目的 探讨Caspase-3、Survivin在UVB诱导HaCaT凋亡细胞中的作用.方法 研究对象为人角质形成细胞HaCaT细胞,实验分为正常对照组、10、20、40、80 mJ/cm2中波紫外线组(UVB).四甲基偶氮唑蓝(MTT)比色法观察增殖能力;流式细胞仅(FCM)实验检测细胞凋亡;实时定量PCR、Western-blot检测细胞内的Survivin、Caspase-3表达水平.结果 与正常对照组相比,NB-UVB照射组细胞增殖抑制作用及凋亡明显增强(P＜0.05),并随着照射剂量的增加,其细胞凋亡作用增加.同时与正常对照组相比,不同UVB照射剂量组Caspase-3表达均增强,其表达增强程度随着照射剂量的增加而增加,与正常对照组相比,10 rnJ/cm2照射组,细胞表达Survivin水平最高(P＜0.05),20 mJ/cm2组Survivin表达水平下降,稍低于对照组水平,40、80 mJ/cm2组Survivin进一步下降,较对照组下降明显(P＜0.05).结论 Survivin、Caspase-3参与了UVB诱导HaCaT凋亡细胞.Survivin表达水平与UVB辐射剂量有关.%Objective In order to evaluate the role of Caspase-3,Survivin in HaCaT cells apoptosis induced by NB-UVB irradiation.Methods HaCaT cells were divided into five groups,blank control group,10,20,40,80 mJ/cm2 UVB radiation groups.After the HaCaT cells were irradiated by UVB,the cell viability measured by MTT assay,and reverse transcription quantitative PCR,Western-blotting were used to detect the expression levels of antiapoptotic gene Surviving,proapoptotic gene caspease-3.Results Compared with the control group,the expression of Caspase-3 was significantly increased,but effect-dose relationship could not be seen.Meanwhile the expression of Survivin was markedly up regulated of 10 mJ/cm2 UVB radiation group,from 10 mJ/cm2 to 80 mJ/cm2 UVB radiation group,the Survivin level gradually decreased.Conclusion Caspase-3 and Survivin play an important role in HaCaT cells apoptosis induced by NB-UVB irradiation.The expression level of Survivin is related to the dose of UVB radiation.
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