ADAM10抑制剂GI254023X对H929细胞增殖与凋亡的作用及其机制研究

Effect of ADAMIO Inhibitor GI254023X on Proliferation and Apoptosis of Multiple Myeloma H929 Cells and Its Possible Mechanisms

目的:探讨解聚素金属基质蛋白酶10(ADAM10)抑制剂GI254023X对多发性骨髓瘤H929细胞增殖与凋亡的影响及其作用机制。方法:选择不同浓度GI254023X处理H929细胞,CCK-8法检测并绘制增殖抑制曲线,Annexin V/7-AAD双染流式细胞术检测细胞存活与凋亡,Western blot检测切割后的Notch1蛋白(Cleaved Notch1),定量PCR检测Notch1靶基因Hes-1的转录变化。结果:GI254023X能明显抑制H929细胞增殖,随着药物浓度的增大及时间的延长,细胞的增殖能力逐渐下降;GI254023X能够诱导H929细胞凋亡;Western blot检测结果显示,Cleaved Notch1在GI254023X作用后水平下降;GI254023X能够使Hes-1基因的表达减低。结论:GI254023X能抑制H929细胞增殖并诱导其凋亡,其机制可能与抑制Notch1活化有关。

Objective:To investigate the effect of ADAM 10 inhibitor GI254023 X on the proliferation and apoptosis of multiple myeloma H929 cell line and its mechanisms.Methods:H929 cells were treated with different concentrations of GI254023 X,the proliferation-inhibitive curve was assayed and plotted by CCK-8 method,the cell viability and apoptosis were detected by flow cytometry with Annexin V/7-AAD double staining.The cleavage of Notch1 protein(cleaved notch1) was determined by Western blot.The transcripts of Notch1 target gene Hes-1 were detected by realtime PCR.Results:The GI254023 X inhibited the proliferation of H929 cells in the time-and dose-dependent manners.As compared with the control group,the apoptosis of cells increased along with enhancement of GI254023 X concentration;The expression of cleaved Notch1 was down-regulated after the treatment with GI254023 X.The levels of Hes-1 mRNA transcripts in H929 cells was reduced in GI254023 X treated group.Conclusion:GI254023X can remarkably inhibit the proliferation and induce the apoptosis of H929 cells.Its mechanism may be associated with inbihition of Notchl activation.