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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 investigate the effects of RNA interference of bone marrow stromal cells (BMSCs) and myeloma cell line U266 APE1 on the proliferation and apoptosis of co-cultured U266 cells. Methods The APE1 siRNA expression vector was introduced into BMSCs and U266 cells respectively. The expression of APE1 protein was detected by Western blotting in two kinds of cells. Two cells were treated with APE1 siRNA, and the co-culture model of BMSCs and myeloma cells was constructed by Transwell plugged culture dishes. MTT and Annexin V-PE / 7-AAD double staining The proliferation, apoptosis and the expression of IL-6 / IL-8 mR-NA in U266 cells were detected by RT-PCR. Results APE1 siRNA could significantly decrease APE1 protein expression in BMSCs and U266 cells compared with the control group (P <0.01). In APCs co-cultured with BMSCs and U266 cells, the proliferation of U266 cells was inhibited (P <0.01). The levels of IL-6 and IL-8 mRNA in U266 cells were also decreased (P <0.01). Conclusion Inhibition of APE1 expression in BMSCs and / or myeloma cell line U266 can significantly inhibit the proliferation and promote the apoptosis of U266 cells in co-culture system.