ABT-263增敏Lexatumumab诱导三阴性乳腺癌细胞凋亡的研究

The study on ABT-263 sensitizes Lexatumumab-induced apoptosis in triple negative breast cancer cells

目的:研究Bcl-2抑制剂对于死亡受体单克隆抗体Lexatumumab激活诱导三阴性乳腺癌细胞凋亡的增敏效果。方法:体外培养ER、PR和HER-2三阴性细胞株。分为对照组、ABT-263处理组、Lexatumumab处理组、ABT-263+Lexatumumab处理组。用亚致死剂量的ABT-263,或者Lexatumumab,或者二者同时处理细胞,荧光显微镜观察对照组和处理组细胞内细胞核凋亡形态变化情况。然后凋亡细胞计数,统计凋亡率。免疫印迹(Western blotting)法检测细胞凋亡标志性蛋白Caspase 3和PARP切割条带在各组中的表达,以评估Bcl-2抑制作用对死亡受体信号通路诱导细胞凋亡的影响。结果:在荧光显微镜下,可以观察到ABT-263和Lexatumumab单独处理细胞不能够诱导三阴性乳腺癌细胞核出现固缩凝聚现象。抑制剂ABT-263能够显著逆转Lexatumumab诱导的乳腺癌细胞凋亡。结论:Bcl-2抑制剂ABT-263能够增敏单克隆抗体Lexatumumab诱导三阴性乳腺癌细胞株MDA-MB-231细胞凋亡。ABT-263和Lexatumumab联合诱导的凋亡作用具有Caspase依赖性。

Objective: To investigate the roles of Bcl- 2 inhibition in Lexatumumab- induced apoptosis in triple negative breast cancer cells. Methods: Triple negative breast cancer cells MDA- MB- 231 were cultured in vitro,to set up control group,ABT- 263- treated group,Lexatumumab- treated group and ABT- 263 plus Lexatumumab-treated group. Cells untreated or treated with various conditions were observed under microscope to show DNA fragmentation. Western blotting was performed to detect apoptotic marker molecule Caspase 3 cleavage activation and PARP cleavage. Results: Low concentrations of ABT- 263 and Lexatumumab did not induces DNA fragmentation in MDA- MB- 231 cells. Inhibition of Bcl- 2 activation by sub lethal ABT- 263 significantly sensitizes Lexatumumab- induced apoptosis in MDA- MB- 231 cells. Conclusion: Bcl- 2 inhibitor ABT- 263 is able to sensitize monoclonal antibody Lexatumumab- induced Caspase- dependent apoptosis in triple negative breast cancer cells.