RNAi沉默cdc2基因对人卵巢癌顺铂耐药细胞株增殖与凋亡的影响

Effect of RNAi-silenced cdc2 gene on proliferation and apoptosis of cisplatin-resistant human ovarian cancer cell line

目的探讨利用RNAi沉默cdc2基因对人卵巢癌顺铂耐药细胞株A2780/DDP增殖、凋亡的影响,为卵巢癌顺铂耐药逆转提供一个可选择的靶点。方法针对cdc2基因的DNA设计具有特异性的SiRNA,经脂质体转染A2780/DDP,采用RT-PCR和免疫印迹法检测转染组cdc2基因表达是否抑制。用MTT法测定A2780/DDP细胞生长曲线,流式细胞仪检测细胞凋亡率及细胞周期改变。结果构建cdc2-SiRNA表达载体能够有效抑制A2780/DDP中cdc2基因的表达,差异有统计学意义(P<0.05)。RNAi沉默cdc2基因可抑制A2780/DDP细胞增殖,促进细胞凋亡,G2/M期细胞比例明显增加,差异有统计学意义(P<0.05)。结论通过RNAi抑制cdc2基因的表达,可抑制A2780/DDP细胞增殖,促进A2780/DDP细胞凋亡,为研究卵巢癌顺铂耐药逆转奠定一定基础。

Objective To explore the effect of RNA interference （RNAi） -silenced cdc2 gene on proliferation and apoptosis of eisplatin resistance of human ovarian cancer cell line A2780/DDP, provide a selectable target for reversion of cisplatin resistance of ovarian cancer. Methods Specific siRNA was established targeting to ode2 DNA, liposome was used to transfect A2780/DDP, RT-PCR and West- ern blotting were used to detect whether cde2 gene expression was inhibited in transfeetion group. MTI＂ was used to detect A2780/DDP cell growth curve, flow eytometry was used to detect apoptosis rate and the changes of cell cycle. Results cdc2- SiRNA expression vector effec- tively inhibited cde2 gene expression in A2780/DDP cells （P〈0. 05 ） . RNAi-silenced cdc2 gene inhibited proliferation of A2780/DDP cells, promoted cell apoptosis, the proportion of cells at G2/M phase increased, there were statistically significant differences （P〈0. 05 ） . Conclusion A2780/DDP ceils are inhibited by RNAi-silenced ode2 gene, cell proliferation is inhibited, cell apoptosis is promoted, which lays a foundation for studying reversion of cisplatin resistance of ovarian cancer.