SU11248干预白血病细胞K562对WWOX及相关基因表达的影响

Effect of SU11248 on the expression of WWOX and related protein in leukemia cell line K562

目的:探讨SU11248对白血病细胞K562的作用及对K562表达WWOX和相关蛋白的影响。方法:以MTT法检测SU11248对K562细胞增殖能力的影响;RT-PCR检测SU11248干预K562细胞前后WWOX、BCL-2、BAX mRNA表达变化。Western blot检测SU11248处理K562细胞前后Wwox、Bcl-2蛋白的表达变化。结果:SU11248作用后K562细胞增殖受到明显抑制(P<0.05),呈现剂量和时间依赖性,半数抑制浓度(IC50)约为3.2μg/ml。3.2μg/ml SU11248作用后,K562细胞BCL-2 mRNA表达水平随时间延长而逐渐降低,WWOX mRNA逐渐增强,而BAX mRNA表达无明显变化。Western blot显示Bcl-2蛋白呈时间依赖性降低,而Wwox蛋白呈时间依赖性增强。WWOX与BCL-2成负相关(P<0.05)。结论:SU11248抑制K562细胞增殖及诱导其凋亡作用可能与上调WWOX、下调Bcl-2的表达有关。

Objective：To study the effect of SUl1248 on K562 cells and the change of WWOX and related protein in K562 cells treated with SUl1248. Methods： Effect of SUl1248 on proliferation of K562 cells was detected by MTT assay. Changes of WWOX, BCL-2, BAX mRNA expression were estimated by RT-PCR. Expression of Wwox, Bcl-2 protein in K562 cells were detected by Western blot analysis. Results： The proliferation of K562 cells could be inhibited by SUl1248 in doseand time-dependent manner （ P 〈 0.05 ）. The concentration of 50% growth inhibition （ ICs0 ） of SU11248 for K562 cells was 3.2 μg/ml. After treating K562 cells with 3.2 μg/ml SU11248 at different time points, the expression of BCL-2 mRNA and protein was reduced significantly, and the ex- pression of WWOX mRNA and protein was increased significantly in a time-dependent manner （ P 〈 0.05 ）, but the expression of BAX mRNA had no change. The expression of WWOX mRNA and protein were negatively correlated to the expression of BCL-2 mRNA and protein, （P 〈 0.05 ）. Conclusion： SU11248 could inhibit the proliferation of K562 cells which may be related to down-regulation of BCL-2 and up-regulation of WWOX.