摘要
目的 :探讨三丁基过氧化氢 (t-BHP)诱导WI - 38细胞衰老的细胞周期调控机制。方法 :从 30代开始 ,隔代用t-BHP作用WI- 38细胞 4次 ,每次 1h,诱导细胞衰老 ,从细胞超微结构、细胞周期分析和β -半乳糖苷酶细胞化学染色观察衰老细胞的特点 ,同时用Westernblotting方法检测细胞周期调控蛋白CDK4、CDK2、cyclinD1、cyclinE、p2 1和p16的表达程度。结果 :10 0 μmol/Lt-BHP作用 4次后 ,WI- 38细胞出现衰老的特征 ,细胞增殖分裂停止 ,细胞体积增大、胞体变平、次级溶酶体增多 ,同时G1期细胞比例增加 ,β -半乳糖苷酶染色阳性细胞数增加 ,提示t-BHP能有效地诱导细胞衰老。t-BHP作用后CDK4、CDK2、cyclinE表达下降 ,cyclinD1、p2 1和p16表达增加。结论 :t-BHP有诱导细胞衰老的作用 ,其机制可能与通过调节细胞周期调控分子的表达有关。
AIM: To investigate the mechanism of cell c ycle in aging model induced by tert-butylhydroperoxide(t-BHP).METHODS: From the 30th population doubling(PD), WI-38 cell s were exposed for 1 h to t-BHP at every two PDs and four stresses were induced. The cell morphology and ultrastructure, cell cycle assay and β- galactosidase cytochemistry staining were used to evaluate cell senescence. Then the levels of CDK4, CDK2, cyclin D1, cyclin E, p21 and p16 protein were detecte d by Western blotting.RESULTS: After four times treated with 100 μmol/L t-BHP, the pr oliferation and division in WI-38 cells were ceased, which showed that cell beca me larger and flat, the number of secondary lysosome and activity of SA-β-galac tosidase were increased. In addition, the percentage of cells in the G 1 phase was decreased. Furthermore, it showed that the protein levels of CDK4, CDK2 and cyclin E was reduced and cyclin D1, p21 and p16 was elevated.CONCLUSION: t-BHP induces senescence in WI-38 cells. The possibl e mechanism is that t-BHP can change the expression of proteins related with cel l cycle.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2005年第2期238-242,共5页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目 (No .30 4 782 188)
福建省自然科学基金资助项目 (No.C0 2 10 0 17)
