微小RNA-195靶向趋化因子5抑制滋养细胞增殖、迁移和侵袭及其机制研究  

MicroRNA-195 targeting chemokine 5 inhibits proliferation,metastasis and invasion of trophoblast cells and its mechanism

作　　者：程慧[1] 李妍雨 张蓓[1] 成杰[1] 张艳玲[1] Cheng Hui;Li Yanyu;Zhang Bei;Cheng Jie;Zhang Yanling(Department of Obstetrics and Gynecology,Xuzhou Central Hospital,Xuzhou 221009,Jiangsu Province,China)

机构地区：[1]徐州市中心医院妇产科,221009

出　　处：《中华妇幼临床医学杂志（电子版）》2022年第2期165-174,共10页Chinese Journal of Obstetrics & Gynecology and Pediatrics(Electronic Edition)

基　　金：江苏省卫计委科研计划(F201662)。

摘　　要：目的探讨微小RNA(miR)-195通过靶向调控趋化因子(CCL)5对胎盘滋养细胞增殖、侵袭的影响及其分子机制。方法采用随机数字表法,选取2018年8月至2019年8月在徐州市中心医院定期产前检查的70例孕妇,按照是否合并子痫前期(PE),将其分为PE组(n=40)和对照组(n=30)。选取2组受试者分娩时自胎盘母-胎界面组织中分离的滋养细胞株HTR-8/SVneo细胞为研究对象,针对PE组胎盘滋养层细胞分别转染miR-195及RNA阴性对照序列。根据转染结果,将PE组胎盘滋养层细胞分为miR-195 mimics亚组、miR-NC亚组、miR-195+pcDNA亚组、miR-195+pcDNA-CCL5亚组。采用实时荧光定量聚合酶链式反应(RT-PCR)检测研究组与对照组胎盘组织中miR-195、CCL5 mRNA相对表达水平;MTT实验检测各亚组滋养细胞增殖能力,Transwell侵袭实验观察细胞侵袭能力,细胞划痕实验观察各亚组滋养细胞迁移能力,Western blotting法检测CCL5、PI3K及AKT蛋白相对表达水平;Pearson相关性分析对miR-195及CCL5表达相关性进行分析;荧光素酶实验验证miR-195与CCL5的靶向关系。本研究遵循的程序符合徐州市中心医院伦理委员会规定,通过该伦理委员会审查,并获得批准(审批文号:XZXY-LI-20181215-031)。所有受试者均签署临床研究知情同意书。结果①PE组和对照组孕妇年龄、孕龄等一般临床资料比较,差异无统计学意义(P>0.05)。2组孕妇血压及尿蛋白水平比较,差异有统计学意义(P<0.05)。②PE组患者胎盘组织中miR-195 mRNA相对表达水平高于对照组(t=10.932,P<0.001);PE组患者胎盘组织中CCL5 mRNA、蛋白相对表达水平低于对照组(t=11.125、13.253,P<0.001)。③miR-195 mimics亚组滋养细胞中miR-195相对表达水平高于miR-NC亚组(P<0.001),miR-195 mimics亚组第1、2、3、4天细胞活力低于miR-NC亚组,并且差异均有统计学意义(P<0.05)。④细胞培养72 h后,miR-195 mimics亚组侵袭细胞数、细胞迁移率均低于miR-NC亚组,并且�Objective To investigate effects of microRNA(miR)-195 on the proliferation and invasion of trophoblast cells by targeting chemokine(CCL)5 and its molecular mechanism.Methods Forty pre-eclampsia(PE)pregnant women(PE group)and 30 healthy pregnant women(control group)were selected by randon number table method.The trophoblast cell line HTR-8/SVneo was transfected with miR-195 and RNA-negative control sequences,respectively.According to the transfer results,placental trophoblast cells in PE group were divided into miR-195 mimics subgroup,miR-NC subgroup,miR-195+pcDNA subgroup,miR-195+pcDNA-CCL5 subgroup.Real-time polymerase chain reaction(RT-PCR)was used to detect the expression level of miR-195,CCL5 mRNA.The cell proliferation ability of each group was detected by MTT.The cell invasion ability was detected by Transwell invasion experiment.The ability of cell migration in each group was observed by cell scratch test.The expression of CCL5,PI3K and AKT protein in each group were detected by Western blotting.The correlation between the expressions of miR-195 and CCL5 was analyzed by Pearson correlation analysis.Luciferase experiment was used to verify the targeting relationship between miR-195 and CCL5.The procedure followed in this study met the standards formulated by the Ethics Review Committee of Xuzhou Central Hospital and has been approved by it(Approval No.KS2204).Informed consent was obtained from each participant.Results①There was no significant difference between PE group and control group in general clinical data such as age,gestational age(P>0.05).The blood pressure and urine protein levels of pregnant women in two groups were significantly different(P<0.05).②The relative expression level of miR-195 mRNA in placental tissue of PE group was higher than that of control group(t=10.932,P<0.001),and the relative expression of CCL5 mRNA and protein in placental tissue of PE group were lower than those of control group(t=11.125,13.253;P<0.001).③The level of miR-195 in miR-195 mimics subgroup was higher than

关 键 词：微RNAS 微小RNA-195 趋化因子CCL5 滋养细胞 细胞增殖 侵袭 先兆子痫 

分 类 号：R73[医药卫生—肿瘤]