单抗NJ001杀伤肺腺癌细胞机制的初步研究

目的:观察单克隆抗体NJ001杀伤肺腺癌细胞的活性,并探索其分子机制?方法:选择肺腺癌细胞株SPC-A1作为研究对象,流式细胞仪检测细胞凋亡率;相差显微镜观察凋亡细胞形态学改变;Western blot检测细胞凋亡相关蛋白caspase-8?caspase-9?caspase-3?PARP和Bcl-2?Bax表达的变化?结果:单克隆抗体NJ001可以导致SPC-A1细胞发生凋亡,且凋亡率呈剂量依赖性?显微镜下观察到SPC-A1细胞皱缩变圆,甚至脱落,呈现典型的凋亡改变;Western blot检测结果显示,在单抗NJ001作用下,PARP蛋白被剪切,caspase-3的表达量因被剪切而降低,caspase-8和caspase-9蛋白均表现活化,表现为cleaved caspase-8和cleaved caspase-9蛋白的表达上调?另外促凋亡蛋白Bax表达上调,抗凋亡蛋白Bcl-2的表达下调?结论:单克隆抗体NJ001能通过诱导细胞凋亡发挥其杀伤肺腺癌细胞的活性,且其凋亡机制与caspase活化和Bcl-2家族蛋白的调节有关?

Researches on lung adenocarcinoma cell killing mechanism of McAb NJ001

Objective:To observe the anti-cancer activity of McAb NJ001 against lung adenocarcinoma cells,and explore its molecular mechanism. Methods:We selected SPC-A1 cell lines as research object,used flow cytometry to detect cell apoptosis,and observed morphological changes of SPC-A1 cells by phase contrast microscope. We detected the expressions of apoptosis-related proteins such as caspase-8,caspase-9,caspase-3,PARP,Bax,and Bcl-2 by Western blotting assays. Results:McAb NJ001 induced apoptosis in SPC-A1 cells in a dose-dependent manner. After treatment with NJ001,SPC-A1 cells showed shrinkage,and became round or even fell off,which were typical morphological changes of apoptosis. Results:Western blots showed that after treatment with McAb NJ001,PAPR were cleaved so that caspase-3 expression was decreased,caspase-8 and caspase-9 were activated and their protein expression levels were increased. The expression of pro-apoptotic protein Bax increased while the expression of anti-apoptotic protein Bcl-2 decreased. Conclusion:McAb NJ001 has strong anti-cancer activity due to its apoptosis inducing effect. In addition,caspase activation and up-regulation of Bax/Bcl-2 were involved in NJ001-induced apoptosis.