人真核翻译起始因子4A3调控的环状RanGTP酶激活蛋白1通过调节滋养细胞的增殖、迁移和侵袭参与子痫前期发病  

Eukaryotic translation initiation factor 4A3-regulated circular RanGTPase activating protein 1 participates in pathogenesis of preeclampsia by regulating proliferation,migration,and invasion of trophoblast cells

作　　者：王婷婷[1,2,3,4] 葛云鹏 沈鸿飞[1,2,3,4] 李佳钋 刘奕霖 乔宠 Wang Tingting;Ge Yunpeng;Shen Hongfei;Li Jiapo;Liu Yilin;Qiao Chong(Department of Obstetrics and Gynecology,Shengjing Hospital of China Medical University,Shenyang 110004,China;Key Laboratory of Reproductive and Genetic Medicine,National Health Commission,Shenyang 110004,China;Key Laboratory of Maternal-Fetal Medicine of Liaoning Province,Benxi 117000,China;Research Center of China Medical University Birth Cohort,Shenyang 110004,China)

机构地区：[1]中国医科大学附属盛京医院妇产科,沈阳110004 [2]国家卫生健康委员会生殖健康与遗传医学重点实验室,沈阳110004 [3]辽宁省母胎医学重点实验室,本溪117000 [4]中国医科大学出生队列研究中心,沈阳110004

出　　处：《中华围产医学杂志》2024年第9期742-749,共8页Chinese Journal of Perinatal Medicine

基　　金：国家重点研发计划(2016YFC1000404);国家自然基金面上项目(81370735);辽宁省民生科技计划联合计划项目(2021JH2/10300123);沈阳市科技计划项目(20-205-4-004)。

摘　　要：目的探讨环状RanGTP酶激活蛋白1(circular RanGTPase activating protein 1,circRANGAP1)在子痫前期中对滋养细胞生物学行为的影响及其可能的作用机制。方法收集2020年8月至2022年12月于中国医科大学附属盛京医院就诊的子痫前期患者及年龄、孕周匹配的对照组产妇的胎盘组织(早发子痫前期组和早发对照组各8例,晚发子痫前期和晚发对照组各24例),采用实时荧光定量法检测胎盘组织中circRANGAP1及人真核翻译起始因子4A3(eukaryotic translation initiation factor 4A3,EIF4A3)mRNA表达水平,蛋白质印迹法检测EIF4A3蛋白表达水平。取滋养细胞系HTR-8/Svneo,采用细胞计数增殖试验、划痕试验和Transwell侵袭试验检测增殖、迁移和侵袭能力,采用RNA结合蛋白免疫沉淀法检测EIF4A3对circRANGAP1的调控作用。敲降EIF4A3表达后,实时荧光定量聚合酶链反应法检测HTR-8/Svneo细胞中circRANGAP1的变化。采用独立样本t检验、非参数χ^(2)检验或Pearson相关分析对数据进行统计学分析。结果(1)早发子痫前期中circRANGAP1表达与早发对照组差异无统计学意义,晚发子痫前期中circRANGAP1表达高于晚发对照组(3.764±3.297与0.960±0.720,t=4.07,P<0.001)。在晚发子痫前期中,circRANGAP1的表达与收缩压及舒张压均呈正相关(收缩压:r=0.639,P<0.01;舒张压:r=0.800,P<0.001)。早发子痫前期中EIF4A3 mRNA及蛋白质表达与早发对照组差异无统计学意义,晚发子痫前期中EIF4A3 mRNA及蛋白质表达高于晚发对照组(mRNA:2.963±3.081与1.149±0.667,t=2.30,P=0.028;蛋白质:2.504±1.008与0.258±0.180,t=4.39,P=0.005)。(2)小干扰(small interfering,siRNA)敲降后,mRANGAP1的表达量差异无统计学意义,而circRANGAP1的表达量降低(1.000±0.004、0.465±0.031与0.621±0.030),其中si-1敲降效率最高(t=23.59,P=0.002)。特异性敲降circRANGAP1后,滋养细胞的增殖(1.297±0.058与1.456±0.030,t=-5.97,P<0.001)、侵袭(94.400±6.504与219.000±19.870,t=-13.32,P<ObjectiveTo investigate the impact of circular RanGTPase activating protein 1(circRANGAP1)on the biological behavior of trophoblast cells in preeclampsia and its potential mechanisms.MethodsPlacental tissues were collected from preeclampsia patients and age-and gestational age-matched control pregnant women admitted to Shengjing Hospital of China Medical University from August 2020 to December 2022(eight cases each in the early-onset preeclampsia group and early-onset control group,and 24 cases each in the late-onset preeclampsia group and late-onset control group).The expression levels of circRANGAP1 and eukaryotic translation initiation factor 4A3(EIF4A3)mRNA in placental tissues were detected by real-time quantitative polymerase chain reaction(RT-qPCR),and EIF4A3 protein expression was assessed by Western blotting.In HTR-8/Svneo cells,the proliferation,migration,and invasion abilities were evaluated by cell counting assay,scratch assay,Transwell invasion assay,and the regulatory effect of EIF4A3 on circRANGAP1 was examined by RNA binding protein immunoprecipitation(RIP).Changes of circRANGAP1 expression in HTR-8/Svneo cells were detected by RT-qPCR after EIF4A3 knockdown.Statistical analysis was performed using independent sample t-test,non-parametric Chi-square test,or Pearson correlation analysis.Results(1)There was no significant difference in circRANGAP1 expression between the early-onset preeclampsia group and the early-onset control group.However,circRANGAP1 expression was higher in the late-onset preeclampsia group compared to the late-onset control group[(3.764±3.297)vs.(0.960±0.720),t=4.07,P<0.001].In late-onset preeclampsia patients,circRANGAP1 expression was positively correlated with both systolic and diastolic blood pressure(systolic:r=0.639,P<0.01;diastolic:r=0.800,P<0.001).There was no significant difference in EIF4A3 mRNA and protein expression between the early-onset preeclampsia group and the early-onset control group,but EIF4A3 mRNA and protein expression were higher in the late-onset pree

关 键 词：子痫前期 circRANGAP1 滋养细胞 EIF4A3 

分 类 号：R714.244[医药卫生—妇产科学]