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目的探讨低剂量长期砷暴露对人皮肤角质形成细胞系HaCat细胞周期及周期蛋白D1(cyclin D1)和p21影响。方法 HaCat细胞暴露于浓度为0、0.05、0.1μmol/L的NaAsO215周后,用流式细胞仪测定10 000个细胞的细胞周期分布;用western blot法检测细胞cyclin D1和p21的蛋白表达水平。结果与对照组比较,NaAsO2染毒组细胞的G0/G1期细胞数明显增高,S期细胞数明显降低,而G2/M期无明显变化;0.05、0.1μmol/L砷染毒组细胞内cyclin D1表达水平分别为(152.40±8.17)%、(145.59±2.89)%,与对照组[(99.99±1.13)%]比较,均明显增高;0.05、0.1μmol/L砷染毒组细胞内p21表达水平分别为(64.06±1.62)%、(39.57±1.82)%,均明显低于对照组的(99.9±2.83)%,呈剂量效应关系(P<0.05)。结论长期低剂量砷暴露可诱导HaCat细胞周期异常,cyclin D1表达上升,p21表达下降。
Objective To investigate the effects of low-dose long-term arsenic exposure on the cell cycle of human skin keratinocyte cell line HaCat and the expression of cyclin D1 and p21. Methods After HaCat cells were exposed to NaAsO2 at 0,0.05 and 0.1 μmol / L for 15 weeks, the cell cycle distribution of 10 000 cells was determined by flow cytometry. The protein expression of cyclin D1 and p21 was detected by western blot. Results Compared with the control group, the number of cells in G0 / G1 phase in NaAsO2-treated group was significantly increased, the number of cells in S phase was significantly decreased, while there was no significant change in G2 / M phase. In the 0.05, 0.1μmol / L arsenite- (152.40 ± 8.17)% and (145.59 ± 2.89)%, respectively, were significantly higher than those in the control group [(99.99 ± 1.13)%] (64.06 ± 1.62)% and (39.57 ± 1.82)%, respectively, which were significantly lower than that of the control group (99.9 ± 2.83)%, showing a dose-response relationship (P <0.05). Conclusion Long-term exposure to low doses of arsenic can induce an abnormal cell cycle in HaCat cells. The expression of cyclin D1 is up-regulated and the expression of p21 is decreased.