【作者】 刘奕； 于秉治；

【Author】 Liu Yi Yu BingzhiDepartment of Biochemistry, School of Basic Medical Science, China Medical University, Shenyang 110001, China, E-mail: liuyi256@126.com

【机构】 中国医科大学基础医学院生物化学研究室；

【摘要】 目的:探讨Hela细胞生长的机制。方法:用RT-PCR,Western印迹及蛋白激酶活性测定等方法对同步化的Hela细胞的细胞周期不同时相中mTOR(mammalian target of rapamycin),P70 S6激酶(P70 S6K)的α1,α2,β1,β2不同亚型及起始因子4E结合蛋白1(4EBP1)的表达进行了研究。结果:RT-PCR的结果表明:在G1,S1,G2,M1,M2几个细胞周期时相中,mTOR的mRNA的表达无明显变化。mTOR的底物P70 S6K的亚型仅α1,α2,β1,β2在M期表达均有明显增加。4EBP1的表达在M期明显减少。免疫印迹的结果与RT-PCR的一致,即M期P70 S6K的α1,α2均有增加,4EBP1在M期减少。而活性测定表明,G2期,M期mTOR较其它期有明显增加,4EBP1在M期活性有所下降。结论:mTOR,P70 S6K,4EBP1很可能在Hela细胞的生长中起重要的调节作用。更多还原

【Abstract】 Objective: In order to investigate the mechanism of cells growth. Methods: the expression of mTOR (mammalian target of rapamycin), and it’s α1, α2, β1,β2 isoforms of P70 S6 kinase(P70 S6K) and 4EBPl in defferent cell cycle phases of synchronized Hela cells were studied, by means of RT-PCR, Western-blot and protein kinase activity test. Results: The result of RT-PCR showed that the expression of mTOR mRNA had not changes obviously. In M phase, the expression isoforms of αl, α2, β1,β2 of P70 S6K, the substrates of mTOR, increase obviously, while that of 4EBP1 decreases. The results of Immunoblots were same as the RT-PCR. Activity assays showed that, in phase G2 and M, the expression of mTOR maintains at higher level than any other phase, while 4EBP1 decreases in phase M. Conclusions: The result suggested that mTOR, P70 S6K and 4EPB1 might play important roles on regulation of Hela cells growth.更多还原