赖氨大黄酸对HeLa细胞周期的影响及其机制

Effect of Rhein Lysinate on Cell Cycle of HeLa Cell and Its Mechanism

目的探讨赖氨大黄酸对HeLa细胞周期的影响及其机制。方法 MTT法检测赖氨大黄酸对HeLa细胞增殖的影响;流式细胞技术检测活性氧水平和细胞周期变化;显微镜下观察HeLa细胞形态变化;Western blot法检测p53和p21蛋白表达。结果赖氨大黄酸剂量依赖性地抑制HeLa细胞增殖,IC50约为83μmol·L-1。随着赖氨大黄酸剂量的增加,HeLa细胞中活性氧水平增加,并出现S期和G2期阻滞,以及细胞空泡样变性。通过Western blot研究发现赖氨大黄酸处理后的p53蛋白磷酸化水平呈剂量依赖性地增加,同时下游周期相关蛋白p21表达水平也随之增加,而p53蛋白表达水平未出现显著地剂量依赖性变化。结论赖氨大黄酸通过增加HeLa细胞中活性氧水平,引起细胞周期相关蛋白p53的磷酸化水平和p21蛋白表达水平增加,引起HeLa细胞出现空泡样变性,并诱导S期和G2期阻滞。

OBJECTIVE To explore the effect of Rhein lysinate on cell cycle of HeLa cells and its mechanism.METHODS The effect of Rhein lysinate on cell proliferation was detected by MTT.The level of reactive oxygen species and the change of cell cycle of HeLa cells were investigated by flow cytometry.Cell morphology was observed by microscope.The expression of p53 and p21 was determined by Western blot.RESULTS Rhein lysinate could inhibit the proliferation of HeLa cells in a dose-dependent manner.The IC50 was approximately 83 μmol·L-1.The level of reactive oxygen species and the ratios of S period and G2 period cells were increased following the increase of Rhein lysinate.Moreover vacuolar degeneration was induced with the increasing of Rhein lysinate.The phosphorylation of p53 and the expression of p21 were also increased.However the expression of p53 was not increased in accordance with the increase of Rhein lysinate.CONCLUSION Rhein lysinate inhibits the proliferation of HeLa cells by increasing the levels of reactive oxygen species,improving the phosphorylation of p53 and the expression of p21,presenting vacuolar degeneration and inducing the S period and G2 period block.