膜联蛋白A5低表达对人宫颈癌HeLa细胞迁移和侵袭的影响

发布时间：2018-05-03 04:34

  本文选题：膜联蛋白A + HeLa　； 参考：《解剖学报》2016年04期

【摘要】：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。
[Abstract]:Objective to investigate the effect of low expression of A5 A5 on migration and invasion of human cervical carcinoma HeLa cells. Methods the cells were divided into three groups: interference group, negative control group, transfection reagent control group and blank control group. SiRNA was transfected into HeLa cells by liposome transfection. After 72 hours of transfection, RT-PCR and Western blotting were used to detect the expression of ANXA5 in each group to identify the inhibition efficiency. The migration ability of HeLa cells in each group was detected by cell scratch assay. The invasion ability of HeLa cells in each group was detected by Transwell assay. The effects of low expression of ANXA5 on the expression of E-cadherin (E-cadherin) and matrix metalloproteinase-9 (MMP-9) and protein were detected by RT-PCR and Western blotting, respectively. Results the expression of ANXA5 protein and mRNA in the interference group was significantly lower than that in the negative control group (P 0.05). The expression of E-cadherin mRNA and protein in the interference group was significantly lower than that in the negative control group, and the expression of MMP-9mRNA and protein in the interference group was significantly higher than that in the negative control group. Conclusion siRNA targeting ANXA5 can effectively inhibit the expression of ANXA5, and the low expression of ANXA5 may promote the migration and invasion of HeLa cells by affecting the expression of E-cadherin and MMP-9.
【作者单位】： 承德医学院基础医学研究所;承德市266医院感染科;
【基金】：河北省高校省级重点学科项目(201304) 河北省科技厅课题(10276142) 河北省教育厅项目(Z2014025)
【分类号】：R737.33

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