miR-490-3p调控SW1990胰腺癌细胞上皮间充质转化  

miR-490-3p Suppresses EMT of SW1990 Pancreatic Cancer Cells

作　　者：李京辉 朱明 曲海 李秋宇 吴玉娟 杨贺英 王郁竹 李妍平 LI Jing-hui;ZHU Ming;QU Hain;LI Qiu-yu;WU Yu-juan;YANG He-ying;WANG Yu-zhu;LI Yan-ping(Dept.of Critical Care Medicine,Yan'an Hospital of Kunming,Kunming Yunnan 650051,China;Dept.of Neurology,Yan'an Hospital of Kunming,Kunming Yunnan 650051,China)

机构地区：[1]昆明市延安医院重症医学科,云南昆明650051 [2]昆明市延安医院神经内科,云南昆明650051

出　　处：《昆明医科大学学报》2021年第3期10-17,共8页Journal of Kunming Medical University

基　　金：昆明市卫生健康委员会卫生科研基金资助项目(2020-21-01-011);昆明市延安医院院内基金资助项目(yyky019-040)。

摘　　要：目的研究miR-490-3p在分子水平上通过HMGA2蛋白对SW1990细胞上皮间充质转化的影响。方法通过实时荧光定量PCR法(RT-qPCR)检测正常胰腺导管上皮细胞HPDE和人胰腺癌细胞SW1990中miR-490-3p的表达。通过转染质粒[miR-490-3p阻遏物(inhibitor)和miR-490-3p模拟物(mimic)以及各自的阴性对照质粒]调控miR-490-3p在SW1990细胞中的表达并通过RT-qPCR法验证转染效率。转染48 h后,通过CCK8检测、划痕法、Transwell小室检测,流式细胞仪检测等分别检测细胞增殖、凋亡、迁移和侵袭等恶性生物学特征的影响。通过Western blot检测细胞中上皮间充质转化相关蛋白E-cadherin和N-cadherin的蛋白表达水平。同时,用数据库预测miR-490-3p及其靶基因HMGA2的靶向关系,并通过双荧光素酶报告基因实验进行验证,通过Western blot检测细胞中HMGA2的蛋白表达水平。结果(1)与HPDE正常细胞相比,SW1990癌细胞中miR-490-3p表达水平明显降低(P=0.215);(2)miR-490-3p上调后,SW1990细胞的凋亡率显著高于对照组(P<0.0001),而细胞增殖、迁移和侵袭能力显著低于对照组(P<0.0001)。miR-490-3p下调后,SW1990细胞的凋亡率显著低于对照组(P<0.0001),而细胞增殖、迁移和侵袭能力显著高于对照组(P<0.0001);(3)miR-490-3p上调后,SW1990细胞中的N-cadherin表达量显著高于对照组(P<0.0001),而E-cadherin的表达水平显著低于对照组(P<0.0001);miR-490-3p下调后,SW1990细胞中的N-cadherin表达量显著低于对照组(P<0.0001),而E-cadherin的表达水平显著高于对照组(P<0.0001);(4)HMGA2是miR-490-3p的靶向基因。结论miR-490-3p可通过靶向HMGA2抑制SW1990胰腺癌细胞EMT,从而影响胰腺癌的的发生发展进程,揭示HMGA2和miR-490-3p可能为胰腺癌诊断和治疗的靶点。Objective To elucidate the role of miR-490-3p in regulating HMGA2 expression and affecting epithelial-mesenchymal transition(EMT)of SW1990 cells.Methods miR-490-3p expression was measured in HPDE and SW1990 cell lines by Real-time PCR.SW1990 cell lines were cultured in vitro and divided into four groups,including inhibitor-Negative control(inhibitor-NC),miR-490-3p inhibitor,mimic-Negative control(mimic-NC)and miR-490-3p mimic group,and the transfection efficiency was verified by RT-qPCR.After 48 h of transfection,CCK8 assay,wound healing assay,Transwell invasion assay andflow cytometric were used to detect the effects of malignant biological characteristics such as proliferation,migration,invasion and apoptosis of SW1990 cell linerespectively.Western blot was used to detect the expression of E-cadherin and N-cadherin.Meanwhile,database predicted the target genes of miR-490-3p,and dual luciferase assay confirmed the targeted relationship between miR-490-3p and HMGA2,western blot was used to detect the expression of HMGA2.Results 1)The expression of miR-490-3p was downregulated in SW1990 cells(P=0.215).2)Overexpression of miR-490-3p significantly inhibited the proliferation(P<0.0001),migration(P<0.0001)and invasion(P<0.0001)of SW1990 cell line and prompted apoptosis(P<0.0001);In contrast,downregulation of miR-490-3p significantly promoted the proliferation(P<0.0001),migration(P<0.0001)and invasion(P<0.0001)of SW1990 cell line and inhibited apoptosis(P<0.0001).3)Upregulation of miR-490-3p significantly decreased the expression of N-cadherin(P<0.0001)and increased E-cadherin(P<0.0001)expression in SW1990 cell line.In contrast,downregulation of miR-490-3p significantly upregulated(P<0.0001)the expression of N-cadherin,decreased E-cadherin(P<0.0001)expression.4)HMGA2 was a target gene of miR-490-3p.Conclusion These findings indicate that miR-490-3p acts as a tumor suppressor during the process of EMT through targeting HMGA2,suggesting miR-490-3p is a potential new diagnostic and therapeutic target for the treatment of

关 键 词：胰腺癌 上皮间充质转化 miR-490-3p HMGA2 

分 类 号：R737.31[医药卫生—肿瘤]