【摘要】 目的 :确定组蛋白脱乙酰基酶抑制剂(HADCI)曲古霉素A(TSA)抑制人肝细胞癌MHCC97H源性球细胞组蛋白脱乙酰基酶-1(HADC1)表达以及自我更新作用。方法 :比色法测定HADC1活性;实时荧光定量PCR分析HADC1 mRNA表达;Western blot分析HADC1蛋白表达水平;球形成培养法检测肿瘤球形成能力。结果 :与单层贴壁生长细胞比较,MHCC97H源性球细胞高表达HDAC1,并具有更强自我更新能力。TSA下调MHCC97H源性球细胞HADC1表达,并显著抑制其自我更新能力。结论 :抑制HADC1表达参与曲古抑菌素A抑制MHCC97H源性球细胞自我更新作用。更多还原

【Abstract】 Objective To examine whether Trichostatin A, a histone deacetylase inhibitor, inhibits the expression of histone deacetylase-1(HADC1) and the capacity of self-renewal in spheres derived from hepatocellular cancer MHCC97 H cell line. Methods The relative activity of HADC1 was analyzed by colorimetric assay. The expression level of mHADC1 was analyzed by Quantitative Real-time PCR(qRT-PCR). The levels of HADC1 protein expressions were analyzed by Western blot. The capacity of Sphere-formation in vitro was evaluated by spheroid formation assay. Results The spheres exhibited elevated sphere formation capacity compared with the parental MHCC97 cells. TSA significantly suppressed the capacity of self-renewal in the spheres. The express levels of HADC1 were increased in the spheres, compared with parental cells. TSA effectively downregulated the expression of HADC1 at mRNA and protein levels in the spheres. Conclusion TSA inhibit self-renewing capacity of spheres from MHCC97 H cell line is associated with down-regulation of HADC1 expression.更多还原