目的:探讨熊果酸是否能够通过核因子(NF)-κB信号通路对皮肤鳞状细胞癌(以下简称鳞癌)细胞增殖、凋亡产生影响.方法:以人皮肤鳞癌细胞株A431细胞为研究对象,人永生化角质形成细胞系HaCaT细胞为正常对照,氟尿嘧啶(5-FU)为药物阳性对照,传代培养皮肤鳞癌细胞株A431细胞,不同浓度熊果酸作用于细胞后,四甲基偶氮唑蓝(MTT)法观察熊果酸对细胞增殖的影响,流式细胞仪检测细胞凋亡率,hoechst染色检测细胞凋亡形态,Western blot检测IκBαt、pIκBα、bc1-2、c-IAP2等NF-κB信号通路相关蛋白的表达.结果:与对照组比较,不同浓度熊果酸组均出现细胞皱缩及细胞破碎现象,细胞凋亡率增加,并呈剂量依赖性,pIκBα、bc1-2、c-IAP2蛋白表达下降,IκBα蛋白表达升高.结论:熊果酸能通过阻断NF-κB通路诱导A431细胞凋亡.
Objective:To investigate whether ursolic acid could induce apoptosis of human skin cancer cell line A431 through inhibition of NF-KB signaling pathway. Methods:A431 cells were used as the target of the study, HaCaT cells served as normal control, and 5-FU as the drug positive control. The number of viable A431 cells were analyzed using MT'F. Apoptosis of A431 cells were analyzed using flow cytometry and hoechst staining. Expression levels of IKBo~, pIKBct, bcl-2 andc-IAP2 were detected using Western blot after incubation of A431 cells with ursolic acid in different concentrations and at different time points. Results:Compared with the control group, cell shrinkage and cell broken phenomenon presented in all ursolic acid treated groups. In ursolic acid treated groups the apoptosis rates and IxBa protein expression were increased in a dose-dependent manner, but pIkBa, bcl-2, c-IAP2 protein expression were decreased. Conclusion:ursolic acid could induce apoptosis of A431 cells through inhibition of NF-kB signaling pathway.