过/降表达miR-4746的食管鳞癌细胞增殖、迁移、侵袭能力及TGF-β/Smad通路蛋白表达观察  

Proliferation,migration and invasion and protein expression of TGF-β/Smad pathway of esophageal squamous cell carcinoma cells with over-/down-expression of miR-4746

作　　者：李雨珊 张仁静 侯维 蹇顺海[1,3] 向月 何欣蓉[1,3] LI Yushan;ZHANG Renjing;HOU Wei;JIAN Shunhai;XIANG Yue;HE Xinrong(Department of Pathology,The Affiliated Hospital of North Sichuan Medical College,Nanchong 637000,China;不详)

机构地区：[1]川北医学院附属医院病理科,四川南充637000 [2]南充市中心医院检验科 [3]川北医学院病理教研室

出　　处：《山东医药》2022年第5期30-34,共5页Shandong Medical Journal

基　　金：川北医学院科研发展计划项目(CBY21-QA23)。

摘　　要：目的观察过/降表达miR-4746的食管鳞癌细胞增殖、迁移、侵袭能力及转化生长因子β(TGF-β)/Smad通路蛋白的表达。方法选取人食管鳞癌细胞ECA-109、TE-1和正常食管上皮细胞HEEC,采用实时荧光定量PCR法检测各细胞中miR-4746的表达水平。取miR-4746表达最高的食管鳞癌细胞分为miR-4746 mimic组、miR-4746 inhibitor组及对照组,分别转染miR-4746过表达、miR-4746抑制及对照质粒。采用CCK-8法观察细胞增殖能力,划痕实验观察细胞迁移能力,Transwell实验观察细胞侵袭能力。通过实时荧光定量PCR法和Western blotting法分别检测TGF-β/Smad通路相关分子TGF-β_(1)、TGF-βRⅡ、Smad4的mRNA和蛋白相对表达量。结果miR-4746在ECA-109、TE-1细胞中的相对表达量均高于HEEC(P均<0.01),其中TE-1细胞的miR-4746表达量最高。各时点细胞吸光度值、细胞迁移距离、穿膜细胞数比较,miR-4746 mimic组>对照组>miR-4746 inhibitor组(P均<0.05)。TGF-β_(1)mRNA及蛋白表达量miR-4746 mimic组>对照组>miR-4746 inhibitor组,TGF-βRⅡmRNA、Smad4 mRNA及蛋白表达量miR-4746inhibitor组>对照组>miR-4746 mimic组(P均<0.05),各组TGF-βRⅡ蛋白表达量比较差异无统计学意义。结论miR-4746可促进食管鳞癌细胞的增殖、迁移及侵袭,其作用可能通过调控Smad4低表达,抑制TGF-β/Smad通路实现。Objective After over-expression or down-expression of miR-4746 in the esophageal squamous carcinoma(ESCC)cells,the proliferation,migration,invasion and protein expression of transforming growth factorβ(TGF-β)/Smad pathway were observed.Methods The expression levels of microRNA-4746(miR-4746)in ESCC cells ECA-109,TE-1 and normal esophageal epithelial cells HEEC were detected by real-time quantitative PCR.The ESCC cells with the highest expression of miR-4746 were divided into the miR-4746 mimic group,miR-4746 inhibitor group,and control group,which were transfected with miR-4746 overexpression,miR-4746 inhibition,and control plasmid,respectively.CCK-8 was used to observe cell proliferation,Scratch test to observe cell migration,and Transwell test to observe cell invasion.The relative mRNA and protein expression levels of TGF-β_(1),TGF-βRⅡ,and Smad4 in TGF-β/Smad pathway were detected by real-time quantitative PCR and Western blotting,respectively.Results The relative expression of miR-4746 in ECA-109 and TE-1 cells was higher than that in HEEC(all P<0.01),and the expression of miR-4746 in TE-1 cells was the highest.A value,cell migration distance,and the number of transmembrane cells were higher in the miR-4746 mimic group than in the control group,and the miR-4746 mimic group had the lowest number(all P<0.05).TGF-β1 mRNA and protein expression levels were higher in the miR-4746 mimic group than in the control group,and were the lowest in the miR-4746 inhibitor group.TGF-βRⅡmRNA,Smad4 mRNA and protein expression levels were higher in the miR-4746 inhibitor group than in the control group,and were lowest in the miR-4746 inhibitor group(all P<0.05).There was no significant difference in the TGF-βRⅡprotein expression between groups.Conclusion MiR-4746 can promote the proliferation,migration and invasion of ESCC cells by regulating the low expression of Smad4 and inhibiting the TGF-β/Smad pathway.

关 键 词：miR-4746 食管鳞癌 细胞增殖 细胞迁移 细胞侵袭 转化生长因子Β SMAD4 

分 类 号：R735.1[医药卫生—肿瘤]