摘要
目的:探究静息巯基氧化酶-1(quiescin sulfhydryl oxidase 1,QSOX1)在子痫前期(preeclampsia,PE)胎盘中的表达情况,QSOX1对滋养细胞的生物学行为的影响,及其参与PE发生的潜在机制。方法:收集2015年11月至2016年12月于重庆医科大学附属第一医院产科住院行选择性剖宫产术的孕妇的胎盘组织,包括正常足月妊娠组(31例)和PE组(35例)。正常氧条件下,人绒毛外滋养细胞系(human transformed primary extravillous trophoblast cell line,HTR8/SVneo)细胞分为小干扰RNA(small interfering RNA,siRNA)阴性对照组(siControl)和QSOX1特异siRNA干扰组(siQSOX1),每组6例样本,实验重复3次。缺氧条件下,HTR8/SVneo细胞分为4组,分别为正常氧处理组(0 h)、缺氧6 h处理组(6 h)、缺氧12 h处理组(12 h)和缺氧24 h处理组(24 h),每组各6例样本,实验重复3次。采用Western blot法检测正常孕妇与PE孕妇胎盘中QSOX1的差异表达以及HTR8/SVneo细胞缺氧处理后QSOX1和缺氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)的蛋白表达。利用siRNA技术干扰HTR8/Svneo细胞QSOX1的表达,用Western blot法检测转染细胞活性半胱氨酸天冬氨酸蛋白酶(Cleaved cysteinyl aspartate specific proteinase,Cleaved caspase)相关蛋白和细胞周期蛋白D1(CyclinD1)的表达水平。并利用流式细胞术检测转染细胞凋亡,比色法检测转染细胞的增殖情况。结果:QSOX1在PE胎盘中的表达明显高于正常胎盘(0.955±0.285 vs. 3.921±0.960)(P=0.001)。缺氧处理HTR8/SVneo细胞后QSOX1和HIF-1α的表达均明显升高(1.183±0.160 vs. 3.987±0.098;1.051±0.444 vs.1.912±0.118)(均P=0.000)。下调QSOX1后HTR8/SVneo细胞Cleaved caspase-3、Cleaved caspase-9蛋白表达明显减少(2.940±0.514 vs. 1.075±0.121;9.223±1.230 vs. 1.011±0.019)(P=0.004;P=0.000),而CyclinD1的蛋白表达增加(1.851±0.972vs. 4.189±0.399)(P=0.018)。QSOX1干扰后HTR8/SVneo细胞凋亡能力减低[(21.007±2.214)%vs.(10.057±0.388)%](P=0.001),增殖能力增加(0.389±0.162
Objective:To explore the expression of quiescin sulfhydryl oxidase 1(QSOX1) in the placentas of preeclampsia(PE),its effects on the apoptosis and proliferation of trophoblast cells and its probable role in the development of PE. Methods:Human term placentas from normal pregnancies(31 cases) and from pregnancies complicated by PE(35 cases) were obtained,and all samples collected in the First Affiliated Hospital of Chongqing Medical University from November 2015 to December 2016. Under the condition of normal oxygen,human transformed primary extravillous trophoblast cell line(HTR8/SVneo) cells were divided into the small interference RNA(siRNA) group,the negative control group(siControl) and the QSOX1 inhibition group(siQSOX1),and the experiment repeated 3 times,6 samples in each group. Under the condition of hypoxia,HTR8/SVneo cells were divided into 4 groups respectively,the normal oxygen treatment group(0 h),the 6 hour hypoxia treatment group(6 h),the 12 hour hypoxia treatment group(12 h) and the 24 hour hypoxia treatment group(24 h),and the experiment repeated 3 times,6 samples in each group. Western blot was used to detect the expression levels of QSOX1 in the placentas from normal pregnant women and from PE pregnant women,and the expression levels of QSOX1 and hypoxia inducible factor-1α(HIF-1α) in human transformed primary extravillous trophoblast(HTR8/SVneo) cells treated with hypoxia. Down-regulation of QSOX1 in HTR-8/Svneo cells was achieved by small interfering RNA(siRNA) interference. The protein expression levels of cleaved cysteinyl aspartate specific proteinases(Cleaved caspase) and CyclinD1 in HTR-8/Svneo cells treated with siRNA were measured by Western blot. Furthermore,the apoptosis of transfected cells was detected by flow cytometry,and the cell proliferation by colorimetric method. Results:QSOX1 was overexpressed in the PE placenta compared with the normal placenta(0.955±0.285 vs. 3.921±0.960) (P=0.001). Hypoxia induced the higher expressions of QSOX1 and HIF-1α in HTR-8/Svneo cells(1.183�
作者
李金津
童超
漆洪波
Li Jinjin;Tong Chao;Qi Hongbo(Department of Obstetrics,The First Affiliated Hospital of Chongqing Medical University,International Collaborative Joint Laboratory of Reproduction and Development of Ministry of Education of China, Chongqing Medical University)
出处
《重庆医科大学学报》
CAS
CSCD
北大核心
2019年第1期6-11,共6页
Journal of Chongqing Medical University
关键词
凋亡
增殖
滋养细胞
子痫前期
静息巯基氧化酶-1
apoptosis
proliferation
trophoblast cells
preeclampsia
quiescin sulfhydryl oxidase 1
