论文部分内容阅读
目的:研究带hTERT启动子的腺病毒通过干扰ERCC1基因表达而逆转卵巢癌顺铂耐药的作用。方法:以1、10、50、80感染复数(MOI)的重组腺病毒分别感染卵巢癌细胞株SKOV3、卵巢癌细胞顺铂耐药株SKOV3/DDP和脐静脉内皮细胞ECV304,同时以相同滴度的空载体腺病毒感染细胞,将感染前的细胞作为对照,以RT-PCR方法检测重组腺病毒转染细胞中ERCC1 mRNA表达,以Western blot方法检测重组腺病毒转染细胞中ERCC1蛋白表达,并以MTT法检测肿瘤细胞对顺铂化疗敏感性的影响。结果:(1)在SKOV3及SKOV3/DDP细胞中,不同剂量Ad-hTERT-ERCC1 shRNA转染组与转染前相比,ERCC1 mRNA及ERCC1蛋白表达水平下调,差异有统计学意义(P<0.01)。而在空载体腺病毒转染组及ECV304细胞中,ERCC1 mRNA及ERCC1蛋白表达水平无改变,差异无统计学意义(P>0.05);(2)重组腺病毒转染后,SKOV3/DDP细胞对顺铂的敏感性显著增加(P<0.01),并呈剂量依赖性。结论:带hTERT启动子的腺病毒能够通过干扰ER-CC1基因表达而逆转卵巢癌顺铂耐药,并且具有剂量依赖性。
OBJECTIVE: To study the role of adenovirus carrying hTERT promoter in reversing cisplatin-resistant ovarian cancer cells by interfering with ERCC1 gene expression. Methods: The ovarian cancer cell line SKOV3, SKOV3 / DDP ovarian cancer cells and ECV304 cells were infected with recombinant adenovirus at the MOI of 1, 10, 50 and 80 respectively. The same titer Transfected cells were used as control. The expression of ERCC1 mRNA was detected by RT-PCR in transfected cells. The expression of ERCC1 protein in recombinant adenovirus transfected cells was detected by Western blot The effect of tumor cells on chemosensitivity to cisplatin was detected by MTT assay. Results: (1) The expression of ERCC1 mRNA and ERCC1 protein in SKOV3 and SKOV3 / DDP cells transfected with different doses of Ad-hTERT-ERCC1 shRNA was significantly lower than that before transfection (P <0.01) ). However, the expression of ERCC1 mRNA and ERCC1 protein did not change in empty vector adenovirus transfection group and ECV304 cells (P> 0.05). (2) After transfected with recombinant adenovirus, SKOV3 / Cisplatin significantly increased the sensitivity (P <0.01), and in a dose-dependent manner. CONCLUSION: The adenovirus with hTERT promoter can reverse cisplatin-resistant ovarian cancer cells by interfering with ER-CC1 gene expression in a dose-dependent manner.