siRNA抑制eIF4E基因的表达对卵巢癌A2780细胞生物学行为的影响

Effect of eIF4E Gene Inhibition by RNAi on the Biological Behaviors of Ovarian Cancer A2780 in Vitro

目的:探讨真核细胞翻译起始因子4E(eIF4E)基因在卵巢癌细胞生长增殖、迁移侵袭以及对顺铂敏感性中的作用。方法:采用eIF4E基因的特异shRNA表达质粒转染卵巢癌A2780细胞,通过siRNA抑制eIF4E的表达,采用real time-PCR和Western印迹法检测siRNA对eIF4EmRNA和蛋白表达水平的抑制作用,优化转染作用条件。进而分析eIF4E敲降后,A2780细胞生物学行为的变化。结果:特定的siRNA对eIF4E基因的表达有显著的抑制作用;eIF4E敲降后,与对照组比较,A2780细胞的增殖明显受到抑制(P<0.05),对顺铂的敏感性明显增强;细胞划痕实验和Transwell侵袭实验发现A2780细胞的运动侵袭能力显著下降(P<0.05);细胞周期分析发现A2780细胞S期细胞比例略有下降,G1期细胞比例略有上升,但无统计学意义(P>0.05)。结论:抑制eIF4E基因的表达,可以显著抑制卵巢癌细胞增殖、抑制细胞的迁移侵袭能力,增强细胞对顺铂敏感性。上述结果为卵巢癌的分子机制及顺铂敏感性的研究提供了新依据;也为卵巢癌的临床治疗提供了新靶点基因。

Objective：To study the role of eukaryotic translation initiation factor 4E（eIF4E）in the proliferation,migration,invasion and cisplatin resistance of human ovarian cancer（OC）cells.Methods：The shRNAs targeting eIF4 E were transfected into the human ovarian cancer cell A2780.The expression of eIF4 E were down-regulated by siRNA.Real-time PCR and Western blotting were used to detect the down-regulation of eIF4 EmRNA and protein and also used to optimize the conditions of the transfection.The roles of eIF4 Ein cell biologic behavior were analyzed with siRNA transfected cell.Results：The results revealed that eIF4 EsiRNA significantly down-regulated the expression of eIF4 Egene at both mRNA and protein levels.Down regulation of eIF4 Eexpression distinctly decreased the proliferation rates of the A2780 cells compared with the control group（P0.05）.And siRNA against eIF4 Esignificantly enhanced the therapeutic efficacy of cisplatin in A2780 cells.The cell wound healing assay and Transwell invasion assayshowed that the migration and invasion ability of A2780 cells significantly decreased by RNAi（P0.05）.The analysis of cell cycle revealed that the cellular proportion in S phase decreased slightly and the cellular proportion of Glphase increased slightly（P0.05）.Conclusion：Down regulation of eIF4 Eexpression distinctly decreased the proliferation,migration,and invasion ability,and enhanced the therapeutic efficacy of cisplatin in A2780 cells.These findings provide a new foundation for the study of the molecular mechanism and the cisplatin sensitivity in ovarian cancer.And these results indicate that siRNA against eIF4 Ehas the potential to become a new target for therapy of ovarian cancer.