RNA干扰抑制KM3细胞APE1蛋白表达对其增殖与凋亡的影响

目的探讨脱嘌呤/脱嘧啶核酸内切酶(apurinic/apyrimidinic endonuclease,APE1)基因RNA干扰对KM3人多发性骨髓瘤(multiple myeloma,MM)细胞APE1蛋白表达及细胞增殖、凋亡的影响,以期为转基因治疗多发性骨髓瘤提供实验依据.方法将构建的APE1 siRNA表达载体导人人多发性骨髓瘤细胞株KM3细胞,应用Western blot检测APE1蛋白表达的改变,并通过细胞增殖曲线、MTT法和流式细胞术、DNA片段化百分率检测观察靶细胞的增殖与凋亡.结果 Western blot分析表明APE1 siRNA表达载体使KM3细胞APE1蛋白表达下降,细胞生长曲线观察、MTT法检测显示,转染的KM3细胞生长受抑,流式细胞术和DNA片段化百分率检测观察到APE1 RNA干扰使靶细胞凋亡明显增加.结论 APE1 RNA干扰通过降低APE1蛋白表达,抑制KM3细胞生长和促进其凋亡.

Proliferation and apoptosis of KM3 cells after decreasing APE1 protein level using silencing RNA

Objective To explore the relationship between the expression of APE1 protein and proliferation, apoptosis of KM_ 3 cells by decreasing apurinic/apyrimidinic endonuclease (APE1) gene expression by RNA interfering. Methods APE1siRNA expression vector or pSilencer U_6-control (as blank control) were transfected to KM_ 3 cells by liposome. The changes of APE1 protein was detected by Western blotting. The cellular proliferation capacity was observed with cell growth curve and MTT assay. The cell cycle distribution and apoptosis rate were measured by flow cytometry and DNA fragmentation rate. Results The expression of APE1 protein decreased in KM_ 3 cells by RNAi. The proliferation of transfected KM_ 3 cells was inhibited and the apoptosis of the transfected KM_ 3 cells increased significantly. Conclusion The APE1-siRNA could inhibit the proliferation and improve the apoptosis of KM_ 3 cells.