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Effect of PRAK gene knockout on the proliferation of mouse embryonic fibroblasts |
Xiaowei GONG MD, PhD1,Xiaoyan MING MD1,Xu WANG MM1,Daan WANG MD1,Peng DENG MM1,Yong JIANG MD, PhD1,Aihua LIU MD, PhD2, |
1.Department of Pathophysiology,
Key Laboratory of Proteomics of Guangdong Province, Southern Medical
University, Guangzhou 510515, China; 2.Department of Respiratory
Diseases, Nanfang Hospital, Southern Medical University, Guangzhou
510515, China; |
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Abstract p38 regulated/activated protein kinase (PRAK) plays a key role in cell senescence and tumor suppression. The aim of this study was to investigate if PRAK had effect on cell proliferation. The growth of PRAK+/+ and PRAK−/− mouse embryonic fibroblast (MEF) cells was measured by methylthiazoletetrazolium (MTT) colorimetric assay, and the proportion of the cell number in different phases of the cell cycle was analyzed by flow cytometry. The growth curves showed that the growth rate was notably decreased, and cell double time was elongated in PRAK−/− cells; moreover, the number of PRAK−/− cells was decreased by 44.5% compared with that of PRAK+/+ cells cultured for 96h, suggesting that G2/M transition is inhibited in PRAK−/− cells. Meanwhile, G1/S transition was also inhibited in PRAK−/− cells, observed with flow cytometry analysis. The ratios of G0/G1, G2/M, and S phases of PRAK+/+ cells were 44.9%, 12.2%, and 42.9%, respectively, while those of PRAK−/− cells were 55.3%, 7.3%, and 37.4%, respectively. There were 23.1% increase and 12.7% decrease of the number of PRAK−/− cells in G1 and S phases comparison with that of PRAK+/+ cells, respectively. Taken together, PRAK gene knockout in MEF cells leads to cell cycle arrest and proliferation inhibition.
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Keywords
p38 regulated/activated protein kinase
gene knockout
cell cycle
cell proliferation
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Issue Date: 05 December 2009
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