HMGB3对胎儿生长受限患者胎盘滋养层细胞增殖及凋亡的影响与机制分析

Effect and mechanism of HMGB3 on proliferation and apoptosis of placental trophoblast cells in patients with fetal growth restriction

目的观察HMGB3在胎儿生长受限患者胎盘中的表达,探讨其对滋养层细胞增殖及凋亡的影响与机制分析。方法选取分娩的产妇90例,49例健康产妇为正常组,41例FGR患者为FGR组,采用实时荧光定量PCR检测2组胎盘组织中HMGB3 mRNA的表达。将HMGB3过表达质粒(HMGB3组)或空质粒(对照组)分别转染入人绒毛膜滋养层细胞HTR-8/SVneo中,转染24 h后检测2组细胞的增殖、迁移和凋亡情况。采用String生物信息数据库分析与HMGB3相互作用的蛋白,用实时荧光定量PCR和免疫共沉淀检测2组细胞中预测的结合蛋白mRNA的表达及其与HMGB3的结合情况。结果 HMGB3 mRNA在FGR组胎盘组织中的表达显著低于其在正常组胎盘组织中的表达(P<0.001)。与对照组相比,HMGB3组细胞的增殖活性和细胞迁移率显著增加(P<0.05,P<0.001),细胞凋亡率显著降低(P<0.05)。经String软件预测共有10个蛋白与HMGB3相结合,分别为NDNL2、HMGN2、NUMA1、NUCKS1、GM2A、H2AFZ、SUPT16H、SSRP1、BTAF1和NPL。在HMGB3组细胞中,GM2A和NPL表达无变化,NDNL2下调,其余蛋白均上调。结论 HMGB3在FGR患者胎盘组织中低表达,且HMGB3过表达可促进滋养层细胞增殖及迁移,抑制凋亡。HMGB3介导的DNA、组蛋白、染色质等活动异常可能是导致FGR发病的机制之一。

Objective To observe the expression of high mobility group box 3 (HMGB3) in the placentas of patients with fetal growth restriction (FGR),and to explore its effect on proliferation and apoptosis of trophoblast cells and analyze its mechanism.Methods A total of 90 parturients were selected,with 49 healthy ones as the normal group,and 41 FGR patients as the FGR group.The expression of HMGB3 mRNA in the placenta tissues of the two groups was detected by the real-time fluorescent quantitative PCR technology.HMGB3 overexpressed plasmid (HMGB3 group) or empty plasmid (control group) was transfected into human choriotrophoblast cells HTR-8/SVneo,respectively.24 hours later after transfection,the proliferation,migration and apoptosis of the two groups were detected.The proteins interacting with HMGB3 were analyzed with the String bioinformatics database,and the mRNA expression of the predicted binding proteins and their binding to HMGB3 in the cells of the two groups were detected by real-time quantitative PCR and immunoprecipitation.Results The expression of HMGB3 mRNA in the placental tissues of the FGR group was significantly lower than that in the normal group (P<0.001).Compared with the control group,the proliferation activity (P<0.05) and cell migration rate (P<0.001) of the HMGB3 group were significantly increased,and the apoptosis rate (P<0.05) was significantly decreased.With the aid of String software,a total of 10 proteins were predicted to be combined with HMGB3,including NDNL2,HMGN2,NUMA1,NUCKS1,GM2A,H2AFZ,SUPT16H,SSRP1,BTAF1 and NPL.Moreover,in the HMGB3 group,GM2A and NPL expressions did not undergo any changes,and NDNL2 was down-regulated while the remaining proteins were up-regulated.Conclusion HMGB3 is underexpressed in placental tissues of the FGR patients,and overexpression of HMGB3 promotes proliferation and migration of trophoblast cells and inhibits apoptosis.The abnormal activities of DNA,histones,and chromatin mediated by HMGB3 may be one of the mechanisms leading to the pathogenesis of FGR.