摘要
目的探究miR-198对宫颈癌细胞增殖、凋亡和侵袭的作用及其机制。方法用miR-198mimic转染HeLa细胞,qRT-PCR检测miR-198和丝裂原活化蛋白激酶1 (Mitogen-activated protein kinase1,MAPK1)的mRNA水平;荧光素酶实验检测miR-198和MAPK1的靶向关系;用pcDNA3.0-MAPK1(pc-MAPK1)和miR-198 mimic转染细胞,CCK-8检测细胞增殖活性,流式细胞术检测细胞凋亡,Transwell法检测细胞侵袭能力,Westernblot检测蛋白的表达。结果 miR-198mimic转染细胞后,miR-198表达水平明显升高,MAPK1mRNA表达水平明显降低;荧光素酶报告实验表明miR-198序列上存在MAPK1结合位点;miR-198过表达能显著降低宫颈癌细胞增殖倍数、诱导细胞凋亡,同时还能明显降低HeLa细胞侵袭能力;此外,miR-198 mimic能显著抑制MAPK1下游基因核糖体S6激酶2(Ribosomal S6 kinase 2, RSK2)、c-Myc和c-fos的蛋白表达;pc-MAPK1能明显减弱miR-198 mimic对HeLa细胞增殖、凋亡、侵袭及对MAPK1下游蛋白表达的调控作用。结论 miR-198过表达能通过靶向MAPK1抑制宫颈癌细胞的增殖和侵袭能力并诱导细胞凋亡。
Objective To investigate the effects and mechanisms of miR-198 on the proliferation,apoptosis and invasion of cervical cancer cells.Methods HeLa cells were transfected with miR-198 mimic and the mRNA levels between miR-198 and mitogen-activated protein kinase 1(MAPK1)were measured by qRTPCR.The relationship of miR-198 and MAPK1 was determined by luciferase reporter assay.pcDNA3.0-MAPK1(pc-MAPK1)and miR-198 mimic transfection was performed,and cell proliferation,apoptosis and invasion abilities were measured by CCK-8 assay,flow cytometry and Transwell assay,respectively.Western blot was performed for protein levels.Results The level of miR-198 was increased and the mRNA level of MAPK1 was decreased by miR-198 mimic.The result of luciferase reporter assay indicated that there was binding site of MAPK1 on miR-198.Overexpression of miR-198 significantly inhibited the proliferation and invasion,and induced the apoptosis of HeLa cells.In addition,miR-198 mimic down-regulated the expressions of ribosomal S6 kinase 2(RSK2),c-Myc and c-fos.pc-MAPK1 attenuated the effects of miR-198 mimic on cell proliferation,apoptosis,invasion and downstream proteins expression of MAPK1 in HeLa cells.Conclusion Overexpression of miR-198 inhibits the proliferation,invasion and induces the apoptosis of cervical cancer cells through targeting MAPK1.
作者
朱利红
段树鹏
秦海霞
张秀玲
赵淑珍
李少儒
王世进
ZHU Lihong;DUAN Shupeng;QIN Haixia;ZHANG Xiuling;ZHAO Shuzhen;LI Shaoru;WANG Shijin(Department of Obstetrics and Gynecology,The First Affiliated Hospital of Xinxiang Medical University,Xinxiang 453000,China;Department of Infection,The First Affiliated Hospital of Xinxiang Medical University,Xinxiang 453000,China)
出处
《肿瘤防治研究》
CAS
CSCD
2018年第12期959-964,共6页
Cancer Research on Prevention and Treatment
基金
河南省科技攻关项目(0124170680)