miR-3074-5p Promotes the Apoptosis but Inhibits the Invasiveness of Human Extravillous Trophoblast-Derived HTR8/SVneo Cells In Vitro
miR-3074-5p Promotes the Apoptosis but Inhibits the Invasiveness of Human Extravillous Trophoblast-Derived HTR8/SVneo Cells In Vitro
复制标题
miR-3074-5p 在体外促进人绒毛外滋养层来源的 HTR8/SVneo 细胞凋亡但抑制侵袭性
DOI：
10.1177/1933719117725823
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发表时间：
2018-05-01
影响因子：
2.9
通讯作者：
Wang, Jian
中科院分区：
文献类型：
--
作者：
Gu, Yan;Shi, Yan;Wang, Jian
目的：观察miR-3074-5p过表达对体外培养的人滋养层细胞的影响。设计：HTR8/SVneo细胞体外实验研究。流式细胞仪检测细胞凋亡率，Transwell法检测细胞侵袭力。用Western印迹法检测HTR8/SVneo细胞中P53、细胞周期蛋白依赖性激酶抑制因子1B(P27)、bcl2、bcl2相关基因X(Bax)和bcl2样蛋白14(BclG)的表达水平。采用基因芯片技术检测HTR8/SVneo细胞基因表达谱变化，Western印迹法检测二氢硫胺S琥珀酰基转移酶(DLST)、生长相关蛋白43(GAP43)、矮小相关转录因子2(RUNX2)和C-C型趋化因子受体3(CCR3)的差异表达。结果：在HTR8/SVneo细胞中，miR-3074-5p的过表达促进了细胞的凋亡，但抑制了细胞的侵袭，并伴随着P27、bcl2和bclG的表达显著升高。同时，复发性流产(RM)患者胎盘绒毛中P27和P57的表达也增加。总共筛选出411个基因和397个基因在高表达HTR8/SVneo细胞中至少上调2倍或下调2倍。这些差异表达的基因参与了与怀孕相关的几个重要功能。随后，在过表达的HTR8/SVneo细胞中，DLST和GAP43蛋白的表达降低，CCR3和RUNX2蛋白的表达增加。结论：miR-3074-5p可能通过干扰滋养层细胞的正常活动而在RM的发病机制中发挥作用。
Objective: The objective of this study was to observe the effects of the overexpression of miR-3074-5p in human trophoblast cells in vitro.Design: Experimental in vitro study in HTR8/SVneo cells.Methods: HTR8/SVneo cells were transfected with miR-3074-5p mimic. The cell apoptosis and invasion were measured via flow cytometry and transwell assay, respectively. The expression levels of P53, Cyclin Dependent Kinase Inhibitor 1B (P27), BCL-2, BCL2 associated X (BAX), and BCL2 like 14 (BCL-G) in HTR8/SVneo cells were determined by Western blot. The alterations in gene expression profile of HTR8/SVneo cells were evaluated by complementary DNA microarray assay, and the differential expressions of dihydrolipoamide S-succinyltransferase (DLST), growth-associated protein 43 (GAP43), runt-related transcription factor 2 (RUNX2), and C-C type chemokine receptor 3 (CCR3) were validated by Western blot. Biofunctions of these differentially expressed genes were enriched by Gene Ontology analysis.Results: The overexpression of miR-3074-5p in HTR8/SVneo cells promoted cell apoptosis but inhibited cell invasion, being accompanied by the significantly elevated expressions of P27, BCL-2, and BCL-G. Meanwhile, an increased expression of P27 and P57 was also detected in a small sample size of placental villi of recurrent miscarriage (RM) patients. Totally, 411 genes and 397 genes were screened out, respectively, to be downregulated or upregulated at least by 2-folds in miR-3074-5p overexpressed HTR8/SVneo cells. These differentially expressed genes were involved in several important functions related to pregnancy. Subsequently, the reduced expressions of DLST and GAP43 proteins, as well as the increased expressions of CCR3 and RUNX2 proteins, were validated in miR-3074-5p overexpressed HTR8/SVneo cells.Conclusion: These data suggested a potential contribution of miR-3074-5p in the pathogenesis of RM by disturbing the normal activities of trophoblast cells.