水飞蓟素和Fas激动性抗体CH11共同作用诱导A375-S2细胞凋亡

目的水飞蓟素和CH11共同作用诱导人黑色素瘤细胞A375-S2凋亡的分子机制。方法结晶紫法测定细胞生长抑制率;细胞凋亡的形态学观察,LDH法测定凋亡与坏死的比率;用免疫印迹法检测凋亡抑制性的去乙酰化酶SIRT1、Bc l-2家族成员(抗凋亡蛋白Bc l-2,Bc l-xL和促凋亡蛋白Bax)、细胞色素C和Caspase-3的表达。结果水飞蓟素和CH11共同作用能诱导A375-S2细胞发生凋亡;形态学观察可见凋亡小体的形成;免疫印迹法检测发现水飞蓟素和CH11共同作用的A375-S2细胞中Bax蛋白的表达增加,Bc l-2蛋白和Bc l-xL蛋白的表达降低,细胞色素C释放增加,pro-caspase-3表达量明显降低。结论水飞蓟素协同CH11能明显促进A375-S2细胞的凋亡。

Silymarin and agonistic Fas antibody, CH11, induced A375-S2 cell apoptosis through mitochondrial signal pathway

Aim To study the synergism of silymarin and CH11 on A375-S2 cell apoptosis.Methods Inhibitory effects on A375-S2 cell lines were measured by crystal violet test.Photomicroscopical observation and LDH activity test were used to assess apoptosis.SIRT1,Bcl-2 and Bcl-xL were detected by western blot analysis.Results Treatment with silymarin and CH11 significantly induced apoptosis in A375-S2 cells in a time-dependent manner.A375-S2 cells incubated with silymarin and CH11 for 12 h showed marked apoptotic changes including condensed chromatin,nuclear fragmentation and apoptotic bodies.Expressions of anti-apoptotic Bcl-2 and Bcl-xL(Bcl-2 family member) were decreased simultaneously.Expressions of apoptotic Bax and cytochrom C were increased.Consequently,Caspase-3,downstream of mitochondria signal pathway, more significantly expressed after A375-S2 cells treatment with silymarin and CH11 for 12 h.Conclusion Silymarin and CH11 synergistically induced A375-S2 cells apoptosis.