摘要
目的:分析子痫前期(PE)患者胎盘及血清中长链非编码RNA(lncRNA)-kcnq1重叠转录物1(kcnq1ot1)表达情况,及其通过调控微小RNA(miRNA)19a对人脐静脉内皮细胞(HUVEC)功能的影响。方法:荧光实时定量聚合酶链式反应(qRT-PCR)检测15例PE晚孕患者(PE组)与15例正常晚孕患者(正常组)血清及胎盘组织中lncRNA-kcnq1ot1的表达;将HUVEC分为:过表达组(转染pcDNA3.1-kcnq1ot1)、抑制组(转染siRNA-kcnq1ot1)、对照组(未转染组)。qRT-PCR检测转染后三组HUVEC中miR-19a表达,蛋白免疫印迹(Western blot)检测三组HUVEC中CDKN2B、细胞周期蛋白D1(Cyclin D1)表达,管样形成实验检测三组HUVEC管腔数及总分支长度。结果:与正常组相比,PE组患者血清及胎盘组织中lncRNA-kcnq1ot1的表达降低(均P<0.05);与对照组相比,抑制组HUVEC中miR-19a在mRNA水平表达增高,CDKN2B、Cyclin D1在蛋白水平表达降低,且该组HUVEC管腔数及总分支长度降低(均P<0.05)。结论:lncRNA-kcnq1ot1在PE患者血清及胎盘组织中低表达,lncRNA-kcnq1ot1通过负性调控miR-19a表达,抑制HUVEC成管能力。
Objective:To analyze the expression of long-chain noncoding RNA(lncRNA)kcnq1 over lapping transcript 1(kcnq1ot1)in placenta and serum of patients with preeclampsia(PE),and its effect on the function of human umbilical vein endothelial cell(HUVEC)by regulating microRNA(miRNA)19a.Methods:The expression of lncRNA-kcnq1ot1 in serum and placenta of 15 late pregnancy patients with PE(PE group)and 15 normal late pregnancy patients(normal group)were detected by quantitative real time polymerase chain reaction(qRT-PCR).HUVEC cells were divided into overexpression group(transfected with pcDNA3.1-kcnq1ot1),inhibition group(transfected with siRNA-kcnq1ot1)and control group(non-transfected group).The expression of miR-19a in HUVEC cells was detected by qRT-PCR.Western blot was used to detect the expression of cyclin dependent kinase inhibitor 2B(CDKN2B)and cyclin D1 in HUVEC cells of the three groups.The lumen number and total branch length of HUVEC cells in the three groups were detected by tube like formation experiment.Results:Compared with the normal group,the expression of lncRNA-kcnq1ot1 in serum and placental tissue in PE group decreased(all P<0.05).Compared with the control group,the mRNA expression of miR-19a in HUVEC cells in the inhibition group increased,the protein expression of CDKN2B and Cyclin D1 decreased,and the lumen number and total branch length of HUVEC decreased(all P<0.05).Conclusion:LncRNA-kcnq1ot1 is low expressed in serum and placenta of patients with PE,and lncRNA-kcnq1ot1 inhibits the tube-forming ability of HUVEC by negatively regulating the expression of miR-19a.
作者
张媛
马媛
鲁杨
刘颖燕
周露秋
刘芳
ZHANG Yuan;MA Yuan;LU Yang;LIU Yingyan;ZHOU Luqiu;LIU Fang(Department of Obstetrics and Gynecology,Leshan People’s Hospital,Leshan 614000,China)
出处
《陕西医学杂志》
CAS
2022年第8期939-943,共5页
Shaanxi Medical Journal
基金
全国生殖医学中青年医生研究-基础项目(202206-09)
空军军医大学唐都医院科技创新发展基金资助项目(2017JCYJ002)。
