摘要
目的观察抗CD44单克隆抗体(mAb)A3D8对腹水源卵巢癌球形体形成细胞(A-SFC)凋亡的影响,并探讨其机制。方法将常规培养的A-SFC随机分为观察组和对照组,观察组加入A3D8至其终浓度为2 mg/L,对照组不加药。培养24 h后用Annexin V-FITC/PI凋亡检测试剂盒检测细胞凋亡率;用罗丹明123试剂盒测算细胞线粒体膜电位;用Western blot法检测A-SFC细胞中的CDK2、cyclin A、Bcl-2和Caspase-3。结果观察组细胞凋亡率11.62%±1.01%、线粒体膜电位损失率31.32%±3.47%,对照组分别为6.02%±0.79%、17.65%±2.03%,两组细胞凋亡率和线粒体膜电位损失率相比P均<0.05。观察组CDK2、cyclin A、Bcl-2、Caspase-3蛋白相对表达量分别为对照组的79%、64%、95%、160%。结论 A3D8可促进A-SFC凋亡,其机制可能与A3D8下调CDK2、cyclin A、Bcl-2表达,上调Caspase-3蛋白表达,促进线粒体膜电位损失有关。
Objective To explore the effects and mechanisms of A3D8（mAb against CD44） on apoptosis in sphere-forming cells from ascites（A-SFC） of ovarian cancer.Methods The routine cultured A-SFC cells were divided into observation and control group,the observation group was put into 2 mg/L A3D8,and control group nothing.After the 24 h cultivation,Annexin V-FITC/PI apoptosis detection kit was used to analyze cell apoptosis.Rhodamine123 apoptosis detection kit was used to detect the change of mitochondrial transmembrane potential（ΔψM）.Western blot assay was used to detect the CDK2,cyclin A,Bcl-2 and Caspase-3 protein levels.Results The apoptosis rates of observation group was 11.62%±1.01%,the loss of ΔψM was 31.32%±3.47%,and the control group was 6.02%±0.79%、17.65%±2.03%.Compared to control group,P〈0.05 all.The CDK2,cyclin A,Bcl-2 and Caspase-3 relative protein level in obsenation group was 79%,64%,95%,160% compared with control group（P〈0.05 all）.Conclusion A3D8 can cause apoptosis on A-SFC,the probably mechanism maybe that A3D8 reduce CDK2、cyclinA、Bcl-2 level,raise Caspase-3,raise the loss of ΔψM.
出处
《山东医药》
CAS
2012年第6期5-7,共3页
Shandong Medical Journal
基金
国家自然科学基金资助项目(30670801)
天津市科学技术委员会基金资助项目(06YFJMJC08300)
关键词
A3D8
腹水源卵巢癌球形体形成细胞
细胞凋亡
anti-CD44 monoclonal antibody A3D8 sphere-forming cells from ascites of ovarian cancer apoptosis
