人白细胞介素1β通过线粒体途径激活caspase-3诱导A375-S2细胞凋亡  

Interleukin-1β induces apoptosis in human melanoma A375-S2 cells through mitochondrial pathway by activating caspase-3

作　　者：王澈[1] 王敏伟[1] 田代真一[2] 小野寺敏 池岛乔[1] 

机构地区：[1]沈阳药科大学中日医学药学研究所 [2]昭和药科大学病态科学教研室

出　　处：《免疫学杂志》2005年第5期403-405,408,共4页Immunological Journal

摘　　要：目的研究人白细胞介素1β(Interleukin-1β,IL-1β)体外诱导人黑色素瘤A375-S2细胞凋亡的机制。方法通过Hoechst33258荧光染色,观察A375-S2细胞在IL-1β作用下的形态学变化。使用琼脂糖凝胶电泳法检测IL-1β对细胞DNA降解的影响。应用caspase活力测定法检测IL-1β对caspase-3活力的影响。利用Westernblot方法检测IL-1β对caspase-3底物的降解以及对细胞内Bcl-2家族和AIF蛋白表达的调节作用。结果IL-1β(1nmol/L)能够诱导A375-S2细胞凋亡,72h时细胞体变小,细胞核呈现固缩、断裂,并出现因凋亡引起的DNA梯状条带。IL-1β诱导细胞凋亡过程中,caspase-3的活力升高,其两种底物PARP和ICAD均发生降解,线粒体中抗凋亡蛋白Bcl-2和Bcl-xL的表达减少,促凋亡蛋白Bax的表达增加。凋亡诱导因子AIF蛋白的表达也有所增加。结论IL-1β通过激活caspase-3降解其底物,激活AIF,并上调线粒体内Bax/Bcl-2和Bax/Bcl-xL的表达比例诱导A375-S2细胞凋亡。Objective To investigate the mechanism of IL-1β-induced cell apoptosis in human malignant melanoma A375-S2 cells. Methods The morphologic changes of cell were observed by using Hoechst 33258 staining. DNA fragmentation was detected by agarose gel electrophoresis. Caspase-3 activity was assessed by assaying the activity of caspase. The degradation of caspase-3 substrates and the protein expression of DBcl-2 family and AIF were assayed by Western blotting. Results Cell nuclear condensation was observed after treatment with IL-1β for 48h and DNA fragmentation was observed at 72h. In apoptotic progress induced by IL-1β, caspase-3 activation was augmented and its two substrates, PARP and ICAD proteins, were degradation. Expressions of antiapoptotic Bcl-2 and Bcl-xL were decreased, but proapoptotic Bax expression was slightly increased. Furthermore, AIF expression was up-regnlated. Conclusion IL-1β induces apoptosis in A375-S2 cells by activating caspase-3 and AIF, as well as up-regnlating the ratio of the Bax/Bcl-2 expression to Bax/Bcl-xL expression.

关 键 词：白细胞介素1Β 人黑色素瘤 凋亡 CASPASE-3 BCL-2 

分 类 号：R73-3[医药卫生—肿瘤]