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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 investigate the effects of cdc2 gene silencing on the proliferation and apoptosis of cisplatin-resistant human ovarian cancer cell line A2780 / DDP by RNA interference and to provide a selectable target for the reversal of cisplatin-resistant ovarian cancer. Methods siRNA specific for cdc2 gene was designed and transfected into A2780 / DDP by liposome. The expression of cdc2 gene was detected by RT-PCR and Western blotting. The growth curve of A2780 / DDP cells was determined by MTT assay. The apoptosis rate and cell cycle were detected by flow cytometry. Results The construction of cdc2-SiRNA expression vector can effectively inhibit the expression of cdc2 gene in A2780 / DDP, with statistical significance (P <0.05). The silencing of cdc2 gene by RNAi could inhibit the proliferation of A2780 / DDP cells and promote the apoptosis of cells. The proportion of cells in G2 / M phase was significantly increased (P <0.05). Conclusion The inhibition of cdc2 gene expression by RNAi can inhibit the proliferation of A2780 / DDP cells and promote the apoptosis of A2780 / DDP cells, which will lay a foundation for the study of reversal of cisplatin resistance in ovarian cancer.