SU11248对骨髓瘤细胞U266增殖及凋亡作用的实验研究

Effect of SU11248 on proliferation and apoptosis of myeloma cell line U266

目的:探讨SU11248对骨髓瘤细胞U266的生物学效应的影响及其作用机制。方法:应用MTT法检测SU11248对U266细胞增殖能力的影响;用流式细胞技术检测细胞的DNA倍体及细胞周期变化、用凝胶电泳分析DNA片段化、用脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(TUNEL)检测SU11248对U266细胞凋亡的影响;以RT-PCR法检测3.0μg/mlSU11248作用U266细胞后c-myc、hTERT、Bcl-2、BaxmRNA表达变化。结果:SU11248可明显抑制U266细胞增殖(P<0.05),呈现剂量和时间依赖性,半数抑制浓度(IC50)约为3.0μg/ml。SU11248可将U266细胞阻滞于G0/G1期,并呈现剂量依赖性;能诱导U266细胞呈现典型的DNA梯状带;促进细胞原位凋亡。3.0μg/mlSU11248作用后,U266细胞c-myc、hTERT、Bcl-2mRNA表达水平呈时间依赖性降低,而BaxmRNA表达水平呈时间依赖性增强。结论:SU11248能抑制U266细胞增殖,诱导其凋亡,其作用机制可能与下调U266细胞Bcl-2、c-myc和hTERT的表达及上调Bax的表达有关。

Objective：To investigate the effect of SUl1248 proliferation and apoptosis of multiple myeloma cell line U266 in vitro and analyze its mechanisms. Methods：Effect of SUl1248 on proliferation of U266 ceils was detected by MTT assay.The ability of SUl1248 to induce apoptosis of U266 cells was examined by cell cycle analysis,TUNEL and DNA fragmentation. Expression of c-myc, hTERT, Bcl-2 and Bax mRNA in U266 cells was assessed by RT-PCR analysis.Results：The proliferation of U266 cells was inhibited by SUl1248 in dose- and time- dependent manners （P 〈 0.05）. The concentration of 50% growth inhibition （ICS0） of SU11248 for U266 was 3.0 μg/ml. SUl1248 induced G0/G1 phase of U266 cells, and induced U266 cell apoptosis as measured by detection of DNA fragmentation, flow cytometry and TUNEL. These data showed that SUl1248 induced apoptosis for U266 cells in dose- and time- dependent manners. After treating U266 cells with 3.0 μg/ml SU11248 at different time points, expressing level of c-myc, hTERT and Bcl-2 mRNA was reduced significantly and expressing level of Bax mRNA was increased significandy in a time-dependent manner（ P 〈 0.05）. Conclusion： SU11248 can inhibit the growth of U266 cells and induce the apoptosis of U266 cells. Its mechanism may be closely related with reduction the expression of c-myc, hTERT, Bcl-2 mRNA and enhancement the expression of Bax mRNA.