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2009, Volume 3, Issue 4

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Frontiers of Medicine >> 2009, Volume 3, Issue 4 doi: 10.1007/s11684-009-0073-y

Effect of PRAK gene knockout on the proliferation of mouse embryonic fibroblasts

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;

Available online: 2009-12-05
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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 and 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 cells; moreover, the number of cells was decreased by 44.5% compared with that of cells cultured for 96h, suggesting that G/M transition is inhibited in cells. Meanwhile, G/S transition was also inhibited in cells, observed with flow cytometry analysis. The ratios of G/G, G/M, and S phases of cells were 44.9%, 12.2%, and 42.9%, respectively, while those of cells were 55.3%, 7.3%, and 37.4%, respectively. There were 23.1% increase and 12.7% decrease of the number of cells in G and S phases comparison with that of cells, respectively. Taken together, gene knockout in MEF cells leads to cell cycle arrest and proliferation inhibition.

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