组蛋白去乙酰化酶抑制剂LBH589体外诱导人多发性骨髓瘤细胞株U266凋亡及其机制

目的研究新一代组蛋白去乙酰化酶（HDAC）抑制剂LBH589单药或联合中药禹州漏芦含药血清对人多发性骨髓瘤（MM）细胞株U266诱导凋亡作用及其机制。方法利用免疫印迹技术（Westernblot）分析不同浓度LBH589作用HDAC6特异底物α-微管蛋白乙酰化水平；采用免疫沉淀（IP）法检测不同浓度LBH589作用后热休克蛋白90（HSP90）与其客户蛋白的亲和力。结果Westernblot分析显示不同浓度LBH589（0、20、50nmol／L）单药及50nmol／L与禹州漏芦含药血清（1g／m1）联合均能够抑制U266细胞增殖，随着药物浓度的增加和作用时间的延长，仪。微管蛋白乙酰化水平、HSP90乙酰化的程度逐渐上调，变化呈剂量依赖性（P〈0．05），且LBH589与禹州漏芦含药血清联合组抑制作用较单药组明显（均P〈0．05）。结论LBH589能够抑制MM细胞增殖，阻滞细胞周期，诱导人MM细胞株U266凋亡。

Histone deacetylase inhibitor LBH589 induces apoptosis of multiple myeloma cell line U266 in vitro and its mechanism

Objective To study the effect of new generation histone deacetylase inhibitor LBH589 single-drug or combined with the mouse serum which contains the radix echinopsis on multiple myeloma (MM) cell line U266 and their mechanism.Methods The acetylation level of α-tublin which was a substrate of HDAC6 was detected by Western blot.The chemical force between Heat shock protein 90 (HSP90) and its client proteins was detected by immune precipitation (IP).Results The different concentrations of LBH589 single drug (0,20,50 nmol/L),and 50 nmol/L combined with the mouse serum which contained the radix echinopsis (1 g/ml) were able to inhibit the proliferation of U266 cell.With the increase of drug concentration and the extension of time,the acetylation levels of α-tublin and HSP90 increased gradually (at 24 or 48 hours) in a dose dependent (P ＜ 0.05).The inhibition of LBH589 combined with the mouse serum was stronger than that of LBH589 single drug (P ＜ 0.05).Conclusion LBH589 could inhibit the growth of MM cells and their cell cycles,and induce the apoptosis of MM cell line U266.