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钙调素拮抗剂对HeLa细胞S期进程的影响 |
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孙 霞 柳惠图 童迎凯 王端顺
【摘要】 目的 探讨钙调素拮抗剂三氟拉嗪(TFP)对HeLa细胞S期进程的影响及其分子机理。 方法 通过TdR双阻断法获得同步化的S期HeLa细胞，以3H-TdR掺入法和Western技术分别观察了TFP对HeLa细胞S期进程和胸苷激酶(thymidine kinase,TK)活性及细胞周期引擎分子CyclinA、Cdk2和细胞周期调节蛋白p80cdc25B、PCNA、p21蛋白表达水平的影响。 结果 TFP(20μmol/L)能使3H-TdR的掺入峰值后移，并抑制了CyclinA、PCNA、p80cdc25B的表达和提高了p21蛋白的水平，但对Cdk2的表达几乎无影响。 结论 CaM除了能通过影响PCNA的水平直接作用于DNA合成，同时又可通过作用于CyclinA、p80cdc25B、p21等周期引擎分子和调控因子水平正调HeLa细胞S期进程。
【关键词】 钙调素 三氟拉嗪 S期 HeLa细胞
THE EFFECTS OF ANTAGONIST OF CaM ON THE PROGRESSION
OF S PHASE IN HeLa CELLS
Sun Xia, Liu HuituΔ, Tong Yingkai, Wang Duanshun
(Key Laboratory of Cell Proliferation and Regulation,Department of Biology,Beijing Normal University,Beijing)
【Abstract】 Objective To study the effects of antagonist of CaM(TFP) on the progression of S phase and the molecular mechanism in HeLa cells. Methods The synchronized S phase cells were obtained by the method of thymidine(TdR)double block.The progression of S phase and the activity of thymidine kinase were determined through 3H-TdR incorporation assay.Western blotting was used to analyze the levels of CyclinA,Cdk2,PCNA,p80cdc25B and p21 proteins. Results The TFP(20μmol/L)put off the appearing of the peak of 3H-TdR incorporation and inhibited S phase progression.The activity of thymidine kinase decreased after treatment with TFP.The TFP(20μmol/L)decreased the level of CyclinA,PCNA，p80cdc25B increased the expression of p21 protein,but not affected on the expression of Cdk2. Conclusion CaM could positively regulate the S phase progression in HeLa cells not only by affecting on the synthesis of DNA through PCNA,but also by influencing the expression of cell cycle engine molecules and related regulators including CyclinA,p80cdc25B,and p21 proteins.This may be one of the molecular mechanisms which is involved in the positive regulation of S phase progression in HeLa cells by CaM.
【Key words】 Calmodulin(CaM); Trifluoper[1] [2] [3] 下一页
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