膜联蛋白A5低表达对人宫颈癌HeLa细胞迁移和侵袭的影响

目的探讨膜联蛋白A5(ANXA5)低表达对人宫颈癌HeLa细胞迁移和侵袭的影响。方法将细胞分为干扰组、阴性对照组、转染试剂对照组和空白对照组,采用脂质体转染法将siRNA转染干扰组HeLa细胞,在转染72h后采用RT-PCR和Western blotting检测各组ANXA5的表达以鉴定抑制效率;细胞划痕实验检测各组HeLa细胞的迁移能力;Transwell实验检测各组HeLa细胞的侵袭能力;RT-PCR和Western blotting分别检测ANXA5低表达对E-钙黏蛋白(E-cadherin)和基质金属蛋白酶(MMP)-9 mRNA及蛋白表达的影响。结果干扰组ANXA5蛋白及mRNA的表达均明显低于阴性对照组(P0.05);其表达均被显著抑制,干扰组细胞的迁移及侵袭能力比阴性对照组明显增强(P0.05),E-cadherin mRNA及蛋白的表达显著低于阴性对照组(P0.05);干扰组MMP-9mRNA及蛋白的表达均显著高于阴性对照组(P0.05)。结论靶向ANXA5的siRNA可有效抑制ANXA5的表达,且ANXA5低表达可能通过影响E-cadherin和MMP-9的表达来促进HeLa细胞的迁移和侵袭能力。

Effects of annexin A5 low expression on migration and invasion of HeLa cells

Objective To investigate the effects of annexin A5 (ANXA5) low expression on HeLa cell migration and invasion. Methods HeLa cells were divided into four groups, which were interference group, negative control group, and transfection reagent control group, and blank control group. RT-PCR and Western blotting were used to identify the inhibition efficiency at 72 hours after transfection. Cell scratch method and Transwell experiment were used to detect the ability of migration and invasion of HeLa cells in each group. E-cadherin and matrix metalloproteinase(MMP)-9 mRNA and protein were determined by RT-PCR and Western blotting respectively. Results HeLa cell ANXA5 expression on both mRNA and protein level was suppressed significantly(P<0.05). Compared with the negative control group, the abilities of migration and invasion of HeLa cells in the interference group were increased remarkably (P<0.05). The E-cadherin mRNA and protein expression of HeLa cells in interference group decreased while MMP-9 increased when compared with negative control group (P<0.05). Conclusion The siRNA which targets ANXA5 can highly suppress ANXA5 expression. ANXA5 knockdown promotes migration and invasion of HeLa cells via upregulation of E-cadherin and downregulation of MMP-9.