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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 cell cycle regulatory mechanism of WI-38 cells induced by tributyl hydroperoxide (t-BHP). METHODS: WI-38 cells were treated with t-BHP at intervals of 1 to 4 times after sepsis for 30 days. Cell senescence was induced. Cell senescence was observed by cell ultrastructure, cell cycle analysis and β-galactosidase staining Western blotting was used to detect the expression of CDK4, CDK2, cyclinD1, cyclinE, p21 and p16. RESULTS: The WI-38 cells exhibited senescence characteristics after treated with 10 μmol / Lt-BHP for 4 days. Cell proliferation and division stopped, cell volume increased, cell body became flat, secondary lysosomes increased, and the proportion of cells in G1 phase increased , β - galactosidase staining positive cells increased, suggesting that t-BHP can effectively induce cell senescence. After t-BHP treatment, the expressions of CDK4, CDK2 and cyclinE decreased and the expression of cyclinD1, p21 and p16 increased. Conclusion: t-BHP can induce cell senescence, and its mechanism may be related to the regulation of cell cycle through regulating the expression of molecules.