抗CD44mAbA3D8对卵巢癌球形体形成细胞增殖和凋亡的影响  

Influence of Anti-CD44 mAb A3D8 on Proliferation and Apoptosis in three Sphere-Forming Cells of Ovarian Cancer

作　　者：许静静[1] 杜涌瑞[1] 张秋月[1] 顾欣[1] 谷超[1] 李凤舞[1] 邓为民[1] 

机构地区：[1]天津医科大学基础医学院免疫学教研室,300070

出　　处：《天津医药》2012年第7期641-644,共4页Tianjin Medical Journal

基　　金：国家自然科学基金资助项目(项目编号:30670801);天津市科学技术委员会基金资助项目(项目编号:06YFJMJC08300)

摘　　要：目的:探讨抗CD44单克隆抗体(mAb)A3D8对3种卵巢癌球形体形成细胞增殖和凋亡的影响,并探讨其作用机制。方法:用MTS法检测A3D8对细胞增殖的影响;采用PI染色及流式细胞仪检测A3D8对细胞周期的影响;用AnnexinV-FITC/PI凋亡检测试剂盒检测A3D8在细胞凋亡中的作用;用罗丹明123试剂盒检测A3D8对细胞线粒体膜电位的影响;并采用WesternBlotting法检测A3D8作用于3种卵巢癌球形体形成细胞后,CDK2、cyclinA、Bcl-2和caspase-3的改变。结果:A3D8可抑制3种卵巢癌球形体形成细胞的增殖,且此抑制作用呈现剂量和时间依赖性;A3D8可在阻滞S期的同时降低G0/G1期比例;A3D8可促进3种细胞凋亡,与顺铂(DDP)联用,细胞凋亡率较单独使用DDP增高;A3D8的处理可导致3种细胞的线粒体膜电位损失,CDK2、cyclinA、Bcl-2蛋白表达下调,caspase-3表达上调。结论:A3D8可能是通过影响p21/CDK2/cyclinA途径阻滞细胞周期达到抑制3种卵巢癌球形体形成细胞增殖的结果,并且可通过线粒体途径促进细胞凋亡。Objective： To explore the effects and mechanisms of A3D8 （mAb） against CD44 on proliferation and apoptosis in three kinds of sphere-forming cells of ovarian cancer. Methods： The MTS assay was used to evaluate the effect of A3D8 on the cell proliferation. The PI staining and flow cytometry was used to analyze the effect of A3D8 on the cell cycle distribution. Annexin V-FITC/PI apoptosis detection kit was used to analyze the effect of A3D8 on the cell apoptosis. Rhodamine123 apoptosis detection kit was used to detect the effect of A3D8 on the change of mitochondrial transmembrane potential （△ψM）. Western blotting assay was used to detect the effect of A3D8 on the CDK2, cyclinA, Bcl-2 and caspase-3 protein levels in three kinds of sphere-forming cells of ovarian cancer. Results： A3D8 inhibited the proliferation in three sphere-forming cells of ovarian cancer, which showed a concentration- and time-dependant manner. A3D8 caused S arrest with GO/G1 decrease via p21/CDK2/cyclinA pathways. A3D8 promoted apoptosis rates of three kinds of cells. The apoptosis rates were higher in cells treated by A3D8 combined with DDP than those of DDP treatment alone. A3D8 caused AtOM to decrease, and decreased CDK2, cyclinA and Bcl-2 protein levels, but increased caspase-3 expression in three kinds of sphere-forming cells. Conclusion： A3D8 inhibited the proliferation in three sphere-forming cells of ovarian cancer probably by the cell cycle arrest via p21/CDK2/cyclin A pathways A3D8 increased apoptosis rates via mitochondrial pathways.

关 键 词：抗原 CD44抗体 单克隆 卵巢肿瘤 细胞增殖 细胞凋亡 卵巢癌球形体形成细胞 

分 类 号：R733.71[医药卫生—肿瘤]