p38丝裂原活化蛋白激酶基因敲除对小鼠胚胎成纤维细胞增殖的影响  

Effect of p38 mitogen-activated protein kinase gene knockout on cell proliferation of embryonic fibroblasts in mice

作　　者：龚小卫[1] 魏洁[1] 李煜生[1] 程蔚蔚[1] 邓鹏[1] 姜勇[1] 

机构地区：[1]南方医科大学病理生理学教研室,广东省蛋白质组学重点实验室,广东广州510515

出　　处：《中国危重病急救医学》2008年第9期527-529,共3页Chinese Critical Care Medicine

基　　金：国家自然科学基金资助项目（30670828,30572151）;广东省科技计划项目（A1090202）

摘　　要：目的研究p38丝裂原活化蛋白激酶（MAPK）基因敲除对小鼠胚胎成纤维细胞增殖的影响。方法用蛋白质免疫印迹法（Western blotting）检测小鼠胚胎成纤维细胞中p38制^＋/＋和p380^-/-的表达；利用噻唑蓝（MTT）比色法绘制p38^＋/＋和p38^-/-细胞的生长曲线，用流式细胞仪检测细胞周期各时相所占百分比。结果绘制的生长曲线表明，p38^-/-细胞的生长速率明显下降，增殖受到抑制，细胞倍增时间延长，培养96h时，p380^-/-细胞数量较p38^＋/＋减少了15．5％，说明p38基因敲除明显抑制了G2／M的进程。流式细胞仪检测结果显示，p3^＋/＋。细胞中G0／G1期细胞占34．47％，G2／M期占10．81％，S期占54．72％；p380^-/-细胞中G0／G1期占48．49％，G2／M期占4．06％，S期占47．44％。与p38^＋/＋细胞相比，G1期的p380^-/-细胞增加了40．7％，S期则减少了13．3％，说明p38基因敲除抑制了G1／S的进程。结论p38基因敲除能够阻滞细胞周期进展，减慢细胞增殖速度。Objective To investigate the effect of p38 mitogen-activated protein kinase （MAPK） gene knockout on the proliferation of embryonic fibroblasts in mice （MEFs）. Methods The expression of p38 in MEFs p38^＋/＋ and p38^-/- cells were detected by Western blotting. The growth curves of p38^＋/＋ and p38^-/- cells were plotted with the results of methylthiazoletetrazolium （MTT） colorimetric assay, and the ratios of different cell phases of p38^＋/＋ and p38^-/- cells were analyzed by flow cytometry. Results The growth curves showed that the growth rate was notably retarded and cell double time elongated in p38^-/- cells, anti＇there was 15.5% decrease of the number of p38^-/- cells in comparison with that of p38^＋/＋ cells in 96-hour culture. G2/M transition was inhibited in p38^-/- cells. Meanwhile, G1/S transition was also inhibited in p38^-/- cells, as shown by the results of flow cytometry. The ratios of G0/G1, G2/M, and S phases of p38^＋/＋ cells were 34.47%, 10.81%, and 54.72%, respectively; while those of p38^-/- cells were 48.49%, 4.06%, and 47.44%, respectively. There were 40.7% increase and 13.3% decrease in the cell numbers of G1 and S phases of p38^-/- cells in comparison with those of p38^＋/＋ cells, respectively. Conclusion p38 gene knockout in MEFs leads to ceil cycle arrest and decreased ceil proliferation.

关 键 词：P38丝裂原活化蛋白激酶 基因敲除 细胞增殖 

分 类 号：R686[医药卫生—骨科学]