FMNL2基因小干扰RNA质粒转染人宫颈癌Hela、Siha细胞株Vimentin及E-cadherin的表达观察

摘要

Objective To investigate the expression of Vimentin and E-cadherin in human cervical carcinoma cell lines Hela and Siha transfected with Formin-like protein 2(FMNL2)gene small interference RNA (siRNA).Methods Both Hela cells and Siha cells were divided into the experimental group and control group, respectively.Cells in the experimental group were treated with 2 mL serum-free culture medium containing 100 nmol/L FMNL2-siRNA encapsulated by Lipofectamine 2000TM.Cells in the control group were treated with 2 mL serum-free culture medium.We used RT-PCR to detect the expression of FMNL2, Vimentin and E-cadherin mRNA in each group 72 h after transfection.Cellular immunohistochemistry was applied to detect the expression of FMNL2, Vimentin and E-Cadherin proteins in each group 72 h after transfection.Results The relative expression of FMNL2 mRNA and protein in both Hela cells and Siha cells of the experimental group 72 h after transfection was lower than that in the control group, respectively (P<0.05).The relative expression of Vimentin mRNA and protein in both Hela cells and Siha cells of the experimental group 72 h after transfection was lower than that in the control group, respectively, whereas the expression of E-cadherin mRNA and protein was higher than that in the control group (all P<0.05).Conclusions FMNL2 gene small interference RNA transfection may inhibit the expression of Vimentin and promote the expression of E-cadherin in Hela cells and Siha cells.FMNL2 may participate in the epithelial-mesenchymal transition of human cervical cancer.%目的 观察同源形成素样蛋白2(FMNL2)基因小干扰RNA 质粒转染人宫颈腺癌细胞株Hela、鳞癌细胞株Siha波形蛋白(Vimentin)及钙黏附蛋白E(E-cadherin)的表达的影响.方法 分别取Hela、Siha细胞各分为实验组、对照组,实验组中加入含有2 mL Lipofectamine 2000TM包裹的FMNL2-siRNA (100 nmol/L)的无血清培养基,对照组加2 mL的无血清培养基.采用RT-PCR法检测转染72 h时各组FMNL2、波形蛋白(Vimentin)及钙黏附蛋白E(E-cadherin) mRNA,采用细胞免疫化学染色法检测转染72 h时各组FMNL2、Vimentin及E-cadherin 蛋白.结果 转染72 h时Hela、Siha细胞实验组FMNL2 mRNA及蛋白的相对表达量均低于对照组,转染72 h时Hela、Siha细胞实验组Vimentin mRNA及蛋白的相对表达量均低于对照组,转染72 h时Hela、Siha细胞实验组 E-cadherin mRNA及蛋白的相对表达量mRNA相对表量均高于对照组(P均<0.05).结论 FMNL2基因小干扰RNA 质粒转染可抑制Hela、Siha细胞Vimentin 表达,促进 E-cadherin表达. FMNL2可能参与了宫颈癌EMT.