【作者】 李渊渊；

【导师】 钟玲；

【作者基本信息】 重庆医科大学 ， 妇产科学， 2010， 硕士

【摘要】 目的(1)观察顺铂导致人卵巢癌SKOV3细胞药物耐受后细胞凋亡情况。探讨DNA错配修复基因hMLH1、hMSH2的在人卵巢癌细胞SKOV3及耐顺铂人卵巢癌细胞SKOV3/DDP中的转录与翻译表达变化。(2)探讨耐顺铂人卵巢癌细胞SKOV3/DDP中凋亡基因bax、bcl-2、caspase9的转录与翻译表达变化,为进一步研究凋亡途径异常提供实验室依据。方法(1)在第一部分细胞培养实验中,采用MTT法、Hoechst免疫荧光染色观察顺铂对卵巢癌SKOV3细胞和卵巢癌顺铂耐药SKOV3/DDP细胞凋亡情况,检测SKOV3/DDP细胞耐药性。使用RT-PCR技术及Western blotting技术分别检测顺铂敏感性人卵巢癌SKOV3细胞及耐顺铂人卵巢癌SKOV3/DDP细胞中错配修复基因hMLH1和hMSH2的转录与翻译表达变化情况。(2)在第二部分实验中,使用RT-PCR技术及Western-blotting技术分别检测顺铂敏感性人卵巢癌SKOV3细胞及耐顺铂人卵巢癌SKOV3/DDP细胞中凋亡相关基因bcl-2、bax、caspase9的转录与翻译表达变化情况。结果(1)同一顺铂浓度作用下,耐顺铂人卵巢癌细胞的凋亡率明显低于顺铂敏感性人卵巢癌细胞。顺铂敏感性人卵巢癌细胞的错配修复基因hMLH1和hMSH2的mRNA及蛋白表达水平均显著高于耐顺铂人卵巢癌细胞。(2)顺铂敏感性人卵巢癌细胞的凋亡相关基因bax、caspase9的mRNA及蛋白表达水平均显著高于耐顺铂人卵巢癌细胞,而bcl-2的mRNA及蛋白表达水平反而在耐顺铂人卵巢癌细胞有明显增高。结论错配修复基因hMLH1和hMSH2在耐顺铂人卵巢癌细胞中低表达可能是导致顺铂耐药发生的重要原因。凋亡基因bcl-2、bax、caspase9在耐顺铂人卵巢癌细胞中异常表达可能与顺铂耐药的发生有直接相关性。更多还原

【Abstract】 Objective (1)Observation of cisplatin led to SKOV3 human ovarian cancer cell apoptosis after drug resistance。To investigate the roles of hMLH1 and hMSH2 gene in cisplatin resistance in human ovarian cancer. (2)To investigate the roles of bax,bcl-2 and caspase9 gene in cisplatin resistance in human ovarian cancer. To provide the apoptosis way for laboratory further study.Methods (1)The chemosensitivity to cisplatin was measured by MTT assay. The apoptosis induced by DDP was tested by Hoechst fluorescence analysis. The gene expression of hMLH1 and hMLH2 at transcription and translation levels in SKOV3 and SKOV3/DDP were measured by reverse transcription polymerase chain reaction (RT-PCR) and Western blotting, respectively. (2)The gene expression of bax,bcl-2 and caspase9 at transcription and translation levels in SKOV3 and SKOV3/DDP were measured by reverse transcription polymerase chain reaction (RT-PCR) and Western blotting, respectively.Results (1)The apopototic rate of SKOV3/DDP was markedly reduced compared to that of SKOV3 after cisplatin treatment. Both mRNA and protein levles of hMSH2 in SKOV3 were significantly higher than those observed in SKOV3/DDP. (2)Both mRNA and protein levles of bax and caspase9 in SKOV3 were significantly higher than those observed in SKOV3/DDP. mRNA and protein levles of bcl-2 in SKOV3 were significantly lower than those observed in SKOV3/DDPConclusion The inhibition of hMLH1 and hMSH2 gene expression may contribute to the occurrence of cisplatin resistance in human ovarian cancer.Also The inhibition of bax,bcl-2 and caspase9 gene expression may contribute to the occurrence of cisplatin resistance in human ovarian cancer更多还原