长链非编码RNA XIST通过miR-186调控宫颈癌细胞侵袭迁移及凋亡的分子机制  

Molecular mechanisms of long chain non-coding RNA XIST regulating invasion,migration and apoptosis of cervical cancer cells by miR-186

作　　者：徐倩[1] 杨根岭 张波[3] 万斌 张瑜[5] XU Qian;YANG Gen-Ling;ZHANG Bo;WAN Bin;ZHANG Yu(College of Pharmacy,Chongqing Medical and Pharmaceutical College,Chongqing 400021,China)

机构地区：[1]重庆医药高等专科学校药学院,重庆401331 [2]重庆医科大学实验动物中心,重庆400016 [3]重庆市肿瘤医院针灸科,重庆400030 [4]重庆市中医院体检中心,重庆400021 [5]重庆医药高等专科学校中医学院,重庆401331

出　　处：《中国免疫学杂志》2020年第13期1600-1604,1609,共6页Chinese Journal of Immunology

基　　金：重庆市社会民生科技创新专项(No.cstc2015shmszxsydw-6)资助

摘　　要：目的:探讨长链非编码RNA(lncRNA)XIST通过miR-186调控宫颈癌细胞侵袭迁移及凋亡。方法:宫颈癌细胞中转染XIST shRNA,qRT-PCR检测XIST表达水平,流式细胞术检测细胞凋亡,Transwell小室检测细胞侵袭和迁移,Western blot检测Cleaved Caspase-3、MMP-2和MMP-9蛋白表达。生物信息学软件预测XIST和miR-186互补结合位点,荧光素酶报告载体鉴定。qRT-PCR检测XIST shRNA对miR-186表达影响。宫颈癌细胞中共转染XIST shRNA和miR-186 inhibitor,检测细胞凋亡、侵袭迁移和Cleaved Caspase-3、MMP-2、MMP-9蛋白表达水平,评估抑制miR-186对XIST shRNA影响宫颈癌细胞凋亡、侵袭和迁移的作用。结果:转染XIST shRNA可以下调宫颈癌细胞中XIST表达水平,促进细胞凋亡并抑制细胞侵袭和迁移,诱导Cleaved Caspase-3蛋白表达,减少MMP-2和MMP-9蛋白表达。XIST可靶向调控miR-186表达,XIST shRNA可提高miR-186表达。miR-186 inhibitor可明显逆转XIST shRNA对宫颈癌细胞miR-186表达促进、凋亡促进、侵袭迁移抑制和Cleaved Caspase-3、MMP-2、MMP-9蛋白表达影响。结论:下调XIST通过促进miR-186表达抑制宫颈癌细胞侵袭迁移并诱导细胞凋亡。Objective:To investigate role of long-chain non-coding RNA(lnc RNA)XIST in regulating invasion,migration and apoptosis of cervical cancer cells through miR-186.Methods:XIST shRNA was transfected into cervical cancer cells,expression of XIST was detected by qRT-PCR,cell apoptosis was detected by flow cytometry.Transwell chamber was used to by detect cell invasion and migration.Western blot was used to detect expressions of Cleaved Caspase-3,MMP-2 and MMP-9.Bioinformatics software predicted complementary binding sites of XIST and miR-186,and indentified by luciferase reporter vector,qRT-PCR was used to detect effect of XIST shRNA on expression of miR-186.XIST shRNA and miR-186inhibitor were co-transfected into cervical cancer cells,apoptosis,invasion,migration and expressions of Cleaved Caspase-3,MMP-2 and MMP-9 were detected,and evaluated effect of inhibiting miR-186 on XIST shRNA on apoptosis,invasion and migration of cervical cancer cells.Results:XIST shRNA can down-regulate expressions of XIST in cervical cancer cells,promote cell apoptosis and inhibit cell invasion and migration,induced expressions of Cleaved Caspase-3,MMP-2 and MMP-9.XIST can targeting regulate expression of miR-186,XIST shRNA can also increase expression of miR-186 in cells.miR-186 inhibitor can significantly reverse effects of XIST shRNA on expressions of miR-18,apoptosis,invasion and migration inhibition,and expression of Cleaved Caspase-3,MMP-2 and MMP-9 in cervical cancer cells.Conclusion:Down-regulation of XIST inhibits invasion and migration of cervical cancer cells and induces apoptosis by promoting expression of miR-186.

关 键 词：宫颈癌 XIST miR-186 侵袭 凋亡 

分 类 号：R737.33[医药卫生—肿瘤]