单抗NJ001杀伤肺腺癌细胞机制的初步研究
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国家自然科学基金(81371894,81272324, 81000754,81101322,81201359);国家临床检验重点专科建设项目;江苏高校优势学科建设工程基金项目;江苏省实验诊断学重点实验室基金(XK201114)


Researches on lung adenocarcinoma cell killing mechanism of McAb NJ001
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    摘要:

    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-

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    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.

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史新惠,潘世扬,黄珮珺,娄鉴芳,张淑平,柯 星,许雨乔,黄 蕾,徐 婷,王 芳.单抗NJ001杀伤肺腺癌细胞机制的初步研究[J].南京医科大学学报(自然科学版),2014,(7):858-862

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  • 收稿日期:2013-11-29
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  • 在线发布日期: 2014-07-05
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