastric cancer[J]. Pathol Int, 2007,57(10):635-644.
[71]Junttila MR,Jokilehto T,et al. p38 alpha and p38deltami-togen-activated protein kinase isofroms regulate invasion an growth of head and neck suqamouse carcinoma cells[J]. Oncogene,2007,26(36):5267-5279.
[72]Looda K, Sasaki Y, Horimoto M,et al. Involvement of the p38 mitogen-activated protein kinase cascade in hep-atocellular carcinoma[J]. Cancer, 2003,97(12):3017-3026.
[73]王志强,谢宗涛,陈丽达等.磷酸化p38在非小细胞肺癌中表达的意义[J].肿瘤防治研究,2008,35(6):406-410.
[74]Greenberg AK, Basu S,Hu J, et al. selective p38 activation in human non-small cell lung cancer[J]. Am J Respir Cell Mol Biol,2002,26(5):558-564.
[75]Sturgill TW, Ray BL,et al. Muscle proteins related to microtubule associated protein-2 and sub-strates for and insulin-stimulatable kinase[J]. Biochem Biophys Res Commun, 1986,134(2):565-571.
[76]Boulton TG, Yancopoulos GD, Gregory JS,et al. An insulin-stimu-lated protein kinade similar to yeast kinases involved in cell cycle contro[J]. Sciences,1990,249(4964):64-67.
[77]Gary L.Johnson, Razvan Lapadat,et al. The protein kinase complement of the human genome[J]. Science,2002;296(6):1911-1912.
[78]Robinson MJ, Cobb MH,et al.Mitogen activated protein kinase pathways[J]. Curr Opin Cell Biol,1997,9(2):180-186.
[79]Ito Y, Sasaki Y, Horimoto M,et al. Activation of mitogen-activated protein kinases/extracellular signal-regula ted kinases in human hepatocellular carcinoma[J]. Hepato- logy,1998,27(4):951-958.
[80]Adeyinka A, Kui Y,Cherlet T,et al. Activated mitogen-activated protein kinase expression during human breast tumorigenesis and breast cancer progression[J]. Clin Cancer Res,2002,8(6):1747-1753.
[81]谷艳娇,包翠芬,高志安等.ERK1/2和p-ERK1/2在肺癌组织中的表达及意义[J].中国组织化学与细胞化学杂志.2009.18(3):328-331.
[82]Blackhall FH, Pintilie M, Michael M, et al. Expression and prognostic significance of kit, Protein kinse B,and mitogen-activated protein kinase in patients with small cell lung cancer[J]. Clin Cancer RES, 2003,9(6):2241-2247.
[83]Montrminy MR,sevarino KA,Wagner JA, et al.Identification of a cyclic AMP responsive element within the rat somatostatin gene[J].Proc Natl Acad Sic USA,1986, 83:6682-6686.
[84]徐瑞元,王燕峰,徐长发. CREB研究进展[J].中国生物工程学. 2003,23(1):39-47.
[85]Kathleen M. Sakamoto, David A. Frank. CREB in the pathophysiology of Cancer: Implications for Targeting Transcription Factors for Cancer Therapy[J]. Clin Cancer Res 2009,15(8):2583-2587.
[86]Kathleen M, Sakamoto, David A, et al. CREB in the Pathophysiology of cancer:lim- plications for targetin transcription factors for cancer therapy[J]. Clin cancer Res,2009,15(8): 2583-2587.
[87]Sofi M, Young MJ, Papamakarios T, et al. Role of CRE-binding protein (CREB) in aromatase expression in breast adipose[J]. Breast Cancer Res Treat 2003,79: 399–407.
[88]Unni E, Sun S, Nan B, et al. Changes in androgen receptor nongenotropic signaling correlate with transition of LNCaP cells to androgen independence[J]. Cancer Res 2004,64: 7156–68.
[89]Crans-Vargas HN, Landaw EM, Bhatia S, et al. Expression of cyclic adeno-sine monophosphate response-element binding protein in acute leukemia[J]. Blood 2002,99: 2617–2619.
[90]Hye-sook seo,Dianei D Liu, Ja seok koo.et al. Cyclic AMP Response Element- Binding Protein Over expression : A Feature Associated with Negative prognosis in Never Smokers with NSCLC[J]. Cancer Res 2008,68(15):6065-6073.
[91]Sita Aggarwal, Seung-Wook Kim, Ja Seok Koo, et al. Growth Suppression of LungCancer Cells by Targeting Cyclic AMP Response Element-Binding Protein[J]. Cancer Res 2008:68:(4),981-988.
[92]Seaki K, You A, Takaku F,et al. cell-cycle-regulated phosphorylation of cAMP response element-bunding protein:identification of nover phosphorylation sites[J].Biochem J,1999,338(1):49-54.
[93]Sutherland EW,Robison GA.,et al. The role of cyclic-3`5`-AMP in responses to catecholamines and other hormones[J]. Pharmaco Rev,1966,18(1):145-161.
[94]Montrminy MR,Bilezikjan LM. Binding of a nuclear protein to the cyclic AMP response element of the somatostatin gene[J]. Nature, 1987,328:175-178.
[95]Yamamoto KK,Gonzalez GA,Biggsl WH,et al.Phosphorylation induced binding and transcriptional efficacy of factor CREB[J].Nature,1988,334:494-498.
[96]Gonzalez GA,Yamamoto kk,Fischer WH,et al. A cluster of phosphorylation sites on the cyclic AMP regulated nuclear factor CREB predicted by its sequence[J]. Nature, 1989, 337:749-752.
[97]Berkowitz LA, Gliman MZ, et al. Two distinct forms of active transcription factor CREB(c AMP response element binding protein)[J],Proc Natil Acad Sci U S A,1990, 87(14):5258-5262.
[98]Ruppert S,Cole TJ, Boshart M, Schmid E,schutz G. Multiple mRNA isoforms of the transcription activator protein CREB:generation by altemative splicing and specific expression in primary spermatocytes[J]. EMBO J,1992,11(4):1503-1512.
[99]Gonzalez GA, Montminy MR.Cyclic cAMP stimulates somatostatin gene transcription by phosphorylaton of CREB at ser 133[J]. Cell 1989,59(4):675-680.
[100]Lee CQ, Yun Y, Hoeffler JP, et al. Cyclic AMP responsive transcription activation of CREB-327 in volves interdependent phosphorylated subdomains[J]. EMBOJ ,1990,9(13): 4445-4465.
[101]Kevin AW, Noma M ,et al. Transcription regulation by CREB and its relatives[J].Biochim Biophys Acta,1993,1174:221-233.
[102]Craig JC,Schumacher MA,Mansoor SE ,et al.Consensus and variant cAMP- regulated enhancers have distinct CREB-binding properties[J].J Biol Chem.2001, 276(15): 11719-28.
[103] Ribeiro A, Brown A, Lee KA. An in vino assay for members of the Camp response element binding protein family of transcription factors[J].1994,269(49):31124-31128.
[104]窦爱霞,董建华.cAMP信号途径和基因表达调控[J].上海第二医科大学学报,2004,24(12):1070-1073.
[105]Saeki K, You A et al. Distinct involvement of cAMP response element dependent transcriptions in functional and morphological maturation during retinoid mediated human myeloid differentiation[J]. Journal of leukocyte biology,2003,73(5):673-681.
[106]Sudzinski GP, Harrison LE, et al. Differentiation-related changes in the cell cycle traverse[J]. Int Rev Cytol, 1999,189:1-58.
[107]Lee HJ,Mignacca RC, sakamoto KM. Transcriptional activation of egr-1 by granulocyte macrophage colony stimulating factor but not interleukin 3 requires phosphory- lation of cAMP response element binding protein(CREB) on serine 133[J]. J Biol Chem,1995,270(27):15979-15983.
[108]Haywitz A, Greenbeng M. CREB: a stimulus induced transcription factor activated by a diverse array of extracellular signals[J]. Annual Review of Biochemistry, 1999,68: 821-861.
[109]Gonzalez GA, yamamoto KK, Fischer WH, et al. A cluster of phosphorylation sites on the cyclic AMP-regulated nuclear factor CREB predicted by its sequence[J]. Nature 1989;337:749-752.
[111]Swanson DJ, Zellmer E, Elaine JL. The homeodomain protein Arix interacts synergistically With cyclic AMP to regulate expression of neurotransmitter biosynthetic genes[J]. J Biol Chem,1997,272(43): 27382-27392.
[112]Matthews RP, Guthrie CR, Wailes LM et al. Calcium/calmodulin-depen-dent protein kinase typesⅡandⅣdifferentially regulate CREB-dependent gene expression[J]. Mol cell boil 1994;14:6107-6116J.
[113]Sun P, Enslen H, Myung PS, Maurer RA. Differential activation of CREB by Ca2+/ calmodulin-depen-dent protein kinase typeⅡand typeⅣinvolves phosphor-ylation of a site that negatively regulates activity[J]. Genes Dev 1994;8:2527-2539.
[114]Heist EK, Srinvasan M, Schulman H. Phosphorylation at the nuclear localization signal of Ca2+/calmodulin dependent protein kinaseⅡblocks its nuclear targeting[J]. J Bilo Chem, 1998,273(31):19763-19771.
[115]Du K, Montminy M. CREB is a regulatory target for the protein kinase AKT/PKB[J]. J Bilo Chem 1998,273:32377-32379.
[116]Philipp V,Carsten B, Bernd Nu,et al. Assigning Funcional Domains within the P101 Regulatory subunit of phosphoinositide 3 kinase[J]. J Biol Chem,2005,280:5121-5127.
[117]Murthy SS, Tosolini A,Taguchi T, et al. Mapping of AKT, encoding a member of the Akt/Protein Kinase M family, to human and rodent chromosomes by fluorescence in situ hybridization[J] . Cytogenet Cell Genet,2000,88:38-40.
[118]Downward J. Mechanisms and consequences of activation of protein kinaseB/AKT [J].Curr Opin Cell Biol,1998,10(2):262-267.
[119]Chepurny OG, Hussain MA, Holz GG,et al. Exendi-4 as a stimulator of rat insulin I gene promoter activity via bZIP/CRE interactions sensitive to serine/threonine protein kinase inhibitor RO 31-8220[J]. Endocrinology 2002; 143;2303-2313.
[120]Tan Y, Rouse J, Zhang A comb MJ,et al.Fgf and stress regulate CREB and ATF-1 via a pathway involing P38 MAP kinase and MAPKAP kinase-2[J]. EMBO J 1996;15:4629-4642.
[121]Force T, Bonventre JV,et al. Growh factors and mitogen activated protein kinases[J]. Hypertension,1998,21:152-161.
[122]Roux pp, Richards SA,Blenis J,et al. Phosphorylation of P90 ribosomal S6 kinase(RSK) regulates extracellular signal regulated kinase docking and RSK activity[J]. Mol Cell Biol,2003,23(14):4796-4804.
[123]Scheid MP, Foltz IN, Young PR, Schrader JW et al. Ceramide and cyclic adenosine monophosphate(cAMP) induce cAMP response element binding protein phosphorylation via distinct signaling pathways while having opposite effects on myeloid cell surviva[J]. Blood 1999; 93:217-225.
[124]Yamamoto KK, Gonzalez GA, Montminy MR, et al. phosphorylation induced binding and transcriptional efficacy of nuclear factor CREB[J]. Nature 1988, 334:484-498.
[125]Zanassi p, paolillo M,feliciello A et al. cAMP dependent protein kinase induces cAMP response element binding protein phosphorylation via an intracellular calcium release/ERK dependent pathway in striatal neurons[J]. J boil chem,2001,276(15):11487-11495.
[126]Mayr BM, Cancetteri G, Montminy MR,et al. Distinct effects of cAMP and mitogenic signals on CREB-binding protein recruitment impart specificity to target gene activa-tion via CREB[J]. Proc Natl Acad Sci USA,2001,98:10936-10941.
[127]Kee BL, Arias J, Montminy MR,et al. Adaptor-mediated recruitment of RNA polymeraseⅡto a signal-dependent activator[J]. J boil Chem 1996,271(5):2373-2375.
[128]Nakajima T, Uchida C, Anderson SF,Montminy MR,et al. Analysis of a cAMP responsive activator reveals a two-component mechanism for transcriptional induction via signal dependent factors[J]. Genes Dev,1997,11(6):738-747.
[129]Schiliz RL, Mizzen CA, Vassilev A, et al. Overlapping but distinct pattems of histone acetylation by the human coactivators p300 and PGAF with in nucleosomal substrates[J]. J Biol CHEM, 1999,274(3):1189-1192.
[130]Kathleen M, Sakamoto, David A, et al. CREB in the Pathophysiology of cancer:limplications for targetin transcription factors for cancer therapy[J]. Clin cancer Res,2009, 15(8): 2583-2587.
[131]Montarolo PG, Kandel ER, Schacher S. Long term heterosynaptic inhibition in(RSK) regulates extracellular signal regulated kinase docking and RSK activity[J]. Mol Cell Biol,2003,23(14):4796-4804.
[123]Scheid MP, Foltz IN, Young PR, Schrader JW et al. Ceramide and cyclic adenosine monophosphate(cAMP) induce cAMP response element binding protein phosphorylation via distinct signaling pathways while having opposite effects on myeloid cell surviva[J]. Blood 1999; 93:217-225.
[124]Yamamoto KK, Gonzalez GA, Montminy MR, et al. phosphorylation induced binding and transcriptional efficacy of nuclear factor CREB[J]. Nature 1988, 334:484-498.
[125]Zanassi p, paolillo M,feliciello A et al. cAMP dependent protein kinase induces cAMP response element binding protein phosphorylation via an intracellular calcium release/ERK dependent pathway in striatal neurons[J]. J boil chem,2001,276(15):11487-11495.
[126]Mayr BM, Cancetteri G, Montminy MR,et al. Distinct effects of cAMP and mitogenic signals on CREB-binding protein recruitment impart specificity to target gene activa-tion via CREB[J]. Proc Natl Acad Sci USA,2001,98:10936-10941.
[127]Kee BL, Arias J, Montminy MR,et al. Adaptor-mediated recruitment of RNA polymeraseⅡto a signal-dependent activator[J]. J boil Chem 1996,271(5):2373-2375.
[128]Nakajima T, Uchida C, Anderson SF,Montminy MR,et al. Analysis of a cAMP responsive activator reveals a two-component mechanism for transcriptional induction via signal dependent factors[J]. Genes Dev,1997,11(6):738-747.
[129]Schiliz RL, Mizzen CA, Vassilev A, et al. Overlapping but distinct pattems of histone acetylation by the human coactivators p300 and PGAF with in nucleosomal substrates[J]. J Biol CHEM, 1999,274(3):1189-1192.
[130]Kathleen M, Sakamoto, David A, et al. CREB in the Pathophysiology of cancer:limplications for targetin transcription factors for cancer therapy[J]. Clin cancer Res,2009, 15(8): 2583-2587.
[131]Montarolo PG, Kandel ER, Schacher S. Long term heterosynaptic inhibition in
[142]Unni E, Sun S, Nan B, et al. Changes in androgen receptor nongenotropic signaling correlate with transition of LNCaP cells to androgen independence[J]. Cancer Res 2004, 64:7156-7168.
[143]Crans vargas NH, Landaw EM, Bhatia S,et al.Expression of cyclic adeno-sime monophosphate response-element binding protein in acute leukemia[J]. Blood 2002, 99: 2617-2619.
[144]Shankar DB,Cheng JC, Kinjo K,et al. the role ofCREB as a proto-oncogene in hematopoiesis and in acute myeloid leukemia[J]. Cancer Cell.2005,7:351-362.
[145]Linnerth NM, Baldwin M, Campbell C,Moorehead RA,et al. IGF-Ⅱinduces CREB phosphorylation and cell survival in human lung cancer cells[J]. Oncogene 2005,24: 7310-1319.
[146]Hye-Sook Seo, Diane D Liu, Ja Seok Koo, et al. Cyclic AMP response element binding protein overexpression: A Feature associated with negative prognosis in never smokers with non small cell lung cancer[J].Cancer Res 2008,68(15):6065-6073.
[147]Kononen J,Bubendorf L,Kallioniemi A, et al.Tissue microarrays for high- throughput molecular profiling of tumor specimens[J].Nature 1998,4:844.
[148]王新允,朱丛中,刘婷等.组织芯片研制的几点体会[J].天津医科大学学报,2005,11(1),24-25.
[149]周小鸽,王鹏,刘金香等.组织芯片的基本应用范围.诊断病例学杂志;2003,10(6),350-352.
[150]Parkin DM,Muir CS,Whelan SL,et al.Cancer Incidence in Five Continents,vols,ⅠtoⅧ[R].Lyon:IARC, 2005.
[151]Lonescu AM, Schwarz EM, Vinson C, et al. PTHrP modulates chondrocyte differentiation through AP-1 and CREB signaling[J]. J Biol Chem 2001,276:11639-11647.
[152]Conkright MD, Montminy M. CREB:the unindicted cancer co-conspirator[J]. Trends Cell Biol 2005,15:457-459.
[153]Shankar DB,S akamoto KM.the role of cyclic-AMP binding protein(CREB) in leukemia cell prolife ration and acute leukemias[J]. Leuk Lymphoma 2004, 45:265-270.
[154]Oike Y, Takakura N,Hata A,et al.Mice homozygous for a truncated form of CREB binding protein exhibit defects in hematopoiesis and vasculo-angiogenesis[J]. Blood 1999,93:2771-2779.
[155]Stylianou E, Saklatvala J. Interleukin-1[J]. Int J Biochem cell boil,1998,30: 1075-1079.
[156]Nyormoi O, Bar-Eli M. Transcriptional regulation of metastasis-related genes in human melanoma[J]. Clin Exp Metastasis 2003,20:2 51-263.
[157]Annan RS, Huddleston MJ,verna R,et al. A multidimensional Electrospray MS-Based Approach to phosphopeptide Mapping[J]. Anal Chem,2001, 73(3):393-404.
[158]Bodenmiler B, Mueller LN, Mueller M,et al. Reproducible isolation of distinct overlapping segments of the posphoproteome[J]. Nat Methods.2007,4(3):231-237.
[159]Sita Aggarwal, Seung Wook Kim, Ja Seok Koo, et al. Growth suppression of lung cancer cells by targeting Cyclic AMP Response Element Binding Protein[J]. Cancer Res 2008, 68(4),981-988.
[160]Sharma H,Sen S, Mathur M,et al.Combined evaluation of expression of telomerase,surviving,and anti apoptotic BCL-2 family members in relation to loss of differentiation and apoptosis in human bead and neck cancers[J]. Head Neck, 2004,26 (8):733-740.
[161]Stepulak A,sifringer M,Rzeski W,et al.AMPA antagonists inhibit the extracellular signal regulated kinase athway and suppress lung cancer growth[J]. Cancer Biol Ther 2007,6:1908-1915.
[162]Shouwei Han, Neil Sidell, Susanne Roser Page,et al. Fibronectin stimulates human lung cancer carcinoma cell growth by inducing cyclooxygenase-2(cox-2) expression[J].Cancer, 2004;111:322-331.
[163]Au JS, Cho WC,Law SC,et al. Protemic approach to biomarker discovery in cancer tissue from lung adenocarcinoma among nonsmoking Chinese women in Hong Kong[J].Cancer Invest 2008, 26:128-135.
[164]Hildegrad M, Maria Cekanova,et al.NNK-induced hamster lung adenocarcinomas over expressβ-adrenergic and EGFR signaling pathways[J].lung cnacer,2005,49:35-45.
[165]Goldkon T,Khan E,et al. EGF receptor expose to cigarette smoke undergoes aberrant activation and trafficking. American Association for Cancer Ressearch Annual Meeting: Proceedings, 2009,Denver CO.phila-delphia,pa:AACR 2009.
[2]Parkin DM,Bray F,Ferlay J,et al.Global Cancer Statistics 2002[J].CA Cancer J Clin,2005,55(2):74-108.
[3]American Canccer sociely: Cancer Facts & Figures [M]. GA: American Cancer sociely ,2007.
[4]钱桂生.肺癌流行病学与启示[J].中国临床医生,2009,37(9): 3-4.
[5]Molina JR ,Yang P,Cassivi SD,et al.Non-small cell lung cancer:epidemiology, risk factors,treatment,and survivorship[J]. Mayo Clin Proc 2008,83:584-94.
[6]Yang L, Perkin DM,et al. Estimation and projection of the national profile of cancermorlayity in chaina 1991-2005[J].Br J Cancer,2005,90:2157-2166.
[7]Waller DA. Surgery for non-small cell lung cancer new trends[J].Lung Cancer,2001, 34(2):133-136.
[8]Gharagozloo F, Margolis M,et al. Robot-assisted thoraco-scopic lobectomy for early-stage lung cancer[J]. Ann Thorac surg, 2008,85:1880-1886.
[9]Yidizeli B,Dartevelle PG,et al. Results of primary surgery with T4 Non-small cell lung cancer during a 25-year period in a single center:the benefit is worth the risk[J]. Ann Thorac surg,2008,86(4):1065-1075.
[10]Bunn PA, Thatcher N,et al. systemic trestment for advanced(stageⅢb/Ⅴ)non-small lung cancer more treatment potions more things to consider conclusion[J]. Oncologist, 2008,13(1):37-46.
[11]Olaussen KA, Dunant A,Fouret P, et al. DNA repair by ERCC1 in non-small- cell-cancer and cisplatin-based adjuvant chemotherapy[J]. N Engl J Med, 2006,355:983-991.
[12]张子瑾,程刚.分子靶向药物治疗非小细胞肺癌的临床研究进展[J].中国新药杂志,2007,5(2):158-162.
[13]Venter JC, Adams MD, Myers EW, et al. the sequence of human gonome [J].Science,2001,291:1304-1351.
[14]Wilkins M R, Sanchez J C, Gooley A A, et al. Progress with proteome projects: why all proteins expressed by a genome should be identified and how to do it[J]. Biotech Genet Eng Rev, 1996,13:19-50.
[15]Anderson N L, Anderson N G. Proteome and proteomics: new technologies, new concepts, and new words [J]. Electrophoresis, 1998,19(11):1853-1861.
[16]Service RF.Proteomics.Can Celera Do It Again[J].Science,2000, 287(5461):2136- 2138.
[17]Fenselau C, Yao XD,Reynolds K,et al. Comparative proteomics strategies that exploit mass spectrometry[J]. Abstr Pap Am Chem S, 2001,222:84-90.
[18]Li C, Chen ZC,Xiao ZQ,et al.Comparative proteomics analysis of human lung squam- ous carcinoma[J]. Biochem Bioph Res Co, 2003, 309(1):253-260.
[19]Gygi SP, Corthals GL, Zhang Y,et al. Evaluation of two-dimensional gel electrop- horesis-based proteome analysis technology[J]. Proc NatI Acad SCI U S A,2000, 97(17):9390-9395.
[20]Unlu M, Morgan M E, Minden J S. Difference gel electrophoresis: A single gel method for detecting changes in protein extracts[J]. Electrophoresis, 1997,18(11):2071-2077.
[21]李选文,陈平,梁宋平.癌症差异蛋白质组学研究中样品分离和鉴定分析技术[J].生命科学,2007,19(1):97-102.
[22]Ong S E, Mann M,et al. Mass spectrometry-based proteomic sturns quantitative[J]. NatChemBiol,2005,1(5):252-262
[23]Stulik J,Hernychova L,Prokertova S,et al.Proteome study of colorectal carcinogenesis [J].Eletrophoresis,2001,22:3019-3025.
[24]Celis JE, Gromova I,Moreira JMA,et al.Impact of proteomics on bladder cancer research[J].Phamacogenomics,2004,5(4):381-394.
[25]Celis JE, Korc M. Proteomics of disease[J]. Mol Cell Proteomics, 2005,4(4):345-351.
[26]Takikawa M, Akiyama Y,Maruyama K,et al.Proteomic analysis of a highly metastatic gastric cancer cell line using two-dimensional differential gel electrophoresis[J].Oncol Rep,2006, 16(4):705-711.
[27]Li C,Chen Z,Xiao Z, et al.Comparative proteomeics analysis of human lung squam- ous carcinoma[J].Biochem Biophys Res Commun,2003,309(1):253-260.
[28]Li C,Xiao Z,Chen Z, et al. Proteome analysis of human lung squalors carcinoma [J].proteomics,2006,6(2):547-558.
[29]Yanagiswa K,Shyr Y,Xu B,et al.proteomic pattems of tumour subsets in non-small- cell lung cancer Lancet,2003,362:433-439.
[30]Alfonso P, Catala M, Rico Morales ML, et al. Proteomic analysis of lung biopsies: Differntial protein expression profile between peritumoral and tumoral tissue[J]. Proteomics. 2004,4(2):442-447.
[31]武建国。实用临床免疫学检验[M].江苏科技技术出版社。1990:67-70.
[32]Southern EM. Detection of specific sequences among DNA frag-ments separated by gel electrophoresis. [J] Mol Biol, 1975,98(3):503-17.
[33]Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications[J]. Proc Natl Acad Sci USA, 1979;76(9): 4350-4.
[34]Kurien BT, Scofield RH et al. Multiple immunoblots after non electrophoretic bidirectional transfer of a single SDS-PAGE gel with multiple antigens[J],J Immunol Methods, 1997;205-91.
[35]Hauri HP, Bucher K, Immunoblotting with monoclonal antibody-ties Importance of the blocking solution[J]. Anal Biochem.1986(159):386-389.
[36]Lasne F.Double-blotting: a solution to the problem of nonspecific binding of sencondary antibodies in immunoblotting procedures[J]. J Immunol Methods,2003:276:233.
[37]Lowe G. Slice blotting: a method for detecting the release of immunoreactivesubstances from living brain tissue[J]. j Neurosci Methods, 1990,90:117.
[38]Sumi MG,Mathai A, Sarada C, et al. Rapid diagnosis of tuberculous meningitis by a dot immunobinding assay to detect my cobacterial antigen in cerebrospinal fluid specimens[J]. J Clin Microbiol, 1999,37: 3925.
[39]Szakai D, Schneider G, Pai T. A colony blot immunoassay to identifiy enteroinvasive Escherichia coli and Shigella in stool sample[J]. Diagn Microbiol Infect Dis, 2003,45: 165.
[40]Reese G, Schmechel D, Ayuso R, et al . Grid-immunoblotting: a fast and simple technique to test multiple allergens with small amounts of antibody for cros reactivity[J]. J Chromatogr B Biomed Sci, 2001, 756:151.
[41]Noya O, Alareon de Noya B. The multiple antigen blot assay(MABA): a simple immunoenzymatic technique for simultaneous screening of multiple antigens[J]. IMMUNOL Lett, 1998,63:53.
[42]苏雷,支修益,张毅等.非小细胞肺癌的靶向治疗[J].中华临床医学杂志,2008,2(1):16-18.
[43]Hile M, Freed AL,et al. Possibilities to improve autom ation, speed and precision of proteome anaylysis a comparison of two drmensional electrophoresis and alter natives[J]. Electrophoresis,2001,22:4035-4052.
[44]Hanash S. Disease Proteomics[J]. Nature,2003,422(6928):266-232.
[45]Cohen P. The oriins of protein phosphorylation[J]. Nat cell Biol ,2002,4(5):127-130.
[46]隋少卉,王京兰,钱小红等.磷酸化蛋白质组学分析和定量技术的研究进展.生物化学与生物物理进展,2007,34(3):240-245.
[47]David Y Zhang,Fei Ye,Ling Gao,et al. Proteomics, Pathway array and signaling network based medicine in cancer[J]. Cell Division,2009;1-16.
[48]刘革修.3-磷酸肌醇依赖性蛋白激酶1结构与功能[J].生命科学,2005,17(5):387-391.
[49]Alessi DR, James SR,Downes CP,et al. Characterization of a -3phosphoimositidedependent protein kinase which phosphorylates and activates protein kinase Ba[J].Curr Biol,1997,7:261-269.
[50]Mora A, Komander D,Alessi DR,et al. PDK1, the master regulator of AGC kinase signal transducition[J]. Semin Cell Develop Biol, 2004,15:161-170.
[51]Mauer M, Su T, Saal LH,et al. 3-phosphoinositide-dependent protein kinase-1 potentiates upstream lesions on the phosphatidylinositol 3 kinase patyway in breast carcinoma[J]. Cancer Res,2009,69(15):6299-6306.
[52]Lu Z,Cox-Hipkin MA,WINDSOR WT, Boyapati A,et al. 3-phosphoino- sitide-dependent protein kinase-1 regulates proliferation and survival of cancer cells with an activated mitogen activated protein kinase pathway[J]. Cancer Res, 2010,8(3):421-432.
[53]Dasmahapatra GP, Didolkar P, Alley MC, et al. In vitro combination treatment with perifosine and UCN-01 demonstrates synergism against prostate(PC-3) and lung(A549) epithelial adenocarcinoma cell lines[J]. Clin Cancer Res,2004,10:5442-5252.
[54]Nurse P, Thuriaux P, Nasmyth K,et al. Genetic control of the cell division cycle in the fission yeast schizosaccharomyces pombe[J]. Mol Gen Genet,1976;146(2):167-178.
[55]翟德忠,黄强.细胞周期蛋白依赖性激酶cdc2与恶性肿瘤发生发展的研究.肿瘤,2006,26(5):489-491.
[56]Hunter T, Pines J,et al. Cyclins and cancer:cyclin D and CDK inhibitors come of age[J].cell ,1994,79;573-582.
[57]Kawamoto H, Koizumi H, Uchikoshi T,et al. Expression of the G2-M checkpoint regulators cyclin B1 and cdc2 in non-malignant and malignant human breast lesions immunocyto chemical and quantitative image analyses[J]. Am J pathol,1997,150(1):15-23.
[58]Hansel DE,Dhara S, Huang RC, et al .cdc2/CDK1 expression in esophageal adenoca- rcinoma and precursor lesions serves an a diagnostic and cancer progression marker and potential novel drug target[J]. Am J surg Pathol,2005,29(3):390-399.
[59]Ito Y, Taleda T, Wakasa K,et al. expression of the G2-M modulators in pancreaticadenocarcinoma[J]. Pancreatology, 2002,2(2):138-145.
[60]Kallakury BV, Sheehan CE,Ambros RA,et al.the prognostic significance of p34cdc2 and cyclin D1 protein expression in adenocarcinoma[J].cancer,1997,80(4):753-763.
[61]李国生,张道荣,刘冬等. P34cdc2在非小细胞肺癌中的表达及意义[J].中国医科大学学报,2002,31(2):88-92.
[62]Senju M,Sueoka N,Sato A,et al. Hsp90 inhibitors cause G2/M arrest associated with the reduction of cdc25c and cdc2 in lung cancer cell lines[J]. J cacer Res Clin oncol,2006, 132(3):150-158.
[63]Kyriakis JM,Avruch J,et al. Mammalian mitogen activated protein kinase signal transduction pathways activated by stress and inflammation[J].Physiol Rev,2001,81(2): 807-869.
[64]Brewster JL,De Valori T, Dwyer N,et al. An osmosensing signal trans duction pathway in yeast[J]. Science, 1993,259(5099):1760-1763.
[65]Han J, Lee JD, Bibbs L,et al. A MAP kinase targeted by endotoxin and hyperosmolarity in mammalian cells[J]. Science,1994,265(5173): 808-811.
[66]Obata T, Brown GE, Yaffe MB,et al. MAP kinase pathways activated by stress:the P38MAPK patyway. Crit Care Med,2000,28(4):67-77.
[67]Lee JC, Kumar S,et al. Inhibition of P38 MAP kinase as a therapeutic stategy[J]. Immunopharmacology,2000,47(2):185-201.
[68]张文军,李荣上.P38丝裂原活化蛋白激酶通路及其研究进展[J].医学综述,2007,13(9),663-665.
[69]Ostrander JH, Daniel AR, Lofgren K,et al. Breat tumor kinase(protein tyrosine kinase 6) regulates heregulin induced activation of ERK5 and P-38 MAP kinases in breast cancer cells[J]. Cancer Res, 2007,67(9):4199-4209.
[70]Atsumi T, Kato K, Uno K,et al. Pathophysiological role of the activation of p38 mitogen activated protein kinases in poorly differentiated gastric cancer[J]. Pathol Int, 2007,57(10):635-644.
[71]Junttila MR,Jokilehto T,et al. p38 alpha and p38deltami-togen-activated protein kinase isofroms regulate invasion an growth of head and neck suqamouse carcinoma cells[J]. Oncogene,2007,26(36):5267-5279.
[72]Looda K, Sasaki Y, Horimoto M,et al. Involvement of the p38 mitogen-activated protein kinase cascade in hep-atocellular carcinoma[J]. Cancer, 2003,97(12):3017-3026.
[73]王志强,谢宗涛,陈丽达等.磷酸化p38在非小细胞肺癌中表达的意义[J].肿瘤防治研究,2008,35(6):406-410.
[74]Greenberg AK, Basu S,Hu J, et al. selective p38 activation in human non-small cell lung cancer[J]. Am J Respir Cell Mol Biol,2002,26(5):558-564.
[75]Sturgill TW, Ray BL,et al. Muscle proteins related to microtubule associated protein-2 and sub-strates for and insulin-stimulatable kinase[J]. Biochem Biophys Res Commun, 1986,134(2):565-571.
[76]Boulton TG, Yancopoulos GD, Gregory JS,et al. An insulin-stimu-lated protein kinade similar to yeast kinases involved in cell cycle contro[J]. Sciences,1990,249(4964):64-67.
[77]Gary L.Johnson, Razvan Lapadat,et al. The protein kinase complement of the human genome[J]. Science,2002;296(6):1911-1912.
[78]Robinson MJ, Cobb MH,et al.Mitogen activated protein kinase pathways[J]. Curr Opin Cell Biol,1997,9(2):180-186.
[79]Ito Y, Sasaki Y, Horimoto M,et al. Activation of mitogen-activated protein kinases/extracellular signal-regula ted kinases in human hepatocellular carcinoma[J]. Hepato- logy,1998,27(4):951-958.
[80]Adeyinka A, Kui Y,Cherlet T,et al. Activated mitogen-activated protein kinase expression during human breast tumorigenesis and breast cancer progression[J]. Clin Cancer Res,2002,8(6):1747-1753.
[81]谷艳娇,包翠芬,高志安等.ERK1/2和p-ERK1/2在肺癌组织中的表达及意义[J].中国组织化学与细胞化学杂志.2009.18(3):328-331.
[82]Blackhall FH, Pintilie M, Michael M, et al. Expression and prognostic significance of kit, Protein kinse B,and mitogen-activated protein kinase in patients with small cell lung cancer[J]. Clin Cancer RES, 2003,9(6):2241-2247.
[83]Montrminy MR,sevarino KA,Wagner JA, et al.Identification of a cyclic AMP responsive element within the rat somatostatin gene[J].Proc Natl Acad Sic USA,1986, 83:6682-6686.
[84]徐瑞元,王燕峰,徐长发. CREB研究进展[J].中国生物工程学. 2003,23(1):39-47.
[85]Kathleen M. Sakamoto, David A. Frank. CREB in the pathophysiology of Cancer: Implications for Targeting Transcription Factors for Cancer Therapy[J]. Clin Cancer Res 2009,15(8):2583-2587.
[86]Kathleen M, Sakamoto, David A, et al. CREB in the Pathophysiology of cancer:lim- plications for targetin transcription factors for cancer therapy[J]. Clin cancer Res,2009,15(8): 2583-2587.
[87]Sofi M, Young MJ, Papamakarios T, et al. Role of CRE-binding protein (CREB) in aromatase expression in breast adipose[J]. Breast Cancer Res Treat 2003,79: 399–407.
[88]Unni E, Sun S, Nan B, et al. Changes in androgen receptor nongenotropic signaling correlate with transition of LNCaP cells to androgen independence[J]. Cancer Res 2004,64: 7156–68.
[89]Crans-Vargas HN, Landaw EM, Bhatia S, et al. Expression of cyclic adeno-sine monophosphate response-element binding protein in acute leukemia[J]. Blood 2002,99: 2617–2619.
[90]Hye-sook seo,Dianei D Liu, Ja seok koo.et al. Cyclic AMP Response Element- Binding Protein Over expression : A Feature Associated with Negative prognosis in Never Smokers with NSCLC[J]. Cancer Res 2008,68(15):6065-6073.
[91]Sita Aggarwal, Seung-Wook Kim, Ja Seok Koo, et al. Growth Suppression of LungCancer Cells by Targeting Cyclic AMP Response Element-Binding Protein[J]. Cancer Res 2008:68:(4),981-988.
[92]Seaki K, You A, Takaku F,et al. cell-cycle-regulated phosphorylation of cAMP response element-bunding protein:identification of nover phosphorylation sites[J].Biochem J,1999,338(1):49-54.
[93]Sutherland EW,Robison GA.,et al. The role of cyclic-3`5`-AMP in responses to catecholamines and other hormones[J]. Pharmaco Rev,1966,18(1):145-161.
[94]Montrminy MR,Bilezikjan LM. Binding of a nuclear protein to the cyclic AMP response element of the somatostatin gene[J]. Nature, 1987,328:175-178.
[95]Yamamoto KK,Gonzalez GA,Biggsl WH,et al.Phosphorylation induced binding and transcriptional efficacy of factor CREB[J].Nature,1988,334:494-498.
[96]Gonzalez GA,Yamamoto kk,Fischer WH,et al. A cluster of phosphorylation sites on the cyclic AMP regulated nuclear factor CREB predicted by its sequence[J]. Nature, 1989, 337:749-752.
[97]Berkowitz LA, Gliman MZ, et al. Two distinct forms of active transcription factor CREB(c AMP response element binding protein)[J],Proc Natil Acad Sci U S A,1990, 87(14):5258-5262.
[98]Ruppert S,Cole TJ, Boshart M, Schmid E,schutz G. Multiple mRNA isoforms of the transcription activator protein CREB:generation by altemative splicing and specific expression in primary spermatocytes[J]. EMBO J,1992,11(4):1503-1512.
[99]Gonzalez GA, Montminy MR.Cyclic cAMP stimulates somatostatin gene transcription by phosphorylaton of CREB at ser 133[J]. Cell 1989,59(4):675-680.
[100]Lee CQ, Yun Y, Hoeffler JP, et al. Cyclic AMP responsive transcription activation of CREB-327 in volves interdependent phosphorylated subdomains[J]. EMBOJ ,1990,9(13): 4445-4465.
[101]Kevin AW, Noma M ,et al. Transcription regulation by CREB and its relatives[J].Biochim Biophys Acta,1993,1174:221-233.
[102]Craig JC,Schumacher MA,Mansoor SE ,et al.Consensus and variant cAMP- regulated enhancers have distinct CREB-binding properties[J].J Biol Chem.2001, 276(15): 11719-28.
[103] Ribeiro A, Brown A, Lee KA. An in vino assay for members of the Camp response element binding protein family of transcription factors[J].1994,269(49):31124-31128.
[104]窦爱霞,董建华.cAMP信号途径和基因表达调控[J].上海第二医科大学学报,2004,24(12):1070-1073.
[105]Saeki K, You A et al. Distinct involvement of cAMP response element dependent transcriptions in functional and morphological maturation during retinoid mediated human myeloid differentiation[J]. Journal of leukocyte biology,2003,73(5):673-681.
[106]Sudzinski GP, Harrison LE, et al. Differentiation-related changes in the cell cycle traverse[J]. Int Rev Cytol, 1999,189:1-58.
[107]Lee HJ,Mignacca RC, sakamoto KM. Transcriptional activation of egr-1 by granulocyte macrophage colony stimulating factor but not interleukin 3 requires phosphory- lation of cAMP response element binding protein(CREB) on serine 133[J]. J Biol Chem,1995,270(27):15979-15983.
[108]Haywitz A, Greenbeng M. CREB: a stimulus induced transcription factor activated by a diverse array of extracellular signals[J]. Annual Review of Biochemistry, 1999,68: 821-861.
[109]Gonzalez GA, yamamoto KK, Fischer WH, et al. A cluster of phosphorylation sites on the cyclic AMP-regulated nuclear factor CREB predicted by its sequence[J]. Nature 1989;337:749-752.
[111]Swanson DJ, Zellmer E, Elaine JL. The homeodomain protein Arix interacts synergistically With cyclic AMP to regulate expression of neurotransmitter biosynthetic genes[J]. J Biol Chem,1997,272(43): 27382-27392.
[112]Matthews RP, Guthrie CR, Wailes LM et al. Calcium/calmodulin-depen-dent protein kinase typesⅡandⅣdifferentially regulate CREB-dependent gene expression[J]. Mol cell boil 1994;14:6107-6116J.
[113]Sun P, Enslen H, Myung PS, Maurer RA. Differential activation of CREB by Ca2+/ calmodulin-depen-dent protein kinase typeⅡand typeⅣinvolves phosphor-ylation of a site that negatively regulates activity[J]. Genes Dev 1994;8:2527-2539.
[114]Heist EK, Srinvasan M, Schulman H. Phosphorylation at the nuclear localization signal of Ca2+/calmodulin dependent protein kinaseⅡblocks its nuclear targeting[J]. J Bilo Chem, 1998,273(31):19763-19771.
[115]Du K, Montminy M. CREB is a regulatory target for the protein kinase AKT/PKB[J]. J Bilo Chem 1998,273:32377-32379.
[116]Philipp V,Carsten B, Bernd Nu,et al. Assigning Funcional Domains within the P101 Regulatory subunit of phosphoinositide 3 kinase[J]. J Biol Chem,2005,280:5121-5127.
[117]Murthy SS, Tosolini A,Taguchi T, et al. Mapping of AKT, encoding a member of the Akt/Protein Kinase M family, to human and rodent chromosomes by fluorescence in situ hybridization[J] . Cytogenet Cell Genet,2000,88:38-40.
[118]Downward J. Mechanisms and consequences of activation of protein kinaseB/AKT [J].Curr Opin Cell Biol,1998,10(2):262-267.
[119]Chepurny OG, Hussain MA, Holz GG,et al. Exendi-4 as a stimulator of rat insulin I gene promoter activity via bZIP/CRE interactions sensitive to serine/threonine protein kinase inhibitor RO 31-8220[J]. Endocrinology 2002; 143;2303-2313.
[120]Tan Y, Rouse J, Zhang A comb MJ,et al.Fgf and stress regulate CREB and ATF-1 via a pathway involing P38 MAP kinase and MAPKAP kinase-2[J]. EMBO J 1996;15:4629-4642.
[121]Force T, Bonventre JV,et al. Growh factors and mitogen activated protein kinases[J]. Hypertension,1998,21:152-161.
[122]Roux pp, Richards SA,Blenis J,et al. Phosphorylation of P90 ribosomal S6 kinase(RSK) regulates extracellular signal regulated kinase docking and RSK activity[J]. Mol Cell Biol,2003,23(14):4796-4804.
[123]Scheid MP, Foltz IN, Young PR, Schrader JW et al. Ceramide and cyclic adenosine monophosphate(cAMP) induce cAMP response element binding protein phosphorylation via distinct signaling pathways while having opposite effects on myeloid cell surviva[J]. Blood 1999; 93:217-225.
[124]Yamamoto KK, Gonzalez GA, Montminy MR, et al. phosphorylation induced binding and transcriptional efficacy of nuclear factor CREB[J]. Nature 1988, 334:484-498.
[125]Zanassi p, paolillo M,feliciello A et al. cAMP dependent protein kinase induces cAMP response element binding protein phosphorylation via an intracellular calcium release/ERK dependent pathway in striatal neurons[J]. J boil chem,2001,276(15):11487-11495.
[126]Mayr BM, Cancetteri G, Montminy MR,et al. Distinct effects of cAMP and mitogenic signals on CREB-binding protein recruitment impart specificity to target gene activa-tion via CREB[J]. Proc Natl Acad Sci USA,2001,98:10936-10941.
[127]Kee BL, Arias J, Montminy MR,et al. Adaptor-mediated recruitment of RNA polymeraseⅡto a signal-dependent activator[J]. J boil Chem 1996,271(5):2373-2375.
[128]Nakajima T, Uchida C, Anderson SF,Montminy MR,et al. Analysis of a cAMP responsive activator reveals a two-component mechanism for transcriptional induction via signal dependent factors[J]. Genes Dev,1997,11(6):738-747.
[129]Schiliz RL, Mizzen CA, Vassilev A, et al. Overlapping but distinct pattems of histone acetylation by the human coactivators p300 and PGAF with in nucleosomal substrates[J]. J Biol CHEM, 1999,274(3):1189-1192.
[130]Kathleen M, Sakamoto, David A, et al. CREB in the Pathophysiology of cancer:limplications for targetin transcription factors for cancer therapy[J]. Clin cancer Res,2009, 15(8): 2583-2587.
[131]Montarolo PG, Kandel ER, Schacher S. Long term heterosynaptic inhibition in(RSK) regulates extracellular signal regulated kinase docking and RSK activity[J]. Mol Cell Biol,2003,23(14):4796-4804.
[123]Scheid MP, Foltz IN, Young PR, Schrader JW et al. Ceramide and cyclic adenosine monophosphate(cAMP) induce cAMP response element binding protein phosphorylation via distinct signaling pathways while having opposite effects on myeloid cell surviva[J]. Blood 1999; 93:217-225.
[124]Yamamoto KK, Gonzalez GA, Montminy MR, et al. phosphorylation induced binding and transcriptional efficacy of nuclear factor CREB[J]. Nature 1988, 334:484-498.
[125]Zanassi p, paolillo M,feliciello A et al. cAMP dependent protein kinase induces cAMP response element binding protein phosphorylation via an intracellular calcium release/ERK dependent pathway in striatal neurons[J]. J boil chem,2001,276(15):11487-11495.
[126]Mayr BM, Cancetteri G, Montminy MR,et al. Distinct effects of cAMP and mitogenic signals on CREB-binding protein recruitment impart specificity to target gene activa-tion via CREB[J]. Proc Natl Acad Sci USA,2001,98:10936-10941.
[127]Kee BL, Arias J, Montminy MR,et al. Adaptor-mediated recruitment of RNA polymeraseⅡto a signal-dependent activator[J]. J boil Chem 1996,271(5):2373-2375.
[128]Nakajima T, Uchida C, Anderson SF,Montminy MR,et al. Analysis of a cAMP responsive activator reveals a two-component mechanism for transcriptional induction via signal dependent factors[J]. Genes Dev,1997,11(6):738-747.
[129]Schiliz RL, Mizzen CA, Vassilev A, et al. Overlapping but distinct pattems of histone acetylation by the human coactivators p300 and PGAF with in nucleosomal substrates[J]. J Biol CHEM, 1999,274(3):1189-1192.
[130]Kathleen M, Sakamoto, David A, et al. CREB in the Pathophysiology of cancer:limplications for targetin transcription factors for cancer therapy[J]. Clin cancer Res,2009, 15(8): 2583-2587.
[131]Montarolo PG, Kandel ER, Schacher S. Long term heterosynaptic inhibition in
[142]Unni E, Sun S, Nan B, et al. Changes in androgen receptor nongenotropic signaling correlate with transition of LNCaP cells to androgen independence[J]. Cancer Res 2004, 64:7156-7168.
[143]Crans vargas NH, Landaw EM, Bhatia S,et al.Expression of cyclic adeno-sime monophosphate response-element binding protein in acute leukemia[J]. Blood 2002, 99: 2617-2619.
[144]Shankar DB,Cheng JC, Kinjo K,et al. the role ofCREB as a proto-oncogene in hematopoiesis and in acute myeloid leukemia[J]. Cancer Cell.2005,7:351-362.
[145]Linnerth NM, Baldwin M, Campbell C,Moorehead RA,et al. IGF-Ⅱinduces CREB phosphorylation and cell survival in human lung cancer cells[J]. Oncogene 2005,24: 7310-1319.
[146]Hye-Sook Seo, Diane D Liu, Ja Seok Koo, et al. Cyclic AMP response element binding protein overexpression: A Feature associated with negative prognosis in never smokers with non small cell lung cancer[J].Cancer Res 2008,68(15):6065-6073.
[147]Kononen J,Bubendorf L,Kallioniemi A, et al.Tissue microarrays for high- throughput molecular profiling of tumor specimens[J].Nature 1998,4:844.
[148]王新允,朱丛中,刘婷等.组织芯片研制的几点体会[J].天津医科大学学报,2005,11(1),24-25.
[149]周小鸽,王鹏,刘金香等.组织芯片的基本应用范围.诊断病例学杂志;2003,10(6),350-352.
[150]Parkin DM,Muir CS,Whelan SL,et al.Cancer Incidence in Five Continents,vols,ⅠtoⅧ[R].Lyon:IARC, 2005.
[151]Lonescu AM, Schwarz EM, Vinson C, et al. PTHrP modulates chondrocyte differentiation through AP-1 and CREB signaling[J]. J Biol Chem 2001,276:11639-11647.
[152]Conkright MD, Montminy M. CREB:the unindicted cancer co-conspirator[J]. Trends Cell Biol 2005,15:457-459.
[153]Shankar DB,S akamoto KM.the role of cyclic-AMP binding protein(CREB) in leukemia cell prolife ration and acute leukemias[J]. Leuk Lymphoma 2004, 45:265-270.
[154]Oike Y, Takakura N,Hata A,et al.Mice homozygous for a truncated form of CREB binding protein exhibit defects in hematopoiesis and vasculo-angiogenesis[J]. Blood 1999,93:2771-2779.
[155]Stylianou E, Saklatvala J. Interleukin-1[J]. Int J Biochem cell boil,1998,30: 1075-1079.
[156]Nyormoi O, Bar-Eli M. Transcriptional regulation of metastasis-related genes in human melanoma[J]. Clin Exp Metastasis 2003,20:2 51-263.
[157]Annan RS, Huddleston MJ,verna R,et al. A multidimensional Electrospray MS-Based Approach to phosphopeptide Mapping[J]. Anal Chem,2001, 73(3):393-404.
[158]Bodenmiler B, Mueller LN, Mueller M,et al. Reproducible isolation of distinct overlapping segments of the posphoproteome[J]. Nat Methods.2007,4(3):231-237.
[159]Sita Aggarwal, Seung Wook Kim, Ja Seok Koo, et al. Growth suppression of lung cancer cells by targeting Cyclic AMP Response Element Binding Protein[J]. Cancer Res 2008, 68(4),981-988.
[160]Sharma H,Sen S, Mathur M,et al.Combined evaluation of expression of telomerase,surviving,and anti apoptotic BCL-2 family members in relation to loss of differentiation and apoptosis in human bead and neck cancers[J]. Head Neck, 2004,26 (8):733-740.
[161]Stepulak A,sifringer M,Rzeski W,et al.AMPA antagonists inhibit the extracellular signal regulated kinase athway and suppress lung cancer growth[J]. Cancer Biol Ther 2007,6:1908-1915.
[162]Shouwei Han, Neil Sidell, Susanne Roser Page,et al. Fibronectin stimulates human lung cancer carcinoma cell growth by inducing cyclooxygenase-2(cox-2) expression[J].Cancer, 2004;111:322-331.
[163]Au JS, Cho WC,Law SC,et al. Protemic approach to biomarker discovery in cancer tissue from lung adenocarcinoma among nonsmoking Chinese women in Hong Kong[J].Cancer Invest 2008, 26:128-135.
[164]Hildegrad M, Maria Cekanova,et al.NNK-induced hamster lung adenocarcinomas over expressβ-adrenergic and EGFR signaling pathways[J].lung cnacer,2005,49:35-45.
[165]Goldkon T,Khan E,et al. EGF receptor expose to cigarette smoke undergoes aberrant activation and trafficking. American Association for Cancer Ressearch Annual Meeting: Proceedings, 2009,Denver CO.phila-delphia,pa:AACR 2009.
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