Silencing caspase-3 gene effects on the proliferation and apoptosis of rat bone marrow mesenchymal stem cells

doi:10.3969/j.issn.2095-4344.2013.14.002

Abstract

Abstract:

BACKGROUND: Ischemia and hypoxia myocardial microenvironment leads to poor survival of transplanted cells.
OBJECTIVE: To investigate the effects of silencing caspase-3 gene on proliferation and apoptosis of rat bone marrow mesenchymal stem cells under ischemia and hypoxia in vitro.
METHODS: Lentiviral short hairpin RNA interference vector targeting caspase-3 was constructed and transfected into mesenchymal stem cells as the transgene group. The normal cell group and the empty vector group were as controls. MTS assay was applied to examine the proliferation of cells in each group. The ischemia and hypoxia model was established. The expressions of caspase-3 mRNA and protein were detected by real-time PCR and immunohistochemistry. The apoptotic rates of the cells at different time points (0, 6, 12, 24 and 48 hours) in each group were evaluated by flow cytometry.
RESULTS AND CONCLUSION: Recombinant lentivirus was transfected into mesenchymal stem cells successfully and the proliferation activity of the cells was increased (P < 0.05). Compared with control groups, the levels of caspase-3 mRNA and protein in the transgene group were decreased (P < 0.05) under ischemia and hypoxia. Silencing caspase-3 could reduce the apoptotic rate of mesenchymal stem cells (P < 0.05), and the apoptotic rate was increased slowly as ischemia and hypoxia time prolonging. Silencing caspase-3 can increase the growth speed and the anti-apoptosis ability of mesenchymal stem cells under ischemia and hypoxia in vitro.

Key words: stem cells, bone marrow-derived stem cells, bone marrow mesenchymal stem cells, gene silencing, Caspase-3, gene, cell proliferation, apoptosis, lentivirus, ischemia and hypoxia, short hairpin RNA, provincial grants-supported paper, stem cell photographs-containing paper

CLC Number:

R394.2

Liu Jia-liang, Hua Ping, Yang Song-ran, Tao Jun, Jiang Hui-qi, Wang Meng, Yang Yan-qi. Silencing caspase-3 gene effects on the proliferation and apoptosis of rat bone marrow mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(14): 2480-2487.

Figures/Tables 7

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