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 manifested that miR-34c over-expression 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.
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.