靶向EGFR的干扰RNA对卵巢癌耐药细胞凋亡的影响

目的：探讨RNA干扰表皮生长因子受体（epidermal growth factor receptor，EGFR）的表达对多药耐药卵巢癌细胞株SKOV3/DDP凋亡的影响。方法：构建携带EGFR小发夹干扰RNA(small hairpin RNA, shRNA)的重组表达载体（pEGFRshRNA），脂质体法转染入SKOV3/DDP细胞，同时设未转染对照组和非特异性干扰质粒CtrlshRNA对照组。RTPCR和免疫细胞化学法检测转染后SKOV3/DDP细胞内EGFR mRNA和蛋白的表达；流式细胞仪分析EGFR沉默后SKOV3/DDP细胞周期和凋亡率的变化。结果：与转染对照质粒组相比，转染pEGFRshRNA组细胞EGFR mRNA和蛋白的表达明显受抑制(P<001)。流式细胞仪检测结果显示：顺铂作用24 h后，pEGFRshRNA转染组细胞周期分布发生明显改变，与对照组和CtrlsiRNA组相比，G0/G1期细胞比例和凋亡率明显增加（P<0.01），而S期细胞比例显著降低（P<0.01)；凋亡率显著升高。结论：靶向EGFR的干扰RNA可抑制SKOV3/DDP细胞中EGFR的表达，调节耐药细胞周期，促进耐药细胞凋亡。

Effect of EGFR targeted interference RNA on apoptosis of multidrug resistant ovarian cancer cells

Objective:To investigate the effect of RNA-mediated interference EGFR (epidermal growth factor receptor) expression on the apoptosis of muitidrug-resisitant ovarian cancer cell line SKOV3/DDP. Methods: Small hairpin RNA (shRNA) targeting EGFR was synthesized and recombinant plasmid containing pEGFR-shRNA was constructed, pEGFR-shRNA was tansfected into SKOV3/DDP cells by liposome system, untransfected cells and SKOV3/DDP cells tansfected with nonspecific-shRNA (Ctrl-shRNA) were used as control. Expression of EGFR mRNA and protein in SKOV3/DDP cells was examined by RT-PCR and immunocytochemistry after transfection, respectively. The apopototic rates and cell cy-cles of SKOV3/DDP cells were detected by flow cytometry. Results: Compared with Ctrl-shRNA-transfected cells, the ex-pression of EGFR mRNA and protein in pEGFR-shRNA-transfected SKOV3/DDP cells was significantly inhibited. Flow cytometry results showed that cell cycle distribution in pEGFR-shRNA-transfected SKOV3/DDP cells was dramatically changed, and the apoptosis rate was significantly increased after further treatment with cisplatin for 24 h. Conclusion: EGFR-targeted interference RNA can inhibit the expression of EGFR in SKOV3/DDP cells, thereby regulating the cell cy-cle and increasing apoptosis of multidrug-resistant SKOV3/DDP cells.