过表达mi R-129-5p通过靶向MAPK1抑制宫颈癌He La细胞恶性生物行为  

Over-expression of miR-129-5p inhibits malignant biological behaviors of cervical cancer HeLa cells by targeting MAPK1

作　　者：瞿小玲[1] 曾仪[2] 姚利[1] 蔡政[3] MAPK;ZENG Yi;YAO Li;CAI Zheng(Second Department of Obstetrics,Nanyang Central Hospital,Nanyang 473000,Henan,China;Department of Basic Medicine,Zhengzhou University,Zhengzhou 450000,Henan,China;Department of Oncology,Yunnan Provincial Hospital of Traditional Chinese Medicine,Kunming 650021,Yunnan,China)

机构地区：[1]南阳市中心医院产二科,河南南阳473000 [2]郑州大学基础医学部,河南郑州450000 [3]云南省中医医院肿瘤科,云南昆明650021

出　　处：《中国肿瘤生物治疗杂志》2019年第10期1075-1082,共8页Chinese Journal of Cancer Biotherapy

基　　金：河南省科技发展基金资助项目（No.132102310388）~~

摘　　要：目的:探讨miR-129-5p对宫颈癌HeLa细胞侵袭、迁移和EMT的作用及其机制。方法:选取宫颈癌HeLa细胞,利用生物信息学预测软件筛选miR-129-5p的靶基因,双荧光素酶报告基因验证miR-129-5p和MAPK1的靶向关系。将miR-129-5pmimic、miR-129-5pinhibitor和pcDNA-MAPK1单独或联合转染到HeLa细胞,用qPCR检测HeLa细胞中miR-129-5p和MAPK1的表达水平,用Transwell、划痕愈合实验分别检测HeLa细胞的侵袭、迁移能力,WB检测细胞中E-cadherin、N-cadherin、MAPK1、STAT3和Bcl-xL的表达。构建裸鼠HeLa细胞皮下移植瘤模型,观察miR-129-5p过表达对移植瘤生长的影响,WB检测移植瘤组织中EMT及MAPK1通路相关蛋白的表达。结果:miR-129-5p与MAPK1在3’UTR区存在结合位点,过表达miR-129-5p靶向抑制MAPK1(P<0.01)。与对照组相比,miR-129-5pmimic组侵袭细胞数目减少(P<0.01),划痕愈合率降低(均P<0.01);细胞中Ecadherin表达上调而N-cadherin、MAPK1、STAT3和Bcl-xL表达下调(均P<0.01);共转染MAPK1可逆转上述现象。成功建立裸鼠HeLa细胞移植瘤模型,与对照组相比,miR-128-3pmimic组肿瘤质量减轻(P<0.01);瘤组织中E-cadherin表达水平上调而N-cadherin、MAPK1、STAT3和Bcl-xL的表达下调(均P<0.01)。结论:过表达miR-129-5p通过靶向MAPK1抑制宫颈癌HeLa细胞的侵袭、迁移和EMT。Objective:To investigate the effects and mechanisms of miR-129-5p on invasion,migration and epithelial-mesenchymal transition(EMT)of cervical cancer HeLa cells.Methods:Cervical cancer HeLa cells were selected.The target gene of miR-129-5p was screened by bioinformatics prediction software,and the targeting relationship between miR-129-5p and MAPK1 was verified by dual luciferase reporter gene assay.HeLa cells were transfected with miR-129-5p mimic,miR-129-5p inhibitor and pcDNA-MAPK1 alone or in combination.The expressions of miR-129-5p and MAPK1 in HeLa cells were detected by qPCR;the invasion and migration ability of HeLa cells were detected by Transwell and scratch-healing experiments,respectively;and the expressions of E-cadherin,N-cadherin,MAPK1,STAT3 and Bcl-xL were detected by WB.The subcutaneous xenograft model of HeLa cells in nude mice was constructed to observe the effect of miR-129-5p over-expression on the growth of transplanted tumors.The expressions of EMT and MAPK1 pathwayrelated proteins in transplanted tumor tissues were detected by WB.Results:miR-129-5p could bind with the 3'UTR region of MAPK1,and over-expression of miR-129-5p targetedly inhibited the expression of MAPK1(P<0.01).Compared with the control group,the number of invasive cells in the miR-129-5p mimic group decreased(P<0.01),the scratch healing rate decreased(all P<0.01);The expression of E-cadherin was up-regulated,and the expressions of N-cadherin,MAPK1,STAT3 and Bcl-xL were down-regulated(all P<0.01),while co-transfection of MAPK1 reversed the above phenomenon.The nude mice HeLa cell xenograft model was successfully established.Compared with the control group,the tumor mass of the miR-128-3p mimic group was reduced;the expression of E-cadherin was up-regulated in tumor tissues,while the expressions of N-cadherin,MAPK1,STAT3 and Bcl-xL were down-regulated(all P<0.01).Conclusion:Over-expression of miR-129-5p inhibits invasion,migration and epithelial-mesenchymal transition of cervical cancer HeLa cells by targeting MAPK1.

关 键 词：宫颈癌 HELA细胞 miR-129-5p MAPK1 侵袭 迁移 上皮间质转化 

分 类 号：R737.33[医药卫生—肿瘤] R730.5[医药卫生—临床医学]