缺氧环境下HIF-1调控PAD4介导的H3cit促进子宫内膜腺上皮细胞凋亡  

HIF-1 Regulates PAD4-Mediated H3cit and Promotes Apoptosis of Endometrial Glandular Epithelial Cells under Hypoxia

作　　者：黄勇[1] 罗书[1] 廖源 王开举 HUANG Yong;LUO Shu;LIAO Yuan(The Second Affiliated Hospital of Hainan Medical College,Hainan Haikou 570311,China)

机构地区：[1]海南医学院第二附属医院,海南海口570311

出　　处：《河北医学》2023年第9期1432-1438,共7页Hebei Medicine

基　　金：海南省卫生健康行业科研项目,(编号:22A200095)。

摘　　要：目的:探究缺氧环境下HIF-1以及PAD4对子宫内膜腺上皮细胞凋亡的作用及其相关机制。方法:常氧和缺氧环境培养宫内膜腺上皮细胞CM-H058,qRT-PCR和Western blot检测HIF-1、PAD4、H3cit、Caspase3、Caspase7表达,流式细胞术检测细胞凋亡水平。通过慢病毒转染法将sh-NC、sh-HIF-1、oe-NC、oe-PAD4转染至CM-H058,缺氧环境培养并分组为:sh-NC组、sh-HIF-1组、sh-HIF-1+oe-NC组和sh-HIF-1+oe-PAD4组,检测各组细胞HIF-1、PAD4、H3cit、Caspase3、Caspase7表达和细胞凋亡水平;检测PAD4、H3cit和H3K27ac在Caspase3、Caspase7 TSS富集水平。结果:与常氧组比较,缺氧组HIF-1、PAD4、H3cit、Caspase3、Caspase7表达升高(P<0.05),细胞凋亡水平升高(P<0.05)。与sh-NC组比较,sh-HIF-1组HIF-1、PAD4、H3cit、Caspase3、Caspase7表达降低(P<0.05),细胞凋亡水平降低(P<0.05),HIF-1在PAD4 TSS富集水平降低(P<0.05)。与sh-HIF-1+oe-NC组比较,sh-HIF-1+oe-PAD4组PAD4、H3cit、Caspase3、Caspase7表达升高(P<0.05),细胞凋亡水平升高(P<0.05)。结论:缺氧环境下HIF-1促进PAD4表达。PAD4及其催化的H3cit进一步在Caspase3、Caspase7 TSS富集,促进Caspase3、Caspase7转录和表达,最终诱导子宫内膜腺上皮细胞凋亡。Objective:To investigate the effects of HIF-1 and PAD4 on apoptosis of endometrial epithelial cells under hypoxia and its related mechanisms.Methods:Endometrial glandular epithelial cells CM-H058 were cultured under normal and anoxic conditions,and the expressions of HIF-1,PAD4,H3cit,Caspase3 and Caspase7 were detected by qRT-PCR and Western blot.The apoptosis level of cells was detected by flow cytometry.sh-NC,sh-HIF-1,oe-NC and oe-PAD4 were transfected into CM-H058 by lentivirus transfection method,then cultured in anoxic environment and divided into sh-NC group,sh-HIF-1 group,sh-HIF-1+oe-NC group,and sh-HIF-1+oe-PAD4 group.The expressions of HIF-1,PAD4,H3cit,Caspase3,Caspase7 and apoptosis of cells in each group were detected.The concentration of PAD4,H3cit,and H3K27ac in TSS of Caspase3 and Caspase7 were detected.Results:Compared with the normal oxygen group,the expressions of HIF-1,PAD4,H3cit,Caspase3 and Caspase7 in the hypoxia group were increased(P<0.05),and the apoptosis level was increased(P<0.05).Compared with the sh-NC group,the expressions of HIF-1,PAD4,H3cit,Caspase3 and Caspase7 in sh-HIF-1 group were decreased(P<0.05),and the apoptosis level was decreased(P<0.05).Compared with sh-HIF-1+oe-NC group,the expressions of PAD4,H3cit,Caspase3,and Caspase7 in sh-HIF-1+oe-PAD4 group were increased(P<0.05),and the apoptosis level was increased(P<0.05),and the enrichment level of PAD4,H3cit and H3K27ac in TSS of Caspase3,and Caspase7 were increased(P<0.05).Conclusion:HIF-1 promotes PAD4 expression.PAD4 and its catalyzed H3cit further enriched Caspase3 and Caspase7 TSS,promoted the transcription and expression of Caspase3 and Caspase7,and finally induced apoptosis of endometrial epithelial cells.

关 键 词：缺氧环境 缺氧诱导因子1 精氨酸脱亚胺酶4 子宫内膜腺上皮细胞 凋亡 

分 类 号：R711[医药卫生—妇产科学]