摘要
目的:探讨肿瘤蛋白P53(TP53)在不明原因复发性流产(URSA)患者绒毛组织中的表达及其对人滋养层细胞凋亡和迁移的影响,初步阐明其机制。方法:选取40例URSA患者(患者组)和30例实施人工流产健康孕妇(对照组)的绒毛组织,采用RT-PCR和Western blotting法检测绒毛组织中TP53mRNA及蛋白表达水平。培养人滋养层HTR-8细胞,分为空白对照组、空载体组和TP53-siRNA组,Western blotting法检测转染siRNA后3组细胞中TP53蛋白表达水平。流式细胞术、Transwell小室和Western blotting法分别检测空白对照组和TP53-siRNA组细胞凋亡率、细胞迁移数和细胞中TP53、Cleaved-Caspase3、Bcl-2和Bax蛋白表达水平。结果:患者组绒毛组织中TP53mRNA和蛋白表达水平均明显高于对照组(P<0.01)。TP53-siRNA组细胞中TP53蛋白表达水平明显低于空白对照组(P<0.01)。细胞转染48 h后,与空白对照组比较,TP53-siRNA组细胞凋亡率明显降低(P<0.01),细胞迁移数明显升高(P<0.01),Cleaved-Caspase3和Bax蛋白表达水平均明显降低(P<0.01),Bcl-2蛋白表达水平明显升高(P<0.01)。结论:TP53在URSA患者绒毛组织中呈高表达,沉默人滋养层HTR-8细胞TP53表达能明显促进细胞迁移和抑制细胞凋亡。
Objective：To investigate the expression of tumor protein P53（TP53）in the villus tissue of the patients with unexplained recurrent spontaneous abortion（URSA）and the effects on the apoptosis and migration of human trophoblast cells,and to clamify their mechanisms preliminarily.Methods：The villus tissues of 40 patients with URSA（patient group）and 30 healthy pregnant women underwent artificial abortion（control group）were selected.The expression levels of TP53 mRNA and protein in the villus tissue were detected by RT-PCR and Western blotting method.The human trophoblast HTR-8 cells were divided into blank control group,vector group and TP53-siRNA group.Western blotting method was employed to detect the expression levels of TP53 protein in cells in three groups after transfected with siRNA.The apoptotic rates,the number of migrated cells and the expression levels of TP53,Cleaved-Caspase3,Bcl-2,and Bax proteins in blank control group and TP53-siRNA group were detected respectively by flow cytometry,Transwell assay and Western blotting method.Results：The mRNA and protein expression levels of TP53 in the villus tissue of the patients in patient group were significantly higher than those in control group（P〈0.01）.The TP53 protein expression level in cells in TP53-siRNA group was significantly lower than that in blank control group（P〈0.01）.After transfection for 48 h,the apoptotic rate in TP53-siRNA group was significantly lower than that in blank control group,the number of migrated cells was significantly higher than that in blank control group（P〈0.01）,the expression levels of Cleaved-Caspase3 and Bax proteins in TP53-siRNA group were significantly lower than that in blank control group,and the expression level of Bcl-2 protein was significantly higher than that in blank control group（P〈0.01）.Conclusion：TP53 highly expresses in the villus tissue of the patients with URSA;silencing of TP53 expression in HTR-8 cells can significantly promote the migration of human trophoblast c
作者
朱效慧
孟琴
冯世燕
ZHU Xiaohui;MENG Qin;FENG Shiyan(Department of Obstetrics and Gynecology,People＇s Hospital of Linyi,Shandong Province,Linyi 276000,China;Department of Obstetrics and Gynecology,Linyi Medical College,Linyi 276000,China)
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2018年第4期796-800,共5页
Journal of Jilin University：Medicine Edition
基金
山东省卫计委卫生科研规划项目资助课题(2013WS1701)
关键词
肿瘤蛋白P53
不明原因复发性流产
细胞增殖
细胞凋亡
细胞迁移
tumor protein P53
unexplained recurrent spontaneous abortion
cell proliferation
apoptosis
cell migration
