腺病毒介导的RNA干扰ERCC1基因表达对逆转卵巢癌顺铂耐药的效果

 目的 探讨腺病毒介导的RNA干扰ERCC1基因表达逆转卵巢癌顺铂耐药的效果。方法 采用直接感染法、台盼蓝活细胞计数、MTT、Hoechst染色法及流式细胞仪检测等方法,绘制顺铂耐药卵巢癌细胞株SKOV3/DDP细胞生长曲线及倍增时间,检测SKOV3/DDP细胞感染重组病毒前后其顺铂的IC₅₀值、细胞凋亡形态及细胞各周期的分布。结果 (1)顺铂浓度与SKOV3/DDP细胞抑制率呈线性正相关,相关系数r=0.9862(P<0.05),顺铂IC₅₀为(13.93±0.37)μg/ml,耐药指数为1.79。(2)重组腺病毒感染后干扰ERCC1基因表达,SKOV3/DDP细胞对顺铂的敏感性分别增加了11.1%、16.7%、26.4%、33.5%,呈剂量依懒性(r=0.9716,P<0.05),耐药指数降至1.19。(3)重组腺病毒联合顺铂作用后,SKOV3/DDP细胞株G₁期细胞比例增大,S期细胞比例增大,细胞凋亡率增高(P<0.05)。结论 干扰ERCC1基因表达可以通过诱导耐药肿瘤细胞SKOV3/DDP凋亡、调节细胞周期分布等途径逆转卵巢癌细胞的顺铂耐药。

Reversal of cisplatin resistance in ovarian cancer by adenovirus-mediated RNA interference with ERCC1 gene expression in vitro

Objective To investigate the effect of cisplatin resistance being reversed in ovarian cancer by adenovirus-mediated RNA interference with ERCC1 gene expression.Methods The cisplatin-resistant SKOV3/DDP cells were directly infected with the recombinant adenovirus, and trypan blue counting, MTT, Hoechst staining and flow cytometry were used in this study. The growth curve, doubling time, IC₅₀ value, apoptotic morphology and cell cycle scores of SKOV3/DDP cells before and after infection were detected.Results (1) The inhibition rate of SKOV3/DDP cells was positively correlated with the concentration of cisplatin. The correlation coefficient r=0.9862 (P<0.05). The IC₅₀ value of cisplatin was (13.93±0.37)μg/ml, and the resistance index was 1.79. (2) The sensitivity of SKOV₃/DDP cells to cisplatin increased by 11.1%, 16.7%, 26.4% and 33.5% respectively in a dose-dependent manner (r=0.9716, P<0.05) after being infected with the recombinant adenovirus, and the drug resistance index decreased to 1.19. (3) The proportion of G1 phase cells, S phase cells and apoptotic rate of SKOV3/DDP cells increased (P<0.05) after being infected with the recombinant adenovirus combined with cisplatin.Conclusions Interference with ERCC1 gene expressions can reverse cisplatin resistance in cisplatin-resistant ovarian cancer cell line SKOV3/DDP cells by inducing apoptosis and regulating cell cycle distribution.