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

发布时间：2018-04-09 07:04

本文选题：宫颈肿瘤　切入点：宫颈癌　出处：《山东医药》2017年19期

【摘要】：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。
[Abstract]:Objective to investigate the effect of small interference RNA plasmid of homologous morphogenetic protein 2FM-NL2 gene on the expression of Siha vimentin (Vimentin) and E-cadherin (E-cadherin) in human cervical adenocarcinoma cell line Hela, squamous-cell carcinoma cell line (Siha Vimentin) and calmodulin-like protein (E-cadherin).Methods Helaansiha cells were divided into experimental group and control group respectively. Serum-free medium containing 2 mL Lipofectamine 2000TM wrapped in FMNL2-siRNA(100 nmol / L was added in the experimental group and 2 mL serum-free medium was added in the control group.FMNL2, vimentin (Vimentin) and E-cadherin mRNAs were detected by RT-PCR method at 72 h after transfection, and FMNL2Vimentin and E-cadherin proteins were detected by cytoimmunocytochemical staining at 72 h after transfection.Results the relative expression of FMNL2 mRNA and protein in the experimental group was lower than that in the control group at 72 h after transfection, and the relative expression of Vimentin mRNA and protein in the experimental group was lower than that in the control group at 72 h after transfection.At 72 h after transfection, the relative expression of E-cadherin mRNA and protein in the experimental group was higher than that in the control group (P < 0.05).Conclusion transfection of small interfering RNA plasmid with FMNL2 gene can inhibit the expression of Vimentin in Helasiha cells and promote the expression of E-cadherin. FMNL2 may be involved in the expression of EMTs in cervical carcinoma.
【作者单位】：西南医科大学附属医院;
【基金】：四川省卫生厅科研基金资助项目(100243)
【分类号】：R737.33

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