RNA干扰骨髓基质细胞和/或骨髓瘤细胞APE1表达对共培养骨髓瘤细胞增殖及凋亡的影响  

Effects of RNA interference inhibiting APE1 expression in bone marrow stromal cells and/or U266 cells on proliferation and apoptosis of co-cultured U266

作　　者：谢家印[1] 杜佳[1] 李梦侠[1] 卿毅[1] 关伟[1] 曾林立[1] 王东[1] 

机构地区：[1]第三军医大学大坪医院野战外科研究所肿瘤中心,重庆400042

出　　处：《第三军医大学学报》2011年第10期1008-1011,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(30772520)~~

摘　　要：目的探讨RNA干扰骨髓基质细胞(bone marrow stromal cells,BMSCs)及骨髓瘤细胞株U266 APE1表达对共培养的U266细胞增殖、凋亡的影响。方法将构建的APE1 siRNA表达载体分别导入BMSCs及U266细胞中。Westernblot法检测2种细胞中APE1蛋白表达;2株细胞经APE1 siRNA处理后采用Transwell插入式培养皿构建BMSCs与骨髓瘤细胞共培养模型,采用MTT法、Annexin V-PE/7-AAD双染法、RT-PCR分别检测U266细胞增殖、凋亡及细胞中IL-6/IL-8 mR-NA表达水平。结果 APE1 siRNA可明显降低BMSCs及U266细胞APE1蛋白表达,与对照组相比差异具有统计学意义(P<0.01);在APE1 siRNA同时处理BMSCs和U266细胞后的共培养体系中,U266细胞增殖抑制及细胞凋亡明显高于单一细胞APE1敲低组及APE1 siRNA未处理组(P<0.01);U266细胞中IL-6及IL-8 mRNA表达水平亦降低(P<0.01)。结论抑制APE1在BMSCs和/或骨髓瘤细胞株U266的表达,可明显抑制共培养体系中U266细胞的增殖活性并促进其凋亡。Objective To study the effect of RNA interference inhibiting apurinic/apyrimidinic endonuclease 1（APE1） expression in bone marrow stromal cells（BMSCs） and myeloma cell line U266 on the proliferation and apoptosis of co-cultured U266 cells.Methods Constructed APE1 siRNA expression vector Ad5v-APE1 siRNA was used to transfect BMSCs and U266 cells.A co-culture system of BMSCs and U266 cells was established with Transwell insert culture dish.The co-culture system of BMSCs and U266 cells were divided into four groups： a control group（untransfected BMSCs and U266 cells）,a first single treatment group（transfected BMSCs and untransfected U266 cells）,a second single treatment group（untransfected BMSCs and transfected U266 cells）,and a dual treatment group（transfected BMSCs and U266 cells）.The expression levels of APE1 were detected by Western blotting.The proliferation,apoptosis,and mRNA expression levels of interleukin-6（IL-6） and interleukin-8（IL-8） of co-cultured U266 cells were examined by MTT assay,flow cytometry with annexin V-PE/7-AAD double staining,and RT-PCR,respectively.Results BMSCs and U266 cells transfected with APE1 siRNA had significantly lower protein expression of APE1 than untransfected cells（P0.01）.Compared with the two single treatment groups and the control group,the proliferation inhibition and apoptosis degrees of U266 cells in the dual treatment group significantly increased（P0.01）.Compared with the control group,the mRNA expression levels of IL-6 and IL-8 in U266 cells significantly decreased in the two single treatment groups（P0.01） and the dual treatment group（P0.01）,showing greater decrease in the dual treatment group（P0.01）.Conclusion Inhibiting APE1 expression in BMSCs and/or U266 cells in a co-culture system suppresses the proliferation and promotes the apoptosis of U266 cells.

关 键 词：多发性骨髓瘤 细胞凋亡 APE1基因 RNA干扰 

分 类 号：R733.3[医药卫生—肿瘤] R73-351[医药卫生—临床医学]