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目的:通过RNA干涉抑制Rab25在人卵巢上皮性癌的过表达,观察Rab25基因siRNA对卵巢癌细胞增殖及诱导凋亡的效果。方法:构建针对Rab25基因的siRNA重组质粒,稳定转染A2780细胞,通过RT-PCR检测Rab25的表达;MTT法观察细胞增殖,绘制细胞生长曲线;Hoechst33258荧光染色和透射电子显微镜观察细胞凋亡的形态学变化。结果:成功构建Rab25的siRNA,稳定转染靶细胞后可显著降低Rab25的表达,细胞生长速度减慢、增殖能力下降;荧光显微镜和电镜观察到细胞胞质浓缩、核凝聚、核碎裂和凋亡小体形成。结论:Rab25基因的siRNA能抑制细胞增殖,诱导细胞凋亡,为卵巢癌基因治疗开辟新途径。
OBJECTIVE: To investigate the effect of Rab25 siRNA on the proliferation and apoptosis of ovarian cancer cells by inhibiting the over-expression of Rab25 in human ovarian epithelial carcinoma by RNA interference. Methods: Rab25 recombinant plasmid was constructed and stably transfected into A2780 cells. The expression of Rab25 was detected by RT-PCR. The cell proliferation was observed by MTT assay and the cell growth curve was drawn. The morphology of apoptotic cells was observed by Hoechst33258 staining and transmission electron microscopy Learning changes. Results: Rab25 siRNA was successfully constructed and the expression of Rab25 was significantly decreased after stable transfection of target cells. The cell growth rate was slowed and the proliferation ability was decreased. The expression of Rab25 was observed by fluorescence microscope and electron microscope. Death body formation. Conclusion: siRNA of Rab25 gene can inhibit cell proliferation and induce apoptosis, opening up new ways for gene therapy of ovarian cancer.