短波紫外线诱导V79细胞凋亡模型的建立

Establishment of apoptosis model in V79 cells induced by ultraviolet-C irradiation

目的探讨短波紫外线(UVC)诱导V79细胞凋亡的最佳暴露剂量并建立细胞凋亡模型。方法处于对数生长期的V79细胞,以4.5 mJ/cm^2强度UVC分别照射0、10、20、30、60、90 s,对应的照射剂量分别为0(对照)、45、90、135、270、405mJ/cm^2。采用MTT法检测细胞存活率,采用流式细胞仪检测细胞凋亡和坏死情况及细胞内活性氧(ROS)的水平。结果与对照组比较,各剂量UVC暴露组V79细胞的存活率均较低,差异有统计学意义(P<0.05);且随着UVC暴露剂量的升高,V79细胞的存活率呈下降趋势。当UVC暴露剂量为90 mJ/cm^2时,V79细胞的存活率为(50.4±5.7)%。与对照组比较,各剂量UVC暴露组V79细胞的凋亡率均较高,差异有统计学意义(P<0.05);而坏死率无明显改变。且随着UVC暴露剂量的升高,V79细胞的凋亡率呈上升趋势。与对照组比较,各剂量UVC暴露组V79细胞内ROS的水平均较高,差异有统计学意义(P<0.05);且随着UVC暴露剂量的升高,V79细胞内ROS的水平呈上升趋势。结论 UVC诱导V79细胞凋亡的最佳暴露剂量为90 mJ/cm^2。

Objective To probe the optimal exposure dose of ultraviolet-C(UVC) to induce apoptosis in V79 cells and establish the cell apoptosis model. Methods V79 cells in logarithmic growth phase were irradiated with 4.5 mJ/cm^2 UVC for0,10,20,30,60 and 90 s,the corresponding doses were 0(control),45,90,135,270,and 405 mJ/cm^2 respectively. MTT assay was used to detect cell survival rate,flow cytometry was used to detect apoptosis,necrosis and intracellular ROS. Results Compared with the control group,the survival rate of V79 cells in each UVC exposure group significantly decreased(P<0.05),and revealed downward trend with the increase of UVC exposure doses,and reached(50.4±5.7)% at 90 mJ/cm^2. Compared with the control group,the apoptosis rate of V79 cells in each UVC exposure group was higher(P<0.05),and revealed upward trend with the increases of UVC exposure doses,however no significant change was observed in the necrosis rate.Compared with the control group,the intracellular ROS level of V79 cells in each UVC exposure group significantly increased(P <0.05),and revealed upward trend with the increases of UVC exposure doses. Conclusion The optimal exposure dose for UVC to induce apoptosis of V79 cells is considered as 90 mJ/cm^2.