维生素E琥珀酸酯增强DR5抗体对Raji和K562细胞的凋亡作用  

Enhancement of the Apoptotic Effects of DR5 Monoclonal Antibodies by Vitamin E Succinate in Raji and K562 Cells

作　　者：李淑莲[1] 王赟[1] 都景芳[1] 马远方[1] 

机构地区：[1]河南大学细胞与分子免疫学实验室免疫学研究所,河南省开封市475004

出　　处：《中国肿瘤临床》2011年第15期878-881,共4页Chinese Journal of Clinical Oncology

基　　金：河南省杰出人才创新基金项目(编号:0321001800)资助~~

摘　　要：目的:探讨VES增强抗DR5单抗mDRA-6诱导白血病细胞凋亡作用及可能机制。方法:MTT法检测VES及mDRA-6对白血病细胞Raji和K562细胞的生长抑制作用;Annexin V-FITC/PI双染分析白血病细胞凋亡;流式细胞术检测细胞表面DR5表达;免疫印迹技术检测细胞DR5蛋白表达。结果:MTT检测表明,浓度为10μg/mL的mDRA-6作用12h,Raji及K562细胞死亡率分别为21.98%和5.23%,而5μmol/L、10μmol/L、20μmol/L的VES分别与mDRA-6共同作用于Raji或K562细胞,Raji细胞死亡率分别增加至24.67%(P>0.05)、35.65%(P<0.01)和40.22%(P<0.01),K562细胞死亡率分别增加至6.00%(P>0.05)、7.89%(P<0.01)、8.67%(P<0.01)。Annexin V/PI双染检测2μg/mL的mDRA-6单独作用Raji及K562细胞12h,细胞凋亡率分别为20.79%和7.74%,2μg/mL的mDRA-6与10μmol/L的VES共同作用Raji及K562细胞12h,细胞凋亡率分别增至43.18%和16.99%。Raji和K562细胞表面DR5自然表达分别为42.35%和14.92%;10μmol/L的VES作用Raji、K562细胞12h后,细胞膜DR5表达率分别增加至70.08%和16.38%。免疫印迹技术检测显示,不同浓度的VES均可增加Raji和K562细胞DR5蛋白表达。结论:VES增强mDRA-6对白血病细胞系Raji和K562细胞的凋亡诱导作用,可能是VES增加了Raji和K652细胞膜DR5表达及细胞DR5蛋白表达所致。Objective： To study the synergistic effect of mDRA-6 and vitamin E succinate （VES） on Raji and K562 leukemic cells and its possible mechanism. Methods： Methyl thiazolyl tetrazolium （ MTT ） assay was used to detect the growth inhibition of VES and mDRA-6 on leukemic Raji and K562 cells. AnnexinV-FITC/PI double staining was carried to analyze the apoptosis of Raji and K562 cells. Flow cytometry was performed to determine death receptor 5 （ DR5 ） expression on cell surface. An immunoblot assay was used to determine the DR5 total protein expression in leukemic cells. Results： The MTT assay showed that the specific cytotoxici- ty of mDRA-6 against Raji and K562 cells at 10 mg/L for 12 h, reached 21.98% and 5.23%, respectively. At 12 h after the 10 μg/mL mDRA-6 treatment, the death rates of the Raji and K562 cells were 21.98% and 5.23%, respectively. When mDRA-6 with VES at 5, 10, and 20 μmol/L were used to treat the Raji and K562 cells, the death rates of the Raji cells increased to 24.67% （ P 〉 0.05 ）, 35.65% （ P〈 0.01 ）, and 40.22% （ P〈 0.01 ）, respectively, and those of the K562 cells increased to 6.00% （ P〉 0.05 ）, 7.89% （ P〈 0.01 ）, and 8.67% （ P 〈 0.01 ）, respectively. Annexin V/PI double staining was used to detect the solitary effect of 2 gg/mL mDRA-6 on the Raji and K562 cells at 12 h after the treatment. The apoptotic rates of the Raji and K562 cells were 20.79% and 7.74%, respectively. At 12 h after treatment with 2 gg/mL mDRA-6 and 10 μmol/L VES, the apoptotic rates of the Raji and K562 cells reached 43.18% and 16.99%, respectively. The natural expression of DR5 on the surface of the Raji and K562 cells reached 42.35% and 14.92%, respectively. At 12 h after the effect of 10μmol/L VES on Raji and K562 cells, the DR5 expression levels at the cell membrane increased to 70.08% and 16.38%, respectively. The immunoblot assay showed that different VES concentrations could increase DR5 protein expression on Raji and K562 cells. Conclusion： VES enhances th

关 键 词：肿瘤坏死因子相关凋亡诱导配体 死亡受体5 单克隆抗体 凋亡 维生素E琥珀酸酯 

分 类 号：R733.7[医药卫生—肿瘤]