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目的 探讨细胞周期 S期滞留细胞凋亡的分子基础。方法 用 Westernblot法测定 L- 2和 Br1-3pr- 1两细胞株细胞周期 S期进程相关分子 Rb、周期素 A、E、CDK2、CDC2及与细胞凋亡直接相关分子 Bcl- 2和Bax的表达 ,并用组蛋白磷酸化法测定周期素 A、E、CDK2和 CDC2相关激酶的活性。结果 经 PAL A处理后 ,发生 S期细胞凋亡的 L- 2细胞与不发生 S期细胞凋亡的 Br1- 3pr- 1细胞比较 ,其周期素 A的表达明显增加 ,Bcl- 2的表达下降。但经 PAL A处理后 ,其他分子的表达和激酶活性在上述两种细胞之间无明显差异。结论 周期素 A可能是 S期检查点基因的重要候选者 ,并可能通过调节 Bcl- 2的表达来控制 S期滞留细胞凋亡的发生。
Objective To investigate the molecular basis of cell cycle apoptosis in S phase retention cells. Methods Western Blot was used to determine the expression of Rb, cyclin A, E, CDK2, CDC2, and Bcl-2 and Bax which are related molecules in the cell cycle progression of L-2 and Br1-3pr-1 cell lines. The activity of cyclins A, E, CDK2, and CDC2 related kinases was determined by histone phosphorylation. Results After treatment with PAL A, the expression of cyclin A was significantly increased and the expression of Bcl-2 was decreased in L-2 cells with apoptosis in S phase and Br1-3pr-1 cells without apoptosis in S phase. . However, after PAL A treatment, there was no significant difference in the expression and kinase activity of other molecules between the above two types of cells. Conclusion Cyclin A may be an important candidate for S-phase checkpoint gene and may control the apoptosis of S-phase retention cells by regulating the expression of Bcl-2.