M Li1, M Yu, C Liu, H Zhu, X He, S Peng, J Hua. 1. College of Veterinary Medicine, Shaanxi Centre of Stem Cells Engineering & Technology, Key Lab for Animal Biotechnology of Agriculture Ministry, Northwest A&F University, Yangling, Shaanxi, 712100, China.
Abstract
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 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. 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.
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 humantumour 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 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. 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.
Authors: Lin Zhang; Jia Huang; Nuo Yang; Joel Greshock; Molly S Megraw; Antonis Giannakakis; Shun Liang; Tara L Naylor; Andrea Barchetti; Michelle R Ward; George Yao; Angelica Medina; Ann O'brien-Jenkins; Dionyssios Katsaros; Artemis Hatzigeorgiou; Phyllis A Gimotty; Barbara L Weber; George Coukos Journal: Proc Natl Acad Sci U S A Date: 2006-06-05 Impact factor: 11.205
Authors: David C Corney; Andrea Flesken-Nikitin; Andrew K Godwin; Wei Wang; Alexander Yu Nikitin Journal: Cancer Res Date: 2007-09-06 Impact factor: 12.701