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目的研究调控Rab26蛋白表达对宫颈癌HeLa细胞凋亡及迁移的影响。方法常规培养HeLa细胞,分别将含有Rab26 cDNA全长的真核表达载体(过表达组)、含有Rab26 siRNA序列的真核表达载体(siRNA组)瞬时转染HeLa细胞,同时设立正常细胞组及空载体转染组作为对照。转染48 h后,分别用RT-PCR和Western blot检测各组细胞中Rab26的mRNA及蛋白表达水平;采用流式细胞仪(FCM)检测各组细胞的凋亡率;同时采用划痕实验检测各组细胞迁移速度。结果与正常细胞组及空载体转染组相比,过表达组HeLa细胞中Rab26 mRNA及蛋白表达水平显著上调(P<0.05),而siRNA组细胞中Rab26 mRNA及蛋白表达水平显著下调(P<0.05),过表达组HeLa细胞凋亡率增高并抑制细胞迁移速度(P<0.01),siRNA组则促进细胞迁移(P<0.05)。结论 Rab26与HeLa细胞的凋亡及细胞迁移有关,故调控Rab26的表达有望成为治疗宫颈癌的新靶点。
Objective To study the effect of Rab26 protein expression on apoptosis and migration of cervical cancer HeLa cells. Methods HeLa cells were cultured routinely. HeLa cells were transiently transfected with eukaryotic expression vector containing Rab26 cDNA (overexpression group) and eukaryotic expression vector (siRNA group) containing Rab26 siRNA sequence, and normal cells and empty cells Vector transfected group served as control. The mRNA and protein expression of Rab26 in each group were detected by RT-PCR and Western blot respectively 48 h after transfection. The apoptosis rate of each group was detected by flow cytometry (FCM) The rate of cell migration in each group. Results The expression of Rab26 mRNA and protein in HeLa cells was significantly upregulated (P <0.05), while the expression of Rab26 mRNA and protein was significantly down - regulated in HeLa cells compared with normal cells and empty vector transfected cells (P < 0.05). The overexpression of HeLa increased the apoptotic rate and inhibited the migration of HeLa cells (P <0.01), while siRNA decreased the migration of HeLa cells (P <0.05). Conclusion Rab26 is related to the apoptosis and cell migration of HeLa cells. Therefore, the regulation of Rab26 may be a new target for the treatment of cervical cancer.