miR-34c works downstream of p53 leading to dairy goat male germline stem-cell (mGSCs) apoptosis
miR-34c works downstream of p53 leading to dairy goat male germline stem-cell (mGSCs) apoptosis
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DOI：
10.1111/cpr.12013
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发表时间：
2013-04-01
影响因子：
8.5
通讯作者：
Hua, J.
中科院分区：
文献类型：
--
作者：
Li, M.;Yu, M.;Hua, J.
目的：最近的证据表明 miR-34c 在细胞周期、细胞衰老和小鼠和人类肿瘤细胞凋亡、精子发生和雄性生殖细胞凋亡的调节中发挥重要作用。然而，关于miR-34c对家畜雄性生殖细胞增殖和凋亡影响的信息很少。奶山羊是一种便于生物学研究和应用的家养品种。本研究的目的是探讨miR-34c对奶山羊雄性种系干细胞（mGSC）凋亡和增殖的影响，并确定该物种中p53与miR-34c之间的关系。材料和方法：形态学观察、miRNA原位杂交（ISH）、溴脱氧尿苷染色、流式细胞术、定量RT-PCR（Q-RT-PCR）和蛋白质印迹法来确定通过转染 miR-34c 模拟物 (miR-34c)、miR-34c 抑制剂 (anti-miR-34c)、miR-34c 模拟物和抑制剂共转染（混合物），与对照组相比，观察 mGSC 的凋亡和增殖。 结果：结果表明 miR-34c 过表达促进 mGSC 凋亡并抑制其增殖。同时，多种凋亡相关基因表达增加，而一些增殖相关基因表达下调。因此，miR-34c 促进 mGSC 凋亡并减少其增殖。此外，miR-34c的表达具有p53依赖性。结论：本研究首次为雄性奶山羊生殖细胞的miRNA及其增殖和凋亡机制的研究提供了模型。
Objectives: Recent lines of evidence have indicated that miR-34c can play important roles in regulation of the cell cycle, cell senescence and apoptosis of mouse and human tumour cells, spermatogenesis, and male germ-cell apoptosis. However, there is little information on the effects of miR-34c on proliferation and apoptosis of livestock male germ cells. The dairy goat is a convenient domestic species for biological investigation and application. The purpose of this study was to investigate the effects of miR-34c on apoptosis and proliferation of dairy goat male germline stem cells (mGSCs), as well as to determine the relationship between p53 and miR-34c in this species.Materials and methods: Morphological observation, miRNA in situ hybridisation (ISH), bromodeoxyuridine staining, flow cytometry, quantitative-RT-PCR (Q-RT-PCR) and western blotting were utilized to ascertain apoptosis and proliferation of mGSCs, through transfection of miR-34c mimics (miR-34c), miR-34c inhibitor (anti-miR-34c), miR-34c mimics and inhibitors co-transfected (mixture) compared to control groups.Results: Results manifested that miR-34c overexpression promoted mGSCs apoptosis and suppressed their proliferation. Simultaneously, a variety of apoptosis-related gene expression was increased while some proliferation-related genes were downregulated. Accordingly, miR-34c promoted apoptosis in mGSCs and reduced their proliferation; moreover, expression of miR-34c was p53-dependent.Conclusions: This study is the first to provide a model for study of miRNAs and mechanisms of proliferation and apoptosis in male dairy goat germ cells.