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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。
Objective: To observe the activity of monoclonal antibody NJ001 against lung adenocarcinoma cells and explore its molecular mechanism. Methods: The lung adenocarcinoma cell line SPC-A1 was selected as the research object. The apoptotic rate was detected by flow cytometry. The morphological changes of apoptotic cells were observed by phase contrast microscopy. The expressions of caspase-8, caspase-9 , Caspase-3, PARP and Bcl-2, Bax expression changes. Results: The monoclonal antibody NJ001 could induce apoptosis of SPC-A1 cells in a dose-dependent manner. SPC-A1 cells were observed under microscope to shrink round, even fall off, showing typical changes in apoptosis; Western blot results showed that in the monoclonal antibody NJ001, PARP protein was cut, the expression of caspase-3 was Cleaved and decreased, caspase-8 and caspase-9 protein showed activation, the expression of cleaved caspase-8 and cleaved caspase-9 protein expression. In addition, the pro-apoptotic protein Bax was up-regulated and the anti-apoptotic protein Bcl-2 was down-regulated. CONCLUSION: The monoclonal antibody NJ001 exerts its activity of killing lung adenocarcinoma cells by inducing apoptosis, and the mechanism of apoptosis is related to the activation of caspase and the regulation of Bcl-2 family proteins.