摘要
目的:探讨低氧对滋养细胞HIF-1α、BNIP3表达及自噬和侵袭能力的影响。方法:体外培养滋养细胞HTR-8/SVneo,分为常氧组、缺氧组和3-MA(自噬抑制剂)+缺氧组。24 h后,分别采用Real-time PCR和Western blot检测常氧组与缺氧组滋养细胞HIF-1α、BNIP3 mRNA和蛋白的表达水平;采用Western blot检测3组滋养细胞自噬标志蛋白Beclin 1和LC3蛋白的表达水平,并用Transwell侵袭实验测定细胞的侵袭能力。结果:与常氧组相比,缺氧组BNIP3 mRNA及HIF-1α、BNIP3蛋白的表达水平升高(P<0.05),但HIF-1αmRNA的表达水平无明显变化(P>0.05)。与常氧组相比,缺氧组Beclin 1、LC3Ⅱ/Ⅰ蛋白的表达水平升高,侵袭细胞数增加;与缺氧组相比,3-MA+缺氧组Beclin 1和LC3Ⅱ/Ⅰ蛋白表达水平降低,侵袭细胞数减少(P<0.05)。结论:低氧环境可能通过HIF-1α/BNIP3途径诱导滋养细胞自噬水平增强,并可以增强其侵袭能力;低氧环境下抑制自噬后滋养细胞侵袭性减弱。
Aim：To investigate the effects of hypoxia on the expressions of HIF-1α and BNIP3,and assess the autophagy and invasion ability of trophoblasts.Methods：First,HTR-8/SVneo cells were divided into control group,hypoxia group,and autophagy inhibitor 3-MA plus hypoxia group（3-MA＋ hypoxia group）.After 24 h,Real-time PCR and Western blot were utilized to measure the expression levels of HIF-1α and BNIP3 mRNA and protein in the first 2 groups.Then the expression levels of Beclin1 and LC3 protein as well as the invasion activity of the cells were detected by Western blot and Transwell Matrigel assay.Results：Compared with control group,the expression level of HIF-1α mRNA in hypoxia group had no significant difference（P〉0.05）;however,the expression levels of BNIP3 mRNA and protein and that of HIF-1α protein in hypoxia group were increased apparently（P〈0.05）.Compared with control group,the expression levels of Beclin 1 and LC3Ⅱ/Ⅰprotein and the invasion ability of trophoblasts increased（P〈0.05）;compared with hypoxia group,the expression levels of Beclin 1 and LC3Ⅱ/Ⅰ protein decreased,and the invasion ability was reduced in 3-MA＋hypoxia group（P〈0.05）.Conclusion：Hypoxia could induce autophagy and enhance the invasion ability of trophoblasts via the HIF-1α/BNIP3 pathway;the invasion ability of trophoblasts under hypoxia is decreased in the presence of 3-MA.
作者
闫广伟
丁燕子
邢金芳
代延朋
杜红梅
袁恩武
YAN Guangwei;DING Yanzi;XING Jinfang;DAI Yanpeng;DU Hongmei;YUAN Enwu(Department of Clinical Laboratory, the Third Affiliated Hospital, Zhengzhou University, Zhengzhou 45005)
出处
《郑州大学学报（医学版）》
CAS
北大核心
2018年第2期198-202,共5页
Journal of Zhengzhou University(Medical Sciences)
基金
河南省科技厅创新项目(20140269)
