LINC00511通过靶向对滋养层细胞迁移侵袭和上皮间质转化的影响  

LINC00511 regulates trophoblast cell migration,invasion and epithelial mesenchymal transformation by targeting miR-16-5p

作　　者：张芹 卫兵[1] 王文艳[1] ZHANG Qin;WEI Bing;WANG Wen-yan(Department of Obstetrics and Gynecology,The Second Affiliated Hospital of Anhui Medical University,Hefei 230022,Anhui,China)

机构地区：[1]安徽医科大学第二附属医院妇产科,合肥230022

出　　处：《医学研究生学报》2022年第2期118-124,共7页Journal of Medical Postgraduates

基　　金：国家自然科学基金(81100412);中国博士后科学基金面上项目(2016M592038);高校优秀青年人才支持计划重点项目(gxyqZD2016060)。

摘　　要：目的 LINC00511对滋养层细胞的影响及其机制是否与miR-16-5p有关尚未完全明确。文中旨在探讨LINC00511对人绒毛膜滋养层细胞HTR-8/Svneo迁移侵袭和上皮间质转化(EMT)的影响及其分子机制。方法 选取2016年1月至2019年1月安徽医科大学第二附属医院妇产科60例剖宫产产妇的的胎盘组织。检测30个子痫前期(PE)胎盘和30个正常胎盘组织中LINC00511和miR-16-5p的表达水平;体外培养HTR-8/Svneo细胞,将其分为pcDNA组、pcDNA-LINC00511组、anti-miR-NC组、anti-miR-16-5p组、pcDNA-LINC00511+miR-NC组、pcDNA-LINC00511+miR-16-5p组;RT-qPCR检测LINC00511和miR-16-5p的表达水平;Transwell实验检测细胞的迁移侵袭能力;Western blot检测相关蛋白的表达;双荧光素酶报告实验检测LINC00511和miR-16-5p的靶向关系。结果 PE胎盘组织LINC00511表达水平(0.45±0.03)较正常胎盘组织(1.01±0.05)显著降低(P<0.05),miR-16-5p的表达水平(2.93±0.15)较正常胎盘组织(0.99±0.07)显著升高(P<0.05)。pcDNA-LINC00511组LINC00511表达水平较pcDNA组明显升高,迁移细胞数、侵袭细胞数、MMP-2、MMP-9、Vimentin、N-cadherin和Twist1的表达水平显著升高,E-cadherin的表达水平显著降低(P<0.05)。pcDNA-LINC00511组miR-16-5p表达水平较pcDNA组明显降低(P<0.05);si-LINC00511组miR-16-5p表达水平较si-NC组明显升高(P<0.05)。与anti-miR-NC组比较,anti-miR-16-5p组miR-16-5p的表达水平显著降低,迁移细胞数、侵袭细胞数、MMP-2、MMP-9、Vimentin、N-cadherin和Twist1的表达水平显著升高,E-cadherin的表达水平显著降低(P<0.05)。与pcDNA-LINC00511+miR-NC组比较,pcDNA-LINC00511+miR-16-5p组miR-16-5p的表达水平显著升高,迁移细胞数、侵袭细胞数、MMP-2、MMP-9、Vimentin、N-cadherin和Twist1的表达水平显著降低,E-cadherin的表达水平显著升高(P<0.05)。结论 LINC00511对miR-16-5p表达水平起到靶向下调作用,通过该途径对HTR-8/Svneo细胞迁移侵袭、EMT过程发挥一定的促进作用。Objective Both LINC00511 and miR-16-5 p can affect the biological behavior of trophoblast cells. However, the role of LINC00511 on trophoblast cells and its mechanism underlying to miR-16-5 p have not been completely clarified. Methods The expression levels of LINC00511 and miR-16-5 p in 30 preeclampsia(PE) placentas and 30 normal placentas were detected. HTR-8/Svneo cells were cultured in vitro and divided into pcDNA group, pcDNA-LINC00511 group, anti-miR-NC group, anti-miR-16-5 p group, pcDNA-LINC00511+miR-NC group, pcDNA-LINC00511+miR-16-5 p group. RT-qPCR was used to detect the expression levels of LINC00511 and miR-16-5 p. The ability of cell migration and invasion were detected by Transwell assays.. Western blot was used to detect the expression of related proteins. Dual luciferase reporter assay was used to detect the targeting relationship between LINC00511 and miR-16-5 p. Results Compared with normal placental tissues, the expression levels of LINC00511 and miR-16-5 p in PE placental tissues were significantly decreased and increased, respectively(P<0.05). Upregulating the expression of LINC00511 significantly promoted the migration, invasion and EMT of HTR-8/Svneo cells. The expression levels of MMP-2, MMP-9, Vimentin, N-cadherin and Twist1 were increased, while the expression levels of E-cadherin was decreased(P<0.05). LINC00511 targeted miR-16-5 p. Inhibition of miR-16-5 p expression significantly promoted the migration, invasion and EMT of HTR-8/Svneo cells. The expression levels of MMP-2, MMP-9, Vimentin, N-cadherin and Twist1 were increased, while the expression levels of E-cadherin was decreased(P<0.05). After the up-regulation of LINC00511 expression, EMT and migration invasion of HTR-8/SVneo cells were affected, while miR-16-5 p overexpression reversed the effect(P<0.05). Conclusion LINC00511 plays a targeted down-regulation role in miR-16-5 p expression, and plays a certain role in promoting the migration and invasion of HTR-8/SVneo cells and the EMT process through this pathway.

关 键 词：LINC00511 miR-16-5p 子痫前期 迁移 侵袭 上皮间质转化 

分 类 号：R714[医药卫生—妇产科学]