扶正增效方对肺癌放射增敏的分子机制研究
摘要
原发性肺癌目前是全球发病率与死亡率较高的恶性肿瘤之一,近年来我国肺癌发病率与死亡率呈逐年上升趋势,现已成为我国城市人口癌症死亡的首要原因。原发性肺癌患者约70%需接受放射治疗,但由于癌组织中乏氧细胞的大量存在、抗放射损伤等,在一定程度上限制了放疗的应用,放射增敏剂不仅使肿瘤组织对放射线的敏感性大大增加,局部控制程度明显提高,同时还可减少肿瘤局部复发和转移,因此对肺癌放射增敏剂的研究具有重要深远的意义。
RNAi是一种与靶基因同源的双链RNA诱导的特异转录后基因沉默现象,具有高效、高特异性及对靶基因高选择性等特点,能高效抑制特定基因的表达,效果接近于基因敲除技术,目前已广泛应用于各种肿瘤的发病机制及肿瘤基因治疗研究中。
从前期的结果中我们认为扶正增效方具备成为放射增敏剂的基本条件,其放射增敏作用的主要蛋白是S100A9、CyPA,但这只是定性的初步研究、而不是定量的研究,而且具体哪种基因发挥主要作用尚不清楚,本课题为国家自然科学基金(NO.81072925)资助课题,用q-PCR、Western-blot等技术和RNAi等方法验证和从量上分析了中药对这两个基因的放射增敏作用。
目的：具体S100A9、亲环素A在扶正增效方对肺癌放射增敏中的作用和地位。用RNAi技术验证放射增敏相关靶点—S100A9、CyPA,此将为确定放射增敏相关预测因子和放射增敏药物的研发具有重要意义。
方法：建立裸鼠人肺腺癌移植瘤模型,分4组,空白组、中药组、放射组、中药+放射组,当瘤积达1cm3左右时,一次给予放射组、中药+放射组瘤体10Gy剂量照射。观察抑瘤率；Western blot检测S100A9、CyPA蛋白表达；用实时荧光定量RT-PCR法检测S100A9、CyPA mRNA表达；慢病毒介导目的基因敲降技术沉默肺腺癌S100A9、CyPA, Western blot筛选出S100A9和CyPA RNAi有效的干扰靶点序列。干扰后分为空白组、阴性对照组、RNAi S100A9组、RNAiCyPA组,照射后通过集落形成实验计算克隆生成率和存活分数,利用单击多靶模型评价RNAi技术沉默肺腺癌S100A9、CyPA后细胞辐射敏感性,流式细胞仪测定细胞周期。
结果：通过实验发现扶正增效方有一定抑瘤作用,与对照组、中药组、放射组比较,中药加放射组抑瘤率显著增高。Western blot结果显示：放射后6h,与对照组比较,S100A9、CyPA蛋白各组表达有所下降,但无差异。放射后12h,与对照组比较,中药+放射组蛋白表达降低,有显著性差异(P<0.05)。放射后24h,S100A9蛋白表达,放射组与对照组比较降低,有显著性差异(P<0.05)。中药+放射组表达相对各组均有显著性差异(P<0.05)。CyPA蛋白表达,放射组与对照组、中药组比较均降低,有显著性差异(P<0.05)。中药+放射组表达相对各组均降低,且有显著性差异(P<0.05)。实时荧光定量PCR结果显示：放射后6h,与对照组比较,各组S100A9、CyPA mRNA各组表达有所下降,但无差异。放射后12h,与对照组比较,放射组S100A9mRNA表达下降,有差异(P<0.05)。中药+放射组与对照组、中药组比较均降低,有显著性差异(P<0.05)。放射后12h,与对照组比较,放射组、中药+放射组CyPA mRNA表达下降,有显著性差异(P<0.05)。放射后24h, S100A9、CyPA mRNA表达,放射组与对照组、中药组比较均降低,有显著性差异(P<0.05)。中药+放射组表达相对各组均有下降,且有显著性差异(P<0.05)。进一步构建S100A9RNAi和CyPA RNAi'慢病毒表达质粒并包装出慢病毒,通过Westernblot筛选出S100A9和CyPA RNAi有效的干扰靶点序列,制备了S100A9和CyPA shRNA/PAa稳转细胞。通过集落形成实验计算克隆生成率和存活分数,利用单击多靶模型得出RNAi (S100A9、CyPA)后降低了肿瘤细胞SF2Gy值、Do值、Dq值,增加了辐射效应,增强射线对肿瘤细胞的杀伤作用。流式细胞仪检测发现,RNAi沉默S100A9和CyPA后,未照射时,各组G0/G1、S的细胞有差异,但无统计学意义,G2/M期细胞,S100A9组相对空白组增加,有统计学意义(P<0.05),CyPA组相对空白组和对照组增加,有统计学意义(P<0.05)。2Gy照射后,S100A9和CyPA组相对空白组和对照组,Go/G1期细胞明显减少(P<0.05),G2/M期细胞显著增多(P<0.05),S100A9组S期细胞降低,但无差异,CyPA组S期细胞显著降低(P<0.05)。
结论：本实验证明了扶正增效方对肺腺癌具有一定的抑瘤作用和较强的放射增敏作用。通过Western blot和q-PCR检测S100A9、CyPA表达,发现扶正增效方通过下调S100A9和CyPA表达对肺腺癌起到放射增敏作用。尤其在放射后24小时表达显著降低,差异有统计学意义。进一步采用慢病毒介导的S100A9和CyPA基因沉默,Westerblot筛选出有效的干扰靶点序列,制备了S100A9和CyPA shRNA/PAa稳转细胞。在体外实验发现慢病毒介导的S100A9和CyPA shRNA能使细胞阻滞在G2/M期的比例增高,细胞辐射敏感性增高。
Currently,Lung cancer is one of the high morbidity and mortality malignant tumor worldwide, and the incidence and mortality of lung cancer are increasing year by year.It has become the leading cause of cancer death in urban population in China. About70%patients of primary lung cancer need to be accepted radiotherapy, but the hypoxic cell carcinoma and resistance to radiation damage, which it partly limit the use of radiotherapy.Not only radiation sensitizer improve tumor sensitivity to radiation and the level of local control greatly, but also it can reduce recurrence and metastasis of the local cancer, so studying of radiosensitizer have an important and far-reaching significance.
RNA interference is a gene silencing phenomenon after specific transcription,which induced by double-strand RNA of homologous to the target gene.It has the advantages characteristics of high efficiency, high specificity and high selectivity to the target gene,could effectively inhibit expression of specific gene, and the effect is close to the gene knockout technology,which has been widely used in the study on the pathogenesis and tumor gene therapy.
From the previous results,we concluded that Fuzheng Zengxiao formula has the basic conditions of radiosensitizer, the main protein of the radiation sensitization is S100A9, CyPA, However, this is only a preliminary qualitative study, rather than quantitative research,Which specific genes play a major role is unclear,which it is need validation and analysis radiation sensitization of traditional Chinese medicine to two genes in q-PCR, Western-blot technology and RNA interference.
Objective:To evaluate the role and status of S100A9, and CyPA in Fuzheng Zengxiao formula increasing radiotherapy sensitivity of lung cancer.The present project was supported by National Natural Science Foundation of China (NO.81072925),Using RNAi technology to verify the radiosensitizing target-S100A9and CyPA, which it is importance to determine predictive factors of radiosensitivity and research of radiosensitizing drugs.
Method:Establishment the model of transplanted human lung adenocarcinoma in nude mice, then divided into4groups, including blank group, Chinese medicine group, radiation group, Chinese medicine+radiation group, when volume of the tumor reach1cm3, we will give10Gy dose irradiation for radiation group and Chinese medicine+radiation group.Observe tumor inhibition rate;identification of protein expression by Western.Detection mRNA expression of S100A9and CyPA by real-time fluorescent quantitative RT-PCR. The silence of S100A9and CyPA in lung adenocarcinoma by gene knockdown Lentivirus-mediat ed,selecting effective interference sequence target of S100A9and CyPA by Western.Itis divided into blank group, negative control group, RNAi S100A9group, and RNAi CyPA group after Interference,then calculating the clone forming rate and survival fraction by colony forming experiment after irradiation,Evaluating cell radiation sensitivity of RNAi silencing the S100A9, CyPA in lung cancer by Click multiple target model, and cell cycle analyzed by flow cytometry.
Result:Fuzheng Zengxiao formula has antitumor effects,which compared with the control group, Chinese medicine group,and radiation group,Tumor inhibition rate of traditional Chinese medicine+radiation group was striking increased.Results of Western blot showed:at6h after radiotherapy, that compared with control group, protein expression of S100A9and CyPA were decreased in each group, but no statistical difference.at12h after radiotherapy, Compared with the control group,expression of Chinese medicine+radiation group were decreased, and there were statistical significant difference(P<0.05).at24h after radiotherapy,The protein expression of S100A9,compared with the control group,radiation group was decreased,and there were statistical significant difference(P<0.05).To the other groups,expression of Chinese medicine+radiation group are decreased,there are significant.Co mpared with the control group,the protein expression of CyPA in the Chinese medicine group and radiation group was decreased,and there were statistical significant differences(P<0.05).T o the other groups,The protein expression of CyPA in Chinese medicine+radiation group were decreased,and there were statistical significant difference.Results of Real-time quantitative PCR show:at6h after radiotherapy,compared with control group,mRNA expression of S100A9and CyPA were decreased in the other group, but there were no difference. At12h after radiotherapy,Compared with the control group,mRNA expression of S100A9of radiation group were decreased,and there were statistical significant difference(P<0.05).Compared with the control group and Chinese medicine,Chinese medicine+radiation group were decreased,th ere are statistical significant difference (P<0.05).mRNA expression of CyPA,Compared with the control group,radiation group and Chinese medicine+radiation group are decreased,there are statistical significant difference(P<0.05).at24h after radiotherapy, mRNA expression of S100A9and CyPA, Compared with the control group、Chinese medicine,radiation group are decreased,there are statistical significant difference(P<0.05).To the other groups,expression of Chinese medicine+radiation group were decreased,there were statistical significant difference (P<0.05).Furthe, Construction of the lentiviral expression plasmid S100A9and CyPA RNAi gene and packing lentiviral.Select effective interference target sequence of S100A9and CyPA RNAi by Western blot,and stably transfected cells of S100A9and CyPA shRNA/PAa were prepared.It calculated the clone forming rate and survival fraction by the colony forming experiment,Using click multiple target model,it find that reduce SF2Gy value, Do value, Dq value of tumor cell after RNAi (S100A9,cyclophilin A),Increasing the radiation effect and enhance cytotoxic effects of radiation on tumour cells.The results of flow cytometry,after RNAi silencing S100A9and CyPA,without radiotherapy,Go/G1,S phase incells of every Groups had difference,but there were no statistical difference.G2/M phase in cells, Compared with the blank group, the S100A9group increased,there were statistical significant difference(P<0.05).Compared with the blank group and control group,CyPA group were increased,there were significant difference(P<0.05).After2Gy radiotherapy,Compared with the blank group, control group, G0/G1cells of S100A9and CyPA group were decreased,G2/M cells are statistical significanly increased(P<0.05).S cells of S100A9group were decreased,but no statistical difference.S cells of CyPA group were significantly decreased(P<0.05).
Conclusion:The experiment demonstrated that Fuzheng Zengxiao formula had certein damps the lump rate and better radiosensitizing effect on lung adenocarcinoma.Detection the expression of S100A9and CyPA by Western blot and q-PCR,it discoved that Fuzheng Zengxiao formula played radiosensitizing effect by down-regulating expression of S100A9and CyPA in lung adenocarcinoma.24h after radiotherapy,the express was significantly decreased,and there were statistical significant difference(P<0.05).Further,Construction of the lentiviral expression plasmid S100A9and CyPA RNAi gene,select effective interference target sequence by Western blot, and stably transfected cells of S100A9and CyPA shRNA/PAa were prepared.In vitro experiment,S100A9and CyPA siRNA mediated Lentivirus can improve ratio of G2/M phase in cell,and increase radiation sensitivity of cell.
RNAi是一种与靶基因同源的双链RNA诱导的特异转录后基因沉默现象,具有高效、高特异性及对靶基因高选择性等特点,能高效抑制特定基因的表达,效果接近于基因敲除技术,目前已广泛应用于各种肿瘤的发病机制及肿瘤基因治疗研究中。
从前期的结果中我们认为扶正增效方具备成为放射增敏剂的基本条件,其放射增敏作用的主要蛋白是S100A9、CyPA,但这只是定性的初步研究、而不是定量的研究,而且具体哪种基因发挥主要作用尚不清楚,本课题为国家自然科学基金(NO.81072925)资助课题,用q-PCR、Western-blot等技术和RNAi等方法验证和从量上分析了中药对这两个基因的放射增敏作用。
目的：具体S100A9、亲环素A在扶正增效方对肺癌放射增敏中的作用和地位。用RNAi技术验证放射增敏相关靶点—S100A9、CyPA,此将为确定放射增敏相关预测因子和放射增敏药物的研发具有重要意义。
方法：建立裸鼠人肺腺癌移植瘤模型,分4组,空白组、中药组、放射组、中药+放射组,当瘤积达1cm3左右时,一次给予放射组、中药+放射组瘤体10Gy剂量照射。观察抑瘤率；Western blot检测S100A9、CyPA蛋白表达；用实时荧光定量RT-PCR法检测S100A9、CyPA mRNA表达；慢病毒介导目的基因敲降技术沉默肺腺癌S100A9、CyPA, Western blot筛选出S100A9和CyPA RNAi有效的干扰靶点序列。干扰后分为空白组、阴性对照组、RNAi S100A9组、RNAiCyPA组,照射后通过集落形成实验计算克隆生成率和存活分数,利用单击多靶模型评价RNAi技术沉默肺腺癌S100A9、CyPA后细胞辐射敏感性,流式细胞仪测定细胞周期。
结果：通过实验发现扶正增效方有一定抑瘤作用,与对照组、中药组、放射组比较,中药加放射组抑瘤率显著增高。Western blot结果显示：放射后6h,与对照组比较,S100A9、CyPA蛋白各组表达有所下降,但无差异。放射后12h,与对照组比较,中药+放射组蛋白表达降低,有显著性差异(P<0.05)。放射后24h,S100A9蛋白表达,放射组与对照组比较降低,有显著性差异(P<0.05)。中药+放射组表达相对各组均有显著性差异(P<0.05)。CyPA蛋白表达,放射组与对照组、中药组比较均降低,有显著性差异(P<0.05)。中药+放射组表达相对各组均降低,且有显著性差异(P<0.05)。实时荧光定量PCR结果显示：放射后6h,与对照组比较,各组S100A9、CyPA mRNA各组表达有所下降,但无差异。放射后12h,与对照组比较,放射组S100A9mRNA表达下降,有差异(P<0.05)。中药+放射组与对照组、中药组比较均降低,有显著性差异(P<0.05)。放射后12h,与对照组比较,放射组、中药+放射组CyPA mRNA表达下降,有显著性差异(P<0.05)。放射后24h, S100A9、CyPA mRNA表达,放射组与对照组、中药组比较均降低,有显著性差异(P<0.05)。中药+放射组表达相对各组均有下降,且有显著性差异(P<0.05)。进一步构建S100A9RNAi和CyPA RNAi'慢病毒表达质粒并包装出慢病毒,通过Westernblot筛选出S100A9和CyPA RNAi有效的干扰靶点序列,制备了S100A9和CyPA shRNA/PAa稳转细胞。通过集落形成实验计算克隆生成率和存活分数,利用单击多靶模型得出RNAi (S100A9、CyPA)后降低了肿瘤细胞SF2Gy值、Do值、Dq值,增加了辐射效应,增强射线对肿瘤细胞的杀伤作用。流式细胞仪检测发现,RNAi沉默S100A9和CyPA后,未照射时,各组G0/G1、S的细胞有差异,但无统计学意义,G2/M期细胞,S100A9组相对空白组增加,有统计学意义(P<0.05),CyPA组相对空白组和对照组增加,有统计学意义(P<0.05)。2Gy照射后,S100A9和CyPA组相对空白组和对照组,Go/G1期细胞明显减少(P<0.05),G2/M期细胞显著增多(P<0.05),S100A9组S期细胞降低,但无差异,CyPA组S期细胞显著降低(P<0.05)。
结论：本实验证明了扶正增效方对肺腺癌具有一定的抑瘤作用和较强的放射增敏作用。通过Western blot和q-PCR检测S100A9、CyPA表达,发现扶正增效方通过下调S100A9和CyPA表达对肺腺癌起到放射增敏作用。尤其在放射后24小时表达显著降低,差异有统计学意义。进一步采用慢病毒介导的S100A9和CyPA基因沉默,Westerblot筛选出有效的干扰靶点序列,制备了S100A9和CyPA shRNA/PAa稳转细胞。在体外实验发现慢病毒介导的S100A9和CyPA shRNA能使细胞阻滞在G2/M期的比例增高,细胞辐射敏感性增高。
Currently,Lung cancer is one of the high morbidity and mortality malignant tumor worldwide, and the incidence and mortality of lung cancer are increasing year by year.It has become the leading cause of cancer death in urban population in China. About70%patients of primary lung cancer need to be accepted radiotherapy, but the hypoxic cell carcinoma and resistance to radiation damage, which it partly limit the use of radiotherapy.Not only radiation sensitizer improve tumor sensitivity to radiation and the level of local control greatly, but also it can reduce recurrence and metastasis of the local cancer, so studying of radiosensitizer have an important and far-reaching significance.
RNA interference is a gene silencing phenomenon after specific transcription,which induced by double-strand RNA of homologous to the target gene.It has the advantages characteristics of high efficiency, high specificity and high selectivity to the target gene,could effectively inhibit expression of specific gene, and the effect is close to the gene knockout technology,which has been widely used in the study on the pathogenesis and tumor gene therapy.
From the previous results,we concluded that Fuzheng Zengxiao formula has the basic conditions of radiosensitizer, the main protein of the radiation sensitization is S100A9, CyPA, However, this is only a preliminary qualitative study, rather than quantitative research,Which specific genes play a major role is unclear,which it is need validation and analysis radiation sensitization of traditional Chinese medicine to two genes in q-PCR, Western-blot technology and RNA interference.
Objective:To evaluate the role and status of S100A9, and CyPA in Fuzheng Zengxiao formula increasing radiotherapy sensitivity of lung cancer.The present project was supported by National Natural Science Foundation of China (NO.81072925),Using RNAi technology to verify the radiosensitizing target-S100A9and CyPA, which it is importance to determine predictive factors of radiosensitivity and research of radiosensitizing drugs.
Method:Establishment the model of transplanted human lung adenocarcinoma in nude mice, then divided into4groups, including blank group, Chinese medicine group, radiation group, Chinese medicine+radiation group, when volume of the tumor reach1cm3, we will give10Gy dose irradiation for radiation group and Chinese medicine+radiation group.Observe tumor inhibition rate;identification of protein expression by Western.Detection mRNA expression of S100A9and CyPA by real-time fluorescent quantitative RT-PCR. The silence of S100A9and CyPA in lung adenocarcinoma by gene knockdown Lentivirus-mediat ed,selecting effective interference sequence target of S100A9and CyPA by Western.Itis divided into blank group, negative control group, RNAi S100A9group, and RNAi CyPA group after Interference,then calculating the clone forming rate and survival fraction by colony forming experiment after irradiation,Evaluating cell radiation sensitivity of RNAi silencing the S100A9, CyPA in lung cancer by Click multiple target model, and cell cycle analyzed by flow cytometry.
Result:Fuzheng Zengxiao formula has antitumor effects,which compared with the control group, Chinese medicine group,and radiation group,Tumor inhibition rate of traditional Chinese medicine+radiation group was striking increased.Results of Western blot showed:at6h after radiotherapy, that compared with control group, protein expression of S100A9and CyPA were decreased in each group, but no statistical difference.at12h after radiotherapy, Compared with the control group,expression of Chinese medicine+radiation group were decreased, and there were statistical significant difference(P<0.05).at24h after radiotherapy,The protein expression of S100A9,compared with the control group,radiation group was decreased,and there were statistical significant difference(P<0.05).To the other groups,expression of Chinese medicine+radiation group are decreased,there are significant.Co mpared with the control group,the protein expression of CyPA in the Chinese medicine group and radiation group was decreased,and there were statistical significant differences(P<0.05).T o the other groups,The protein expression of CyPA in Chinese medicine+radiation group were decreased,and there were statistical significant difference.Results of Real-time quantitative PCR show:at6h after radiotherapy,compared with control group,mRNA expression of S100A9and CyPA were decreased in the other group, but there were no difference. At12h after radiotherapy,Compared with the control group,mRNA expression of S100A9of radiation group were decreased,and there were statistical significant difference(P<0.05).Compared with the control group and Chinese medicine,Chinese medicine+radiation group were decreased,th ere are statistical significant difference (P<0.05).mRNA expression of CyPA,Compared with the control group,radiation group and Chinese medicine+radiation group are decreased,there are statistical significant difference(P<0.05).at24h after radiotherapy, mRNA expression of S100A9and CyPA, Compared with the control group、Chinese medicine,radiation group are decreased,there are statistical significant difference(P<0.05).To the other groups,expression of Chinese medicine+radiation group were decreased,there were statistical significant difference (P<0.05).Furthe, Construction of the lentiviral expression plasmid S100A9and CyPA RNAi gene and packing lentiviral.Select effective interference target sequence of S100A9and CyPA RNAi by Western blot,and stably transfected cells of S100A9and CyPA shRNA/PAa were prepared.It calculated the clone forming rate and survival fraction by the colony forming experiment,Using click multiple target model,it find that reduce SF2Gy value, Do value, Dq value of tumor cell after RNAi (S100A9,cyclophilin A),Increasing the radiation effect and enhance cytotoxic effects of radiation on tumour cells.The results of flow cytometry,after RNAi silencing S100A9and CyPA,without radiotherapy,Go/G1,S phase incells of every Groups had difference,but there were no statistical difference.G2/M phase in cells, Compared with the blank group, the S100A9group increased,there were statistical significant difference(P<0.05).Compared with the blank group and control group,CyPA group were increased,there were significant difference(P<0.05).After2Gy radiotherapy,Compared with the blank group, control group, G0/G1cells of S100A9and CyPA group were decreased,G2/M cells are statistical significanly increased(P<0.05).S cells of S100A9group were decreased,but no statistical difference.S cells of CyPA group were significantly decreased(P<0.05).
Conclusion:The experiment demonstrated that Fuzheng Zengxiao formula had certein damps the lump rate and better radiosensitizing effect on lung adenocarcinoma.Detection the expression of S100A9and CyPA by Western blot and q-PCR,it discoved that Fuzheng Zengxiao formula played radiosensitizing effect by down-regulating expression of S100A9and CyPA in lung adenocarcinoma.24h after radiotherapy,the express was significantly decreased,and there were statistical significant difference(P<0.05).Further,Construction of the lentiviral expression plasmid S100A9and CyPA RNAi gene,select effective interference target sequence by Western blot, and stably transfected cells of S100A9and CyPA shRNA/PAa were prepared.In vitro experiment,S100A9and CyPA siRNA mediated Lentivirus can improve ratio of G2/M phase in cell,and increase radiation sensitivity of cell.
引文
1.Jens Overgaard,Hanne Sand Hansen,Lena Specht,et al.Five compared with six fractions per week of conventional radiotherapy of squamous—cell carcinoma of Head and neck:DAHA NCA 6&7 randomised controlled trial[J].Lancet,2003,362(9388):933-940.
2.沈瑜,糜福顺.肿瘤放射生物学[M].北京：中国医药科技出版社,2002.254-258.
3.黄金昶,张代钊.恶性肿瘤放射增敏中药的研究[J].癌症进展杂志,2004,2(6)：446.
4.高喜源,李俊英.中药对抗放疗毒副作用的临床观察[J].中华实用中西医杂志,2004,4(17)：902.
5.胡定政.扶正抗癌方对60例晚期非小细胞肺癌放、化疗的增效解毒临床观察.陕西中医,2005,36(3)：209-211.
6.张培,胡丕丽.中西医结合治疗对肿瘤患者生存治疗的影响.中华肿瘤杂志,2003,25(3)：302.
7.王慧杰,王朝霞.扶正增效消噎汤并后程加速超分割放射治疗食管癌疗效观察.辽宁中医杂志,2004,31(5)：373-378.
8.黄金昶,张代钊.扶正增效方对肺腺癌细胞放射增敏研究[J].中华放射肿瘤学杂志,2000,9(2)：135-136.
9.黄金昶,张代钊,李佩文,等.扶正增效方配合肺癌放疗远期疗效分析[J].中国医刊,2000,35(1)：48.
10.黄静,李佩文,贾立群.扶正增效方配合放疗治疗非小细胞肺癌疗效观察[J].中日友好医院学报,2005,19(5)：278-280.
11.杨树明,张爱萍,蔡焦生.参芪六味地黄汤加放疗治疗食管癌的临床研究[J].光明中医,2008,23(2)：145-146.
12.刘爱荣.加减八珍汤对宫颈癌放疗增敏作用临床观察体会[J].卫生职业教育,2005,23(14)：127-128.
13.潘毅生,袁瑞香,黄克俊,等.仙鹤草抗肿瘤的实验研究.药学通报,1983,18：50-54.
14.史纪兰.鹤草酚对人肺腺癌细胞株辐射增敏作用的实验室研究.山东省医学科学院学报,1987,6：663-668.
15.韩俊庆,王兴文,等.鹤草酚对肺腺癌放射增敏的临床前瞻性研究.中华放射医学与防护杂志,2003,23(5)：355-357.
16.侯华新,黎丹戎,等.银杏叶多糖在肿瘤放射、化学治疗中的增敏作用研究[J].广西医科大学学报,2005,22(1)：29-31.
17.李毓.薏苡仁配合放射治疗晚期鼻咽癌的初步观察[J].桂林医学院学报,1997,10(1)：24-25.
18.李毓.薏苡仁配合放射治疗晚期鼻咽癌的远期疗效分析[J].华夏医学,1998,11(1)：10-11.
19.陈礼华,徐辉,等.薏苡仁提取物注射液联合放疗治疗恶性肿瘤的临床观察[J].肿瘤防治杂志,2000,7(6)：669.
20.李毓,胡笑克,熊带水,等.薏苡仁酯对人鼻咽癌细胞乏氧照射的增敏作用[J].中药新药与临床药理,2000,11(5)：269-271.
21.王琨,董惠芳,章晓鹰,等.夏枯草对SGC-7901细胞的影响[J].上海检验杂志,2000,15(5)：305-307.
22.张克杰,张明智,王庆端.夏枯草注射液诱导K562细胞凋亡的实验研究[J].中草药,2005,36(7)：1031-1035.
23.Horikawa K,Mohri T,Tanaka Y,et al.Moderate inhibition of mutagenicity and carcinogeni City of benzoapyrene,1,6-dinitropyrene and 3,9-dinitrofluoran theneby Chinese medicina 1 herbs.Mutagenesis,1994,9(6):523-528.
24.李甸源,邓晓明,高松涛,等.中西医结合治疗非小细胞肺癌脑转移33例[J].中医研究,2009,22(7)：37-38.
25.徐伯平,胡丕丽,黄国贤,等.中药散结方对鼻咽癌放疗增敏的研究[J].中医研究,2001,14(3)：17-19.
26.Wouter A,Pauwel B,Lardon F,at al.Implications of in vitro research on the effect of radiotherapy and chemotherapy under hypoxic conditions[J].Oncologist,2007,12(6):390-7 12.
27.Rockwell S,Dobrucki IT,Kim EY,et al.Hypoxia and raditiontherapy:past history; ongoing resea rch;and future promise[J].Curr Mol Med,2009,9(4):442-458.
28.Karar J,Maity A.Modulating the tumor microenvironment to increase radiationg respon siveness[J].Cancer Biol Ther,2009,(21):1994-2001.
29.史建军,邬晓东,刘锦全,等.中药增敏散配合放疗治疗鼻咽癌的疗效观察[J].国际医药卫生导报,2004,10(24)：4-5.
30.李伟雄,陈应瑞,徐国镇.活血化瘀中药联合放疗治疗鼻咽癌[J].广东医药,2002,23(1)：95-96.
31.黄光武,谢成熹,等.水蛭等活血化瘀中药与放疗联合治疗晚期鼻咽癌41例[J].中国中西医结合杂志,2003,23(10)：777-778.
32.张勇,白杰,王清朝.回生口服液加放疗治疗食管癌的远期疗效观察[J].河北医药,2000,22(5)：371.
33.万志龙,浦普言,马煌如.食管癌放射治疗并用中药活血化瘀的临床观察[J].南通医学院学报,1995,15(1)128-129.
34.曹璋,崔敏,孙宁.姜黄素对人鼻咽癌CEN-2Z细胞的放射增敏作用及其作用机制[J].现代肿瘤医学,2007,15(9)：1232-1233.
35.张晓智,韩苏夏,杨玉琮,等.姜黄素对食管癌细胞系放射敏感性的影响及其机理[J].西安交通大学学报(医学版),2003,24(4)：377-378.
36.王振华,师国珍,张宁,等.去甲斑蝥素对肺癌A549细胞株体外放射增敏作用[J].中国实用医刊,2009,36(24)：8。9.
37.Posmidis P,Mylonakis N.Paclitaxel and carboplatin in inopereable nonsmall cell lung.Semin Oncol,1996,23(16):7-10.
38.Machtay M,Aviles V,Kligerman MM,et al.A phase I trial of 96 hour paclitaxel infusion plus accelerated radiotherapy for unresectable head and neck cancer.IntJRadiat Oncol Biol Phys,1999,44(2):311315.
39.Hao Jiang,Shenglin Ma,Jianguo Feng.In vitro study of radiosensitization by P-Elemene in A549 cell line from adenocarcinoma of lung.Chinese-German Journal of Clinical Oncology,2009,8(1):12-15.
40.杨道科,宋一帆.β-榄香烯对人肝癌细胞株的体外放射增敏作用[J].中国现代药物应用,2008,2(17)：47-48.
41.吴大鹏,李新明,赵军方,等.β-榄香烯乳对舌鳞癌细胞株Tca-8113体外放射增敏观察[J].肿瘤基础与临床,2006,19(2)：116-117.
42.佘军军,王子明,车向明,等.p-榄香烯对兔VX2肾移植癌的放射增敏作用[J].中西医结合学报,2006,7(4)：392-396.
43.巴艳,庄兆虹,王燕.养阴解毒汤配合放射治疗鼻咽癌的临床观察[J].中医药学报,2010,38(4)：71-73.
44.王大海,田道法.益气解毒颗粒配合放射治疗鼻咽癌的临床研究[J].湖南中医学院学报,2006,26(1)：36-37.
45.冯俊光,乔冠英,林清,等.增液解毒颗粒干预放射性肺损伤[J].中国实验方剂学杂志,2011,17(10)：257-259.
46.司维柯,肖桃元,康格非.苦参碱对人肝癌细胞HepG2的细胞形态影响和相关增值因素的变化[J].第三军医大学学报,2000,272(6)：553-556.
47.杨光伟,陈洁,覃强,等.复方苦参合并放疗治疗鼻咽癌的临床观察[J].肿瘤研究与临床,2002,14(2)：124-126.
48.郑剑霄,周同冲,徐凯,等.石上柏联合放疗治疗鼻咽癌临床观察[J].南方医科大学学报,2006,26(2)：247-248.
49.李学源,王颖,于鲁华,等.马蔺子素放射增敏治疗食管癌的疗效观察[J].临床肿瘤学杂志,2000,5(4)：272-274.
50.吴庭安.马蔺子素配合放疗对鼻咽癌颈部淋巴结转移灶的近期疗效观察[J].基层医学论坛,2005,9(8)：673-674.
51.周益琴,卞秀华.马蔺子素在鼻咽癌放射中增敏作用的临床研究[J].南京医科大学学 报,2001,21(4)：328-330.
52.周厚清,刑成,李伯南.肿瘤放射增敏剂及增敏机制研究进展[J].国外医学肿瘤学分册,2001,28(6)：432-435.
53.李勤,谢艳华,孙纪元,等.中药安迪粉针剂的放射增敏作用[J].华西药学杂志,2003,18(1).
54.吴大鹏,姜睿斌,赵德强,等.β-榄香烯对体外培养Tca-8113细胞放射增敏、细胞周期、凋亡及Bcl-2、Bax蛋白表达的影响[J].郑州大学学报(医学版),2009,44(2)：414-417.
55.李明润,关俊玲,柴淑兰,等.Iq7611对离体培养的人宫颈癌细胞谷胱甘肽含量的影响—放射增敏机理探讨[J].中国肿瘤临床,1987,14(2)：92-94.
56.孙新臣,邓文英,李苏宜,等.粉防已碱对人肝癌细胞株7402的放射增敏作用[J].临床肿瘤学杂志,2007,12(12)：893-896.
57.杨道科,张转建,朱洪海.氧化苦参碱对宫颈癌细胞HeLa的放射增敏作用机制研究[J].中医研究,2008,21(8)：15-17.
1.Guo s, Kemphues KJ. Par-I, a gene for establishing polarity in C.elegans embryos, encodes a putative Ser/Thr kinase that is asymmetrically distributed[J]. Cell,1995,81(4):611-620.
2. Fire A, Xu S, Montgomery M K, et al. Potent and specific genetic interfer-ence by double-stranded RNA in Caenorhabditis elegans[J].Natrue,1998,391:806-811.
3.Leung RK, Whittaker PA. RNA interference:from gene silencing to gene-specific therapeuti cs[J]. Pharmacol Ther,2005,107:222-239.
4.Fu GF, Lin XH, Han QW, et al. RNA interference remarkably suppresses bcl-2 gene express ion in cancer cells in vitro and in vivo[J].Cancer Biol Ther,2005,4(8):822-829.
5.Miao GY, Lu QM, Zhang XL. Do w nregu 1 ation of surv i vi n by RNAi inhibits growth of human gastric carcinoma cells [J]. World J Gastroenterol,2007,13(8):1170-1174.
6.谯敏,向延秀,王丕龙.RNA干扰抑制裸鼠皮下移植瘤中Livin基因表达的实验研究.重庆医科大学学报,2010,35(2)：196-198.
7.Vanderlaag KE, HUDAK S, BALD L. Anterior gradient-2 plays a critical role in breast can cer cell growth and survival by modulating cyclin D1,estrogen receptor-alpha and survivin g [J]. Breast Cancer Res,2010,12(3):32-37.
8.KONG R, SUN B, WANG S J, et al. An experiment study of gemcitabine inducing pancreat ic cancer cell apoptosis potentiated by nuclear factor-kappa B P65 siRNA [J]. Chin J Surg(中华外科杂志),2010,48(2)：128-133.
9.ZHOU J J, CHEN M L, ZHANG Q Z. Blocking gastrin and CCK-B autocrine loop affects cell proliferation and apoptosis in vitro [J].Mol Cell Biochem,2010,343(1/2):133-141.
10.Verma U N, Surabhi R M, Schmaltieg A, et al. Small interfering RNAs directed against β-catenin inhibit the in vitro and in vivo growth of colon cancer cells[J]. Clin Cancer Res,2 003,9(4):1291-1300.
11.lei T, Han S, Guo XY, et al.The potential role of Id1 in cox-2 mediated angiogenesis in gas trie cancer [J].Zhong hua yi xue za zhi,2007,87(22):1570-1575.
12. Yin Y, Cao LY, Wu WQ, et al. Blocking effects of siRNA on VEGF expression in human colorectal cancer cells[J]. World J Gastroenterol,2010,16(9):1086-1092.
13.Guan H, Zhou Z, Wang H, et al. A small interfering RNA targeting vascular endothelial growth factor inhibits Ewing's sarcoma growth in a xenograft mouse model[J]. Clin Cancer Res,2005,11(7):2662-2669.
14.Kim WJ, Chang CW, Lee M, et al. Efficient siRNA delivery using water soluble lipopoly mer for antiangiogenie gene therapy[J].J Control Release,2007,118(3):357-363.
15.Wang W, Wang CY, Dong JH, et al. Identification of effective siRNA against K-ras in hu man pancreatic cancer cell line Mia.PaCa-2 by siRNA expression cassette[J]. World J Gast roenterol,2005,11 (13):2026-2031.
16.Jiang M, Milner J. Bcl-2 const itut ively suppresses P53 dependent apoptosis in colorectal cancer cells[J].Genes Dev,2003,17(7):832-837.
17.Liu O, Fu H, Xing R, et al. Survivin knockdown combined with apoptin over expression inhibits cell growth significantly [J]. Cancer Biol Ther,2008,7(7):1053-1060.
18.Liu Y, Chen L, Ko TC, et al. Evil is a survival factor which conveys resistance to both TGF beta and taxol-mediated cell death via PI3K/AKT[J]. Oncogene,2006,25(25):3565-3575.
19.Pichler A, Zelcer N, Prior JL, et al. In vivo RNA interference mediated ablation of MDR1 P-glycoprotein[J]. Clin Cancer Res,2005,11(12):4487-4494.
20.Leonard GD, Fojo T, Bates SE. The role of ABC transporters in clinical practice. Oncologi st,2003,8:411-424.
21.Yague E, Higgins CF, Raguz S. Complete reversal of multidrug resistance by stable expre ssion of small interfering RNAs targeting MDR1 [J].Gene Ther,2004,11:1170-1174.
22.Lou JY, Peng ZL, Zheng Y, et al. Research on human ovarian cancer cell MDR1 gene sile need by siRNA[J]. Sichuan Daxue Xuebao Yixueban,2007,38:753-755.
23.Lilleby W, Fossa SD. Chemotherapy in metastatic renal cell cancer[J]. World J Urol,2005, 23:175-179.
24.郝怡鑫,杜楠,肖文华.RNA干扰逆转肾癌细胞A498对长春新碱化疗耐药的研究[J].肿瘤研究与临床,2010,22(6)：391-395.
25.Patutina O, Mironova N, Popova N, et al. The siRNA targeted to mdr1b and mdr1a mRNA s in vivo sensitizes murine lymphosarcoma to chemotherapy [J]. BMC Cancer,2010,10(1) :204-207.
26.Susa M, Iyer AK, Ryu K, et al. Inhibition of ABCB1 (MDR1) expression by an siRNA nan Oparticulate delivery system to overcome drug resistance in osteosarcoma [J].PLoS One,2 010,5(5):764-769.
27.Wu H,Hait WN,rang JM.Small interfering RNA-induced suppression of MDR1(P-Glycopm tein)restores sensitivity to muhidrug resistant cancer cells [J].J Cancer Res,2003,63(7):15 15-1519.
28.沈自尹.基因科学和21世纪中医药学的走向[J].上海中医药杂志,2000,2(10)：7-8.
29.王颖芳,陈蔚文,劳绍贤,等.慢性胃炎脾气虚证与脾胃湿热证的差异表达基因比较[J].中国病理生理杂志,2008,24(2)：320-324.
30.王静.脾气虚证相关基因RPS20在IEC-6细胞的生物功能鉴定[D].广州：广州中医药大学,2009.
31.胡卫,方肇勤,梁超,等.RNAi研究角蛋白18在肝癌增殖中的作用及不同中医治法的调 节[J].安徽医药,2010,14(6)：632-634.
32.刘德麟,金香兰,杨威.从分子作用把握整体效应-中药分子药理的理论研究[J].自然杂志,1997,19(6)：316-323.
1.殷蔚伯,谷铣之.肿瘤放射治疗学[M].北京：中国协和医科大学出版社,2002：677.
2.吴大鹏,姜睿斌,赵德强,等.p-榄香烯对体外培养Tca-8113细胞放射增敏、细胞周期、凋亡及Bcl-2、Bax蛋白表达的影响[J].郑州大学学报(医学版),2009,44(2)：414-417.
3.周厚清,刑成,李伯南.肿瘤放射增敏剂及增敏机制研究进展[J].国外医学肿瘤学册,2001,28(6)：432-435.
4.Yui S,Nakatami Y,Mikami M.Calprotectin(S100A8/S100A9).an inflammatory protein com plex from neutrophils with a broad apoptosis inducing activity[J].Biol Pha rm Bull,2003,26: 753-760.
5.Arai K,Teratani T,Nozawa R,et al.Immunohistochemical investigation of S100A9 expressio n in pulmonary adenocarcinoma:S100A9 expression is associated with tumor differentiatio n. OncolRep,2001,8(3):591-596.
6.Visser KE,Eichten A,Coussens LM.Paradoxical roles of the immune system during cancer development[J].Nat Rev Cancer,2006,6(1):24-37.
7.Coussens LM,Werb Z,Inflammation and cancer[J].Nature,2002,420(6917):860-867.
8.Mueller MM,Fusening NE.Friends or foes-bipolar effects of the tumour stroma in cancer[J] .Nat RevCancer,2004,4(11):839-849.
9.Wang J,Cai Y,Xu H,et al. Expression of MRP14 gene is frequently down regulated inChine se human esophageal cancer[J].Cell Res,2004,14:46-53.
10.Roesch EM,Nees M,Karsai S,et al. Transcript and proteome analysis reveals reduced exp ression of calgranulins in head and neck squamous cell carcinoma[J]. Eur J Cell Biol,200 5,8 (2-3):431-441.
11.Camps MJ,Wang MZ,Howard B,et al.Protein expression profiling identifies macrophagem igration inhibitory factor and cyclophilin a as potential molecular targets in nonsmall cell lung cancer[J]. Cancer Res.2003,63 (7):1652-1656.
12.Howard BA,Furumai R,Campa MJ,et a 1.Stable RNA interference-mediated suppression of cyclophilin a diminishes non-small-cell lung tumor growth in vivo[J]. Cancer Res.2005 ,65 (19):8853-8860.
13.Shen J, Person MD, Zhu J, et al. Protein expression profiles in pancreatic adenocarcinoms compared with normal pancreatic tissue and tissue affected by pancreatitis as detected by tw o-dimensional gel electrophoresis and mass spectrometry[J].Cancer Res.2004,64 (24):90 18-9026.
14.Lim SO,Park SJ,Kim W,et al.Proteome analysis of hepatocellular carcinoma[J]. Biochem Biophys Res Commun.2002,291 (4):1031-1037.
15.Zheng J,Koblinski JE,Dutson L V,et al.Prolyl isomerase cyclophilin a regulation of Janus-activated kinase 2 and the progression of human breast cancer[J]. Cancer Res.2008,68 (19):7769-7778.
16. AI-Ghoul,Bruck TB.Lauer-Fields JL,et al. Comparative proteomic analysis of matched primary and metastatic melanoma cell lines[J]. J Proteome Res,2008,7(9):4107-4118.
1.Lagasse E, Weissman IL. Mouse MRP8 and MRP 14, two intracellular calcium-binding pro teins associated with the development of the myeloid lineage.Blood 1992;79(8):1907-15.
2.El-Rifai W, Moskaluk CA, Abdrabbo MK, Harper J, Yoshida C, Riggins GJ.et al. Gastric cancers overexpress S100A calcium-binding proteins. Cancer Res 2002; 62(23):6823-6.
3.Kawai H, Minamiya Y, Takahashi N. Prognostic impact of S100A9 overexpression in no n-small cell lung cancer. Tumour Biol;32(4):641-6.
4.Kim HJ, Kang HJ, Lee H, Lee ST, Yu MH, Kim H, et al. Identification of S100A8 and S100A9 as serological markers for colorectal cancer.J Proteome Res 2009;8(3):1368-79.
5.Acharyya S, Oskarsson T, Vanharanta S, Malladi S, Kim J, Morris PG, et al.A CXCL1 paracr inenetwork links cancer chemoresistance and metastasis.Cell;150(1):165-78.
6.Dorfman T, Weimann A, Borsetti A, Walsh CT, Gottlinger HG. Active-site residues of cy clophil in A are crucial for its incorporation into human immunodeficiency virus type 1 vi rions. J Virol 1997;71(9):7110-3.
7.Bosco DA, Eisenmesser EZ, Pochapsky S, Sundquist WI, Kern D. Catalysis of cis/transi some rization in native HIV-1 capsid by human cyclophilin A. Proc Natl Acad Sci USA 2002;99(8):5247-52.
8.Fire A, Xu S, Montgomery MK, Kostas SA, Driver SE, Mello CC. Potent and specific gen etic interference by double-stranded RNA in Caenorhabditis elegans.Nature1998;391(666 9):806-11.
9.Castel SE, Martienssen RA. RNA interference in the nucleus:roles for small RNAs in rans cri tption, epigenetics and beyond. Nat Rev Genet; 14(2):100-12.
10.Qi Y, Denli AM, Hannon GJ. Biochemical specialization within Arabidopsis RNA silenci ng pathways. Mol Cell 2005;19(3):421-8.
11.Mi S, Cai T, Hu Y, Chen Y, Hodges E, Ni F, et al. Sorting of small RNAs into Arabidopsis argonaute complexes is directed by the 5'terminal nucleotide. Cell 2008; 133(1):116-27.
12.Sledz CA, Williams BR.RNA interference in biology and disease.Blood 2005; 106 (3):787-94.
13.Rudnicki A, Avraham KB.microRNAs:the art of silencing in the ear. EMBO Mol Med;4 (9):849-59.
14.Eicher SC, Dehio C.Systems-level analysis of host-pathogen interaction using RNA interf erence.N Biotechnol.
15.Kim DH, Rossi JJ.Strategies for silencing human disease using RNA interference. Nat Rev Genet 2007;8(3):173-84.
16.Kato M,Slack FJ.Ageing and the small,non-coding RNA world.Ageing Res Rev; 12(1):42 9-35.
17.Rubinson DA,Dillon CP,Kwiatkowski AV,Sievers C,Yang L,Kopinja J,et al.A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and tra ns genic mice by RNA interference.Nat Genet 2003;33 (3):401-6.
1.H iratsuka S, W atanabe A, Aburatan iH, et al.Tumour-mediated upregulation of chemoat tr act ants and recruitment of myeloid cells predetermines lung metas tasis[J].Nat Cell Bio,120 06,8(12):1369-1375.
2.Choi KJ,Piao YJ,Lim MJ,et al.Overexpressed cyclophilin A in cancer cells renders esist anc e to hypoxia-and cisplatin-induced cell death.Cancer Res,2007.67:3654-3662.
3.Mul Ian PB. Quinn JE. Harkin DP. The role of BRCA1 in transcriptional regulation and xel 1 cycle control[J]. Oncogene,2006,25(43):5854-5863.
4.Nair S,Ram RR,Srinivas P, et al.Radiosensitizing effects of plumbagin in cervic al cancer ce lls is through modulation of apoptotic pathway. Mol Carcinoq,2008,47(1):22-33.
2.沈瑜,糜福顺.肿瘤放射生物学[M].北京：中国医药科技出版社,2002.254-258.
3.黄金昶,张代钊.恶性肿瘤放射增敏中药的研究[J].癌症进展杂志,2004,2(6)：446.
4.高喜源,李俊英.中药对抗放疗毒副作用的临床观察[J].中华实用中西医杂志,2004,4(17)：902.
5.胡定政.扶正抗癌方对60例晚期非小细胞肺癌放、化疗的增效解毒临床观察.陕西中医,2005,36(3)：209-211.
6.张培,胡丕丽.中西医结合治疗对肿瘤患者生存治疗的影响.中华肿瘤杂志,2003,25(3)：302.
7.王慧杰,王朝霞.扶正增效消噎汤并后程加速超分割放射治疗食管癌疗效观察.辽宁中医杂志,2004,31(5)：373-378.
8.黄金昶,张代钊.扶正增效方对肺腺癌细胞放射增敏研究[J].中华放射肿瘤学杂志,2000,9(2)：135-136.
9.黄金昶,张代钊,李佩文,等.扶正增效方配合肺癌放疗远期疗效分析[J].中国医刊,2000,35(1)：48.
10.黄静,李佩文,贾立群.扶正增效方配合放疗治疗非小细胞肺癌疗效观察[J].中日友好医院学报,2005,19(5)：278-280.
11.杨树明,张爱萍,蔡焦生.参芪六味地黄汤加放疗治疗食管癌的临床研究[J].光明中医,2008,23(2)：145-146.
12.刘爱荣.加减八珍汤对宫颈癌放疗增敏作用临床观察体会[J].卫生职业教育,2005,23(14)：127-128.
13.潘毅生,袁瑞香,黄克俊,等.仙鹤草抗肿瘤的实验研究.药学通报,1983,18：50-54.
14.史纪兰.鹤草酚对人肺腺癌细胞株辐射增敏作用的实验室研究.山东省医学科学院学报,1987,6：663-668.
15.韩俊庆,王兴文,等.鹤草酚对肺腺癌放射增敏的临床前瞻性研究.中华放射医学与防护杂志,2003,23(5)：355-357.
16.侯华新,黎丹戎,等.银杏叶多糖在肿瘤放射、化学治疗中的增敏作用研究[J].广西医科大学学报,2005,22(1)：29-31.
17.李毓.薏苡仁配合放射治疗晚期鼻咽癌的初步观察[J].桂林医学院学报,1997,10(1)：24-25.
18.李毓.薏苡仁配合放射治疗晚期鼻咽癌的远期疗效分析[J].华夏医学,1998,11(1)：10-11.
19.陈礼华,徐辉,等.薏苡仁提取物注射液联合放疗治疗恶性肿瘤的临床观察[J].肿瘤防治杂志,2000,7(6)：669.
20.李毓,胡笑克,熊带水,等.薏苡仁酯对人鼻咽癌细胞乏氧照射的增敏作用[J].中药新药与临床药理,2000,11(5)：269-271.
21.王琨,董惠芳,章晓鹰,等.夏枯草对SGC-7901细胞的影响[J].上海检验杂志,2000,15(5)：305-307.
22.张克杰,张明智,王庆端.夏枯草注射液诱导K562细胞凋亡的实验研究[J].中草药,2005,36(7)：1031-1035.
23.Horikawa K,Mohri T,Tanaka Y,et al.Moderate inhibition of mutagenicity and carcinogeni City of benzoapyrene,1,6-dinitropyrene and 3,9-dinitrofluoran theneby Chinese medicina 1 herbs.Mutagenesis,1994,9(6):523-528.
24.李甸源,邓晓明,高松涛,等.中西医结合治疗非小细胞肺癌脑转移33例[J].中医研究,2009,22(7)：37-38.
25.徐伯平,胡丕丽,黄国贤,等.中药散结方对鼻咽癌放疗增敏的研究[J].中医研究,2001,14(3)：17-19.
26.Wouter A,Pauwel B,Lardon F,at al.Implications of in vitro research on the effect of radiotherapy and chemotherapy under hypoxic conditions[J].Oncologist,2007,12(6):390-7 12.
27.Rockwell S,Dobrucki IT,Kim EY,et al.Hypoxia and raditiontherapy:past history; ongoing resea rch;and future promise[J].Curr Mol Med,2009,9(4):442-458.
28.Karar J,Maity A.Modulating the tumor microenvironment to increase radiationg respon siveness[J].Cancer Biol Ther,2009,(21):1994-2001.
29.史建军,邬晓东,刘锦全,等.中药增敏散配合放疗治疗鼻咽癌的疗效观察[J].国际医药卫生导报,2004,10(24)：4-5.
30.李伟雄,陈应瑞,徐国镇.活血化瘀中药联合放疗治疗鼻咽癌[J].广东医药,2002,23(1)：95-96.
31.黄光武,谢成熹,等.水蛭等活血化瘀中药与放疗联合治疗晚期鼻咽癌41例[J].中国中西医结合杂志,2003,23(10)：777-778.
32.张勇,白杰,王清朝.回生口服液加放疗治疗食管癌的远期疗效观察[J].河北医药,2000,22(5)：371.
33.万志龙,浦普言,马煌如.食管癌放射治疗并用中药活血化瘀的临床观察[J].南通医学院学报,1995,15(1)128-129.
34.曹璋,崔敏,孙宁.姜黄素对人鼻咽癌CEN-2Z细胞的放射增敏作用及其作用机制[J].现代肿瘤医学,2007,15(9)：1232-1233.
35.张晓智,韩苏夏,杨玉琮,等.姜黄素对食管癌细胞系放射敏感性的影响及其机理[J].西安交通大学学报(医学版),2003,24(4)：377-378.
36.王振华,师国珍,张宁,等.去甲斑蝥素对肺癌A549细胞株体外放射增敏作用[J].中国实用医刊,2009,36(24)：8。9.
37.Posmidis P,Mylonakis N.Paclitaxel and carboplatin in inopereable nonsmall cell lung.Semin Oncol,1996,23(16):7-10.
38.Machtay M,Aviles V,Kligerman MM,et al.A phase I trial of 96 hour paclitaxel infusion plus accelerated radiotherapy for unresectable head and neck cancer.IntJRadiat Oncol Biol Phys,1999,44(2):311315.
39.Hao Jiang,Shenglin Ma,Jianguo Feng.In vitro study of radiosensitization by P-Elemene in A549 cell line from adenocarcinoma of lung.Chinese-German Journal of Clinical Oncology,2009,8(1):12-15.
40.杨道科,宋一帆.β-榄香烯对人肝癌细胞株的体外放射增敏作用[J].中国现代药物应用,2008,2(17)：47-48.
41.吴大鹏,李新明,赵军方,等.β-榄香烯乳对舌鳞癌细胞株Tca-8113体外放射增敏观察[J].肿瘤基础与临床,2006,19(2)：116-117.
42.佘军军,王子明,车向明,等.p-榄香烯对兔VX2肾移植癌的放射增敏作用[J].中西医结合学报,2006,7(4)：392-396.
43.巴艳,庄兆虹,王燕.养阴解毒汤配合放射治疗鼻咽癌的临床观察[J].中医药学报,2010,38(4)：71-73.
44.王大海,田道法.益气解毒颗粒配合放射治疗鼻咽癌的临床研究[J].湖南中医学院学报,2006,26(1)：36-37.
45.冯俊光,乔冠英,林清,等.增液解毒颗粒干预放射性肺损伤[J].中国实验方剂学杂志,2011,17(10)：257-259.
46.司维柯,肖桃元,康格非.苦参碱对人肝癌细胞HepG2的细胞形态影响和相关增值因素的变化[J].第三军医大学学报,2000,272(6)：553-556.
47.杨光伟,陈洁,覃强,等.复方苦参合并放疗治疗鼻咽癌的临床观察[J].肿瘤研究与临床,2002,14(2)：124-126.
48.郑剑霄,周同冲,徐凯,等.石上柏联合放疗治疗鼻咽癌临床观察[J].南方医科大学学报,2006,26(2)：247-248.
49.李学源,王颖,于鲁华,等.马蔺子素放射增敏治疗食管癌的疗效观察[J].临床肿瘤学杂志,2000,5(4)：272-274.
50.吴庭安.马蔺子素配合放疗对鼻咽癌颈部淋巴结转移灶的近期疗效观察[J].基层医学论坛,2005,9(8)：673-674.
51.周益琴,卞秀华.马蔺子素在鼻咽癌放射中增敏作用的临床研究[J].南京医科大学学 报,2001,21(4)：328-330.
52.周厚清,刑成,李伯南.肿瘤放射增敏剂及增敏机制研究进展[J].国外医学肿瘤学分册,2001,28(6)：432-435.
53.李勤,谢艳华,孙纪元,等.中药安迪粉针剂的放射增敏作用[J].华西药学杂志,2003,18(1).
54.吴大鹏,姜睿斌,赵德强,等.β-榄香烯对体外培养Tca-8113细胞放射增敏、细胞周期、凋亡及Bcl-2、Bax蛋白表达的影响[J].郑州大学学报(医学版),2009,44(2)：414-417.
55.李明润,关俊玲,柴淑兰,等.Iq7611对离体培养的人宫颈癌细胞谷胱甘肽含量的影响—放射增敏机理探讨[J].中国肿瘤临床,1987,14(2)：92-94.
56.孙新臣,邓文英,李苏宜,等.粉防已碱对人肝癌细胞株7402的放射增敏作用[J].临床肿瘤学杂志,2007,12(12)：893-896.
57.杨道科,张转建,朱洪海.氧化苦参碱对宫颈癌细胞HeLa的放射增敏作用机制研究[J].中医研究,2008,21(8)：15-17.
1.Guo s, Kemphues KJ. Par-I, a gene for establishing polarity in C.elegans embryos, encodes a putative Ser/Thr kinase that is asymmetrically distributed[J]. Cell,1995,81(4):611-620.
2. Fire A, Xu S, Montgomery M K, et al. Potent and specific genetic interfer-ence by double-stranded RNA in Caenorhabditis elegans[J].Natrue,1998,391:806-811.
3.Leung RK, Whittaker PA. RNA interference:from gene silencing to gene-specific therapeuti cs[J]. Pharmacol Ther,2005,107:222-239.
4.Fu GF, Lin XH, Han QW, et al. RNA interference remarkably suppresses bcl-2 gene express ion in cancer cells in vitro and in vivo[J].Cancer Biol Ther,2005,4(8):822-829.
5.Miao GY, Lu QM, Zhang XL. Do w nregu 1 ation of surv i vi n by RNAi inhibits growth of human gastric carcinoma cells [J]. World J Gastroenterol,2007,13(8):1170-1174.
6.谯敏,向延秀,王丕龙.RNA干扰抑制裸鼠皮下移植瘤中Livin基因表达的实验研究.重庆医科大学学报,2010,35(2)：196-198.
7.Vanderlaag KE, HUDAK S, BALD L. Anterior gradient-2 plays a critical role in breast can cer cell growth and survival by modulating cyclin D1,estrogen receptor-alpha and survivin g [J]. Breast Cancer Res,2010,12(3):32-37.
8.KONG R, SUN B, WANG S J, et al. An experiment study of gemcitabine inducing pancreat ic cancer cell apoptosis potentiated by nuclear factor-kappa B P65 siRNA [J]. Chin J Surg(中华外科杂志),2010,48(2)：128-133.
9.ZHOU J J, CHEN M L, ZHANG Q Z. Blocking gastrin and CCK-B autocrine loop affects cell proliferation and apoptosis in vitro [J].Mol Cell Biochem,2010,343(1/2):133-141.
10.Verma U N, Surabhi R M, Schmaltieg A, et al. Small interfering RNAs directed against β-catenin inhibit the in vitro and in vivo growth of colon cancer cells[J]. Clin Cancer Res,2 003,9(4):1291-1300.
11.lei T, Han S, Guo XY, et al.The potential role of Id1 in cox-2 mediated angiogenesis in gas trie cancer [J].Zhong hua yi xue za zhi,2007,87(22):1570-1575.
12. Yin Y, Cao LY, Wu WQ, et al. Blocking effects of siRNA on VEGF expression in human colorectal cancer cells[J]. World J Gastroenterol,2010,16(9):1086-1092.
13.Guan H, Zhou Z, Wang H, et al. A small interfering RNA targeting vascular endothelial growth factor inhibits Ewing's sarcoma growth in a xenograft mouse model[J]. Clin Cancer Res,2005,11(7):2662-2669.
14.Kim WJ, Chang CW, Lee M, et al. Efficient siRNA delivery using water soluble lipopoly mer for antiangiogenie gene therapy[J].J Control Release,2007,118(3):357-363.
15.Wang W, Wang CY, Dong JH, et al. Identification of effective siRNA against K-ras in hu man pancreatic cancer cell line Mia.PaCa-2 by siRNA expression cassette[J]. World J Gast roenterol,2005,11 (13):2026-2031.
16.Jiang M, Milner J. Bcl-2 const itut ively suppresses P53 dependent apoptosis in colorectal cancer cells[J].Genes Dev,2003,17(7):832-837.
17.Liu O, Fu H, Xing R, et al. Survivin knockdown combined with apoptin over expression inhibits cell growth significantly [J]. Cancer Biol Ther,2008,7(7):1053-1060.
18.Liu Y, Chen L, Ko TC, et al. Evil is a survival factor which conveys resistance to both TGF beta and taxol-mediated cell death via PI3K/AKT[J]. Oncogene,2006,25(25):3565-3575.
19.Pichler A, Zelcer N, Prior JL, et al. In vivo RNA interference mediated ablation of MDR1 P-glycoprotein[J]. Clin Cancer Res,2005,11(12):4487-4494.
20.Leonard GD, Fojo T, Bates SE. The role of ABC transporters in clinical practice. Oncologi st,2003,8:411-424.
21.Yague E, Higgins CF, Raguz S. Complete reversal of multidrug resistance by stable expre ssion of small interfering RNAs targeting MDR1 [J].Gene Ther,2004,11:1170-1174.
22.Lou JY, Peng ZL, Zheng Y, et al. Research on human ovarian cancer cell MDR1 gene sile need by siRNA[J]. Sichuan Daxue Xuebao Yixueban,2007,38:753-755.
23.Lilleby W, Fossa SD. Chemotherapy in metastatic renal cell cancer[J]. World J Urol,2005, 23:175-179.
24.郝怡鑫,杜楠,肖文华.RNA干扰逆转肾癌细胞A498对长春新碱化疗耐药的研究[J].肿瘤研究与临床,2010,22(6)：391-395.
25.Patutina O, Mironova N, Popova N, et al. The siRNA targeted to mdr1b and mdr1a mRNA s in vivo sensitizes murine lymphosarcoma to chemotherapy [J]. BMC Cancer,2010,10(1) :204-207.
26.Susa M, Iyer AK, Ryu K, et al. Inhibition of ABCB1 (MDR1) expression by an siRNA nan Oparticulate delivery system to overcome drug resistance in osteosarcoma [J].PLoS One,2 010,5(5):764-769.
27.Wu H,Hait WN,rang JM.Small interfering RNA-induced suppression of MDR1(P-Glycopm tein)restores sensitivity to muhidrug resistant cancer cells [J].J Cancer Res,2003,63(7):15 15-1519.
28.沈自尹.基因科学和21世纪中医药学的走向[J].上海中医药杂志,2000,2(10)：7-8.
29.王颖芳,陈蔚文,劳绍贤,等.慢性胃炎脾气虚证与脾胃湿热证的差异表达基因比较[J].中国病理生理杂志,2008,24(2)：320-324.
30.王静.脾气虚证相关基因RPS20在IEC-6细胞的生物功能鉴定[D].广州：广州中医药大学,2009.
31.胡卫,方肇勤,梁超,等.RNAi研究角蛋白18在肝癌增殖中的作用及不同中医治法的调 节[J].安徽医药,2010,14(6)：632-634.
32.刘德麟,金香兰,杨威.从分子作用把握整体效应-中药分子药理的理论研究[J].自然杂志,1997,19(6)：316-323.
1.殷蔚伯,谷铣之.肿瘤放射治疗学[M].北京：中国协和医科大学出版社,2002：677.
2.吴大鹏,姜睿斌,赵德强,等.p-榄香烯对体外培养Tca-8113细胞放射增敏、细胞周期、凋亡及Bcl-2、Bax蛋白表达的影响[J].郑州大学学报(医学版),2009,44(2)：414-417.
3.周厚清,刑成,李伯南.肿瘤放射增敏剂及增敏机制研究进展[J].国外医学肿瘤学册,2001,28(6)：432-435.
4.Yui S,Nakatami Y,Mikami M.Calprotectin(S100A8/S100A9).an inflammatory protein com plex from neutrophils with a broad apoptosis inducing activity[J].Biol Pha rm Bull,2003,26: 753-760.
5.Arai K,Teratani T,Nozawa R,et al.Immunohistochemical investigation of S100A9 expressio n in pulmonary adenocarcinoma:S100A9 expression is associated with tumor differentiatio n. OncolRep,2001,8(3):591-596.
6.Visser KE,Eichten A,Coussens LM.Paradoxical roles of the immune system during cancer development[J].Nat Rev Cancer,2006,6(1):24-37.
7.Coussens LM,Werb Z,Inflammation and cancer[J].Nature,2002,420(6917):860-867.
8.Mueller MM,Fusening NE.Friends or foes-bipolar effects of the tumour stroma in cancer[J] .Nat RevCancer,2004,4(11):839-849.
9.Wang J,Cai Y,Xu H,et al. Expression of MRP14 gene is frequently down regulated inChine se human esophageal cancer[J].Cell Res,2004,14:46-53.
10.Roesch EM,Nees M,Karsai S,et al. Transcript and proteome analysis reveals reduced exp ression of calgranulins in head and neck squamous cell carcinoma[J]. Eur J Cell Biol,200 5,8 (2-3):431-441.
11.Camps MJ,Wang MZ,Howard B,et al.Protein expression profiling identifies macrophagem igration inhibitory factor and cyclophilin a as potential molecular targets in nonsmall cell lung cancer[J]. Cancer Res.2003,63 (7):1652-1656.
12.Howard BA,Furumai R,Campa MJ,et a 1.Stable RNA interference-mediated suppression of cyclophilin a diminishes non-small-cell lung tumor growth in vivo[J]. Cancer Res.2005 ,65 (19):8853-8860.
13.Shen J, Person MD, Zhu J, et al. Protein expression profiles in pancreatic adenocarcinoms compared with normal pancreatic tissue and tissue affected by pancreatitis as detected by tw o-dimensional gel electrophoresis and mass spectrometry[J].Cancer Res.2004,64 (24):90 18-9026.
14.Lim SO,Park SJ,Kim W,et al.Proteome analysis of hepatocellular carcinoma[J]. Biochem Biophys Res Commun.2002,291 (4):1031-1037.
15.Zheng J,Koblinski JE,Dutson L V,et al.Prolyl isomerase cyclophilin a regulation of Janus-activated kinase 2 and the progression of human breast cancer[J]. Cancer Res.2008,68 (19):7769-7778.
16. AI-Ghoul,Bruck TB.Lauer-Fields JL,et al. Comparative proteomic analysis of matched primary and metastatic melanoma cell lines[J]. J Proteome Res,2008,7(9):4107-4118.
1.Lagasse E, Weissman IL. Mouse MRP8 and MRP 14, two intracellular calcium-binding pro teins associated with the development of the myeloid lineage.Blood 1992;79(8):1907-15.
2.El-Rifai W, Moskaluk CA, Abdrabbo MK, Harper J, Yoshida C, Riggins GJ.et al. Gastric cancers overexpress S100A calcium-binding proteins. Cancer Res 2002; 62(23):6823-6.
3.Kawai H, Minamiya Y, Takahashi N. Prognostic impact of S100A9 overexpression in no n-small cell lung cancer. Tumour Biol;32(4):641-6.
4.Kim HJ, Kang HJ, Lee H, Lee ST, Yu MH, Kim H, et al. Identification of S100A8 and S100A9 as serological markers for colorectal cancer.J Proteome Res 2009;8(3):1368-79.
5.Acharyya S, Oskarsson T, Vanharanta S, Malladi S, Kim J, Morris PG, et al.A CXCL1 paracr inenetwork links cancer chemoresistance and metastasis.Cell;150(1):165-78.
6.Dorfman T, Weimann A, Borsetti A, Walsh CT, Gottlinger HG. Active-site residues of cy clophil in A are crucial for its incorporation into human immunodeficiency virus type 1 vi rions. J Virol 1997;71(9):7110-3.
7.Bosco DA, Eisenmesser EZ, Pochapsky S, Sundquist WI, Kern D. Catalysis of cis/transi some rization in native HIV-1 capsid by human cyclophilin A. Proc Natl Acad Sci USA 2002;99(8):5247-52.
8.Fire A, Xu S, Montgomery MK, Kostas SA, Driver SE, Mello CC. Potent and specific gen etic interference by double-stranded RNA in Caenorhabditis elegans.Nature1998;391(666 9):806-11.
9.Castel SE, Martienssen RA. RNA interference in the nucleus:roles for small RNAs in rans cri tption, epigenetics and beyond. Nat Rev Genet; 14(2):100-12.
10.Qi Y, Denli AM, Hannon GJ. Biochemical specialization within Arabidopsis RNA silenci ng pathways. Mol Cell 2005;19(3):421-8.
11.Mi S, Cai T, Hu Y, Chen Y, Hodges E, Ni F, et al. Sorting of small RNAs into Arabidopsis argonaute complexes is directed by the 5'terminal nucleotide. Cell 2008; 133(1):116-27.
12.Sledz CA, Williams BR.RNA interference in biology and disease.Blood 2005; 106 (3):787-94.
13.Rudnicki A, Avraham KB.microRNAs:the art of silencing in the ear. EMBO Mol Med;4 (9):849-59.
14.Eicher SC, Dehio C.Systems-level analysis of host-pathogen interaction using RNA interf erence.N Biotechnol.
15.Kim DH, Rossi JJ.Strategies for silencing human disease using RNA interference. Nat Rev Genet 2007;8(3):173-84.
16.Kato M,Slack FJ.Ageing and the small,non-coding RNA world.Ageing Res Rev; 12(1):42 9-35.
17.Rubinson DA,Dillon CP,Kwiatkowski AV,Sievers C,Yang L,Kopinja J,et al.A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and tra ns genic mice by RNA interference.Nat Genet 2003;33 (3):401-6.
1.H iratsuka S, W atanabe A, Aburatan iH, et al.Tumour-mediated upregulation of chemoat tr act ants and recruitment of myeloid cells predetermines lung metas tasis[J].Nat Cell Bio,120 06,8(12):1369-1375.
2.Choi KJ,Piao YJ,Lim MJ,et al.Overexpressed cyclophilin A in cancer cells renders esist anc e to hypoxia-and cisplatin-induced cell death.Cancer Res,2007.67:3654-3662.
3.Mul Ian PB. Quinn JE. Harkin DP. The role of BRCA1 in transcriptional regulation and xel 1 cycle control[J]. Oncogene,2006,25(43):5854-5863.
4.Nair S,Ram RR,Srinivas P, et al.Radiosensitizing effects of plumbagin in cervic al cancer ce lls is through modulation of apoptotic pathway. Mol Carcinoq,2008,47(1):22-33.