STAT3介导甲基化抑制剂5-AZA-DC调控子痫前期滋养细胞凋亡、增殖、侵袭与迁移的作用及分析  

The role and analysis of STAT3 mediatedregulation of trophoblast apoptosis,proliferation,invasion,and migration by methylation inhibitor 5-AZA-DC in preeclampsia

作　　者：黄少娥 谢丽清 丁枝珠 赵晓勇 张晓丽 HUANG Shao′e;XIE Liqing;DING Zhizhu;ZHAO Xiaoyong;ZHANG Xiaoli(Department of Reproductive Medicine,Maternal and Child Health Hospital,Huadu District,Guangzhou 510800,Guangdong,China;不详)

机构地区：[1]广州市花都区妇幼保健院生殖医学科,510800 [2]暨南大学医学部-广东省第二人民医院妇产科 [3]广州医科大学附属第五医院外科

出　　处：《中国生育健康杂志》2024年第6期521-530,共10页Chinese Journal of Reproductive Health

基　　金：广东省自然科学基金-粤企面上(2021A1515220097,2023A1515220108);广州市基础-应用基础科技项目(202201010829,2023A03J0822);广东省哲学社科项目(GD24CJY30)。

摘　　要：目的探讨甲基转移酶抑制剂5-氮杂-2-脱氧胞苷(5-AZA-DC)对子痫前期滋养细胞凋亡、增殖、侵袭与迁移的调控。方法收集40例正常妊娠组产妇和40例子痫前期(PE)组产妇胎盘组织,采用分娩后胎盘HE染色,胎盘组织中全基因组甲基化测序(WGBS),体外培养HTR8/Svneo、JEG-3、JAR、BeWo滋养细胞株,分为空白对照组(Control组)、STAT3沉默组(shSTAT3组)、STAT3过表达组(STAT3^(OE)组)。应用5-AZA-DC处理,采用CCK8、流式细胞技术、Transwell侵袭和细胞划痕实验,检测滋养细胞凋亡、增殖、侵袭与迁移。采用t检验和单因素方差分析,对数据进行统计分析。结果(1)在胎盘组织中,与正常妊娠组相比,PE组血管腔变小,血管闭塞,血流量少,绒毛间质间纤维蛋白沉积,绒毛可见梗死、玻璃样变。WGBS检测出PE组甲基化水平显著增高,其中STAT3[(17.25±1.05)vs.(30.36±1.33),P<0.01]、PTEN[(13.33±0.71)vs.(23.05±1.17),P<0.02]、TSC2[(15.28±1.43)vs.(26.53±1.15),P<0.02]基因为甲基化差异显著基因,差异有统计学意义。(2)在细胞株中,与空白对照组相比,5-AZA-DC干预后滋养细胞凋亡减少、增殖上调、侵袭与迁移上调,差异有统计学意义。结论STAT3基因是引起子痫前期发病的重要基因,5-AZA-DC干预对子痫前期滋养细胞凋亡、增殖、侵袭与迁移作用和影响显著。5-AZA-DC可能成为介导表观遗传学治疗子痫前期的潜在药物。Objective To investigate the regulation of the methyltransferase inhibitor 5-aza-2-deoxycytidine(5-AZA-DC)on trophoblast apoptosis,proliferation,invasion,and migration in preeclampsia.Methods Placental tissue samples were collected from 40 normal pregnancies(normal pregnancy group)and 40 preeclampsia(PE group)patients.Hematoxylin-eosin(HE)staining was performed on the placental tissues after delivery,and whole-genome bisulfite sequencing(WGBS)was conducted on the placental tissues.Trophoblast cell lines HTR8/Svneo,JEG-3,JAR,and BeWo were cultured in vitro and divided into blank control group(Control group),STAT3 silencing group(shSTAT3 group),and STAT3 overexpression group(STAT3^(OE) group).Cells were treated with 5-AZA-DC,and cell apoptosis,proliferation,invasion,and migration were assessed using CCK8,flow cytometry,Transwell invasion assays,and scratch assays.Statistical analysis was performed using t-tests and one-way ANOVA.Results(1)In placental tissue,compared to the normal pregnancy group,the PE group showed reduced vascular lumens,vascular occlusion,blood flow,fibrin deposition in the intervillous stroma,and visible infarction and hyaline degeneration in the villi.WGBS revealed a significant increase in methylation levels in the PE group,with statistically significant differences in the methylation of genes such as STAT3[(17.25±1.05 vs 30.36±1.33),P<0.01],PTEN[(13.33±0.71 vs 23.05±1.17),P<0.02],TSC2[(15.28±1.43 vs 26.53±1.15),P<0.02].(2)In the cell lines,compared to the Control group,5-AZA-DC intervention significantly reduced trophoblast apoptosis while increasing cell proliferation,invasion,and migration.These differences were statistically significant(P<0.05).Conclusion STAT3 is a key gene in the pathogenesis of preeclampsia.The intervention with 5-AZA-DC intervention has significant effects on trophoblast apoptosis,proliferation,invasion,and migration in preeclampsia.5-AZA-DC may be a potential epigenetic therapeutic agent for preeclampsia.

关 键 词：5-氮杂-2-脱氧胞苷(5-AZA-DC) 滋养细胞 凋亡和增殖 侵袭与迁移 调控 

分 类 号：R714.24[医药卫生—妇产科学]