长链非编码RNA X失活特异转录物调控miR-340-5p对卵巢癌细胞上皮间质转化的影响  

Influence of long non-coding RNA XIST on epithelial-mesenchymal transition in ovarian cancer cells by regulating miR-340-5p

作　　者：姜平[1] 郭哲[1] 梁殿迅[1] 杨喜科[1] 付小玲[1] 李和丽 JIANG Ping;GUO Zhe;LIANG Dianxun;YANG Xike;FU Xiaoling;LI Heli(He'nan Nanyang Central Hospital,Nanyang 473000,China)

机构地区：[1]河南省南阳市中心医院,河南南阳473000

出　　处：《实用医学杂志》2023年第1期41-47,共7页The Journal of Practical Medicine

基　　金：河南省医学科技攻关项目(编号:172103210067;202003210381)。

摘　　要：目的探讨长链非编码RNA X失活特异转录物(lncRNA XIST)通过调控miR-340-5p对卵巢癌细胞上皮间质转化(EMT)的影响。方法实时荧光定量PCR(RT-qPCR)检测卵巢癌组织和细胞系中lncRNA XIST和miR-340-5p表达水平。A2780细胞分为Control组、si-NC组、si-lncRNA XIST组、si-lncRNA XIST+NC inhibitor组、si-lncRNA XIST+miR-340-5p inhibitor组。双荧光素酶实验证实lncRNA XIST和miR-340-5p的调控关系;划痕实验和Transwell法检测迁移和侵袭能力;RT-qPCR、Western blot和免疫荧光染色分析EMT标志蛋白(E-cadherin、N-cadherin、Vimentin)的mRNA和蛋白表达。结果在卵巢癌组织和细胞系中,lncRNA XIST高表达,miR-340-5p低表达(P<0.05)。选择lncRNA XIST表达最高的A2780细胞作为转染对象。lncRNA XIST能够直接靶向下调miR-340-5p表达(P<0.05)。转染si-lncRNA XIST后,lncRNA XIST水平、划痕愈合率、侵袭细胞数和N-cadherin、Vimentin表达降低,E-cadherin表达升高(P<0.05);共转染si-lncRNA XIST和miR-340-5p inhibitor后上述指标均得到逆转(P<0.05)。结论lncRNA XIST通过下调miR-340-5p促进卵巢癌细胞EMT。Objective To investigate the influence of long non-coding RNA X-inactive specific transcript(lncRNA XIST)on epithelial-mesenchymal transition(EMT)of ovarian cancer cells by regulating miR-340-5p.Methods Real-time quantitative PCR(RT-qPCR)was performed to determine expression levels of lncRNA XIST and miR-340-5p in ovarian cancer tissues and cell lines.A2780 cells were divided into a control group,si-NC group,si-lncRNA XIST group,si-lncRNA XIST+NC inhibitor group,and si-lncRNA XIST+miR-340-5p inhibitor group.Dual-luciferase reporter assay was performed to confirm the regulatory relationship between lncRNA XIST and miR-340-5p;scratch test and Transwell assay were performed to detect the migration and invasion abilities ofthe cells.The mRNA and protein expressions of EMT marker proteins(E-cadherin,N-cadherinand Vimentin)were analyzed by RT-qPCR,Western blot andimmunofluorescence staining.Results In ovarian cancer tissues and cell lines,lncRNA XIST was highly expressed,and miR-340-5p was in low expression(P<0.05).The A2780 cells with the highest expression of lncRNA XIST were selected as a transfection target.lnc RNA XIST was able to directly down-regulate the expression of mil-340-5p(P<0.05).After transfection of si-lncRNA XIST,the level of lncRNA XIST,scratch healing rate,number of invasive cells,and expressions of N-cadherin and Vimentin were decreased,whereasexpression of E-cadherin was increased;after co-transfection of si-lncRNA XIST with miR-340-5p inhibitor,the above indexes were reversed(P<0.05).Conclusions LncRNA XIST accelerates EMT in ovarian cancer cells by down-regulating miR-340-5p.

关 键 词：长链非编码RNA X失活特异转录物 miR-340-5p 卵巢癌 上皮间质转化 

分 类 号：R737.31[医药卫生—肿瘤]