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河北医学  2023, Vol. 29 Issue (9): 1432-1437    DOI: 10.3969/j.issn.1006-6233.2023.09.05
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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
黄勇, 罗书, 廖源, 王开举
海南医学院第二附属医院, 海南 海口 570311
HIF-1 Regulates PAD4-Mediated H3cit and Promotes Apoptosis of Endometrial Glandular Epithelial Cells under Hypoxia
HUANG Yong, LUO Shu, LIAO Yuan, et al
The Second Affiliated Hospital of Hainan Medical College, Hainan Haikou 570311, China
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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。
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黄勇
罗书
廖源
王开举
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    
AbstractObjective: 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.
Key wordsAnoxic environment    Anoxic-inducing factor 1    Arginine deiminase 4    Endometrial gland epithelial cells    Apoptosis
    
基金资助:海南省卫生健康行业科研项目,(编号:22A200095)
引用本文:   
黄勇, 罗书, 廖源, 王开举. 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[J]. 河北医学, 2023, 29(9): 1432-1437.
HUANG Yong, LUO Shu, LIAO Yuan, et al. HIF-1 Regulates PAD4-Mediated H3cit and Promotes Apoptosis of Endometrial Glandular Epithelial Cells under Hypoxia. HeBei Med, 2023, 29(9): 1432-1437.
链接本文:  
http://www.hbyxzzs.cn/CN/10.3969/j.issn.1006-6233.2023.09.05     或     http://www.hbyxzzs.cn/CN/Y2023/V29/I9/1432
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