摘要
目的探究miR-381调控高迁移率组蛋白1(HMGB1)对子宫内膜癌细胞增殖、侵袭和迁移能力的影响。方法采用qRT-PCR检测miR-381在子宫内膜癌细胞和正常子宫内膜细胞中的表达水平。转染miR-381mimic后,检测HEC1A细胞中miR-381和HMGB1的表达水平。生物信息学预测miR-381和HMGB1的靶向关系,并用荧光素酶报告实验加以验证。细胞分为HEC1A组、miR-381mimic组、pc-HMGB1组、mimic+pc-HMGB1组,采用CCK8法检测HEC1A细胞增殖能力,Transwell和划痕实验分别检测HEC1A细胞侵袭和迁移能力,蛋白印迹法检测HEC1A细胞中Ki67、PCNA、E-cadherin、N-cadherin、Vimentin、MMP-2和MMP-9的表达水平。结果 miR-381在正常子宫内膜细胞和癌细胞中存在表达差异。miR-381与HMGB1具有直接的靶向关系。与HEC1A组相比,miR-381组细胞增殖倍数、侵袭细胞数、划痕闭合率、Ki67、PCNA、N-cadherin、Vimentin、MMP-2和MMP-9的表达均降低(P<0.01),E-cadherin表达则增加(P<0.01),pc-HMGB1组上述指标则相反(P<0.01);与pc-HMGB1组相比,miR-381mimic+pc-HMGB1组细胞增殖倍数、侵袭细胞数、划痕闭合率、Ki67、PCNA、N-cadherin、Vimentin、MMP-2和MMP-9的表达均降低(P<0.01),E-cadherin表达增加(P<0.01)。结论 miR-381通过靶向下调HMGB1表达,抑制HEC1A细胞增殖、侵袭和迁移。
Objective To investigate the effect of miR-381 on the proliferation,invasion and migration of endometrial cancer cells by regulating high-mobility group box 1(HMGB1).Methods qRT-PCR was used to detect the mRNA level of miR-381 in endometrial cancer cells and normal endometrial cells.After transfection of miR-381 mimic,the expression levels of miR-381 and HMGB1 in HEC1 Acells were detected.Bioinformatics predicted the targeting relationship between miR-381 and HMGB1,and was verified by luciferase reporting experiments.Cells were divided into HEC1 Agroup,miR-381 mimic group,pc-HMGB1 group,mimic+pc-HMGB1 group.The proliferation ability of HEC1 Acells was detected by CCK8 method;the invasion and migration ability of HEC1 Acells were detected by Transwell and scratch test;and the expression levels of Ki67,PCNA,E-cadherin,N-cadherin,Vimentin,MMP-2 and MMP-9 in HEC1 Acells were detected by Western blotting.Results There were differences in the expression of miR-381 between normal endometrial cells and cancer cells.miR-381 has a direct targeting relationship with HMGB1.Compared with HEC1 Agroup,the expression of Ki67,PCNA,N-cadherin,Vimentin,MMP-2 and MMP-9 were decreased(P<0.01),while the expression of E-cadherin was increased(P<0.01),whereas those in pc-HMGB1 group were the opposite(P<0.01).Compared with pc-HMGB1 group,the expression of Ki67,PCNA,N-cadherin,Vimentin,MMP-2 and MMP-9 in miR-381+pc-HMGB1 group were decreased(P<0.01),while the expression of E-cadherin was increased(P<0.01).Conclusion MiR-381 inhibits cell proliferation,invasion and migration of HEC1 Acells by targeting downregulation of HMGB1 experssion.
作者
赵临潇
于东升
刘希
王雷
左博云
ZHAO Lin-xiao;YU Dong-sheng;LIU Xi(Department of Pathology,Xianyang Hospital,Yan'an University.Xiangyang,712000,China;Pathology Department of the First Affiliated Hospital of Xi'an Jiaotong University,Xi'an.Xi'an,710000,China)
出处
《中国实验诊断学》
2020年第6期989-995,共7页
Chinese Journal of Laboratory Diagnosis
基金
陕西省科技计划资助项目[编号:2017K14-G8(9)]。
关键词
miR-381
HMGB1
子宫内膜癌
增殖
侵袭
迁移
miR-381
high-mobility group box 1
endometrial cancer
proliferation
invasion
migration
