二甲双胍通过miR-342-3p/EGFR/Claudin 1抑制甲状腺乳头状癌细胞增殖、迁移和侵袭  

Metformin inhibits proliferation,migration and invasion of papillary thyroid carcinoma cells through miR-342-3p/EGFR/Claudin 1

作　　者：刘国力 张萌 李旭[1] LIU Guo-li;ZHANG Meng;LI Xu(Department of Pharmacy,The First Affiliated Hospital of China Medical University,Shenyang 110001;Department of General Surgery,Central Hospital Affiliated to Shenyang Medical College,Shenyang 110024,China)

机构地区：[1]中国医科大学附属第一医院药学部,辽宁沈阳110001 [2]沈阳医学院附属中心医院普外科,辽宁沈阳110024

出　　处：《解剖科学进展》2024年第4期413-417,共5页Progress of Anatomical Sciences

基　　金：沈阳医学院硕士研究生科技创新基金项目(Y20220531)。

摘　　要：目的探究二甲双胍对甲状腺乳头状癌细胞增殖、迁移和侵袭的影响及相关机制。方法不同浓度二甲双胍处理甲状腺乳头状癌TPC-1细胞48 h,CCK-8方法检测细胞增殖活性,Transwell实验检测细胞迁移和侵袭能力,RT-qPCR检测细胞miR-342-3p表达。将miR-342-3p inhibitor和阴性对照inhibitor转染TPC-1细胞,20 mmol/L二甲双胍处理48 h,CCK-8检测细胞增殖活性,Transwell实验检测细胞迁移和侵袭能力,Western blot检测细胞EGFR和Claudin 1蛋白表达。生物信息学分析miR-342-3p与EGFR mRNA的3′非翻译区的潜在结合位点,双荧光素酶报告基因实验验证miR-342-3p与EGFR mRNA结合,通过Western blot检测转染miR-342-3p mimics或miR-342-3p inhibitor对TPC-1细胞EGFR蛋白表达的影响。将EGFR过表达质粒和空载体转染TPC-1细胞,20 mmol/L二甲双胍处理48 h,Western blot检测细胞EGFR和Claudin 1蛋白表达。结果二甲双胍处理剂量依赖的抑制TPC-1细胞增殖、迁移和侵袭能力,上调细胞miR-342-3p表达。抑制miR-342-3p增加二甲双胍处理的TPC-1细胞增殖、迁移和侵袭能力。miR-342-3p与EGFR mRNA的3′非翻译区结合,过表达miR-342-3p下调TPC-1细胞EGFR蛋白表达且抑制miR-342-3p上调TPC-1细胞EGFR蛋白表达。抑制miR-342-3p或过表达EGFR增加二甲双胍处理的TPC-1细胞EGFR和Claudin 1蛋白表达。结论二甲双胍可能通过上调miR-342-3p抑制EGFR和Claudin 1表达抑制甲状腺乳头状癌细胞增殖、迁移和侵袭。Objective To explore the effects of metformin on the proliferation, migration and invasion of papillary thyroid carcinoma cells and the related mechanisms. Methods Papillary thyroid carcinoma TPC-1 cells were treated with different concentrations of metformin for 48 h. Cell proliferation activity was detected by CCK-8, cell migration and invasion ability was detected by Transwell assay, and the expression of miR-342-3p was detected by RT-qPCR.TPC-1 cells were transfected with miR-342-3p inhibitor and negative control inhibitor and treated with 20 mmol/L metformin for 48 h. Cell proliferation activity was detected by CCK-8 and cell migration and invasion ability was detected by Transwell assay. The expressions of EGFR and Claudin 1 protein were detected by Western blot. Bioinformatics was used to analyze the potential binding sites of the 3′ untranslated region of miR-342-3p and EGFR mRNA, and dual luciferase reporter assay was used to verify the binding of miR-342-3p to EGFR mRNA. The effect of transfection with miR-342-3p mimics or miR-342-3p inhibitor on expression of EGFR in TPC-1 cells was detected by Western blot. TPC-1 cells were transfected with EGFr-overexpressed plasmid and empty vector, and treated with 20 mmol/L metformin for 48 h. The expressions of EGFR and Claudin 1 protein were detected by Western blot. Results The dose-dependent treatment with metformin inhibited the proliferation, migration and invasion of TPC-1 cells and upregulated the expression of miR-342-3p. Inhibition of miR-342-3p increased the proliferation, migration and invasion ability of metformin-treated TPC-1 cells. miR-342-3p binds to the 3′ untranslated region of EGFR mRNA, and overexpression of miR-342-3p down-regulates EGFR protein expression in TPC-1 cells and inhibits miR-342-3p up-regulates expression of EGFR in TPC-1 cells. Inhibition of miR-342-3p or overexpression of EGFR increased the expressions of EGFR and Claudin 1 protein in metformin treated TPC-1 cells. Conclusion Metformin may inhibit the proliferation, migrati

关 键 词：二甲双胍 甲状腺乳头状癌 miR-342-3p EGFR Claudin 1 迁移 侵袭 

分 类 号：R736.1[医药卫生—肿瘤]