木犀草素对MHCC97H源性肝癌干样细胞HADC1活性和自我更新的影响

Effects of luteolin on HADC1 activity and self-renewal of liver cancer stem-like cells in MHCC97H cell line

目的:本研究旨在阐明木犀草素(LUT)是否通过抑制组蛋白脱乙酰基酶-1(HADC1)活性和表达从而抑制肝细胞癌MHCC97H细胞源性干样细胞(LCSLCs)自我更新能力。方法:LUT(10μM)和HADC1抑制剂曲古抑菌素A(TSA,10μM)单独或联合处理MHCC97H源性LCSLCs。采用比色法测定HADC1活性;实时荧光定量PCR分析HADC1 mRNA表达;蛋白质印记分析HADC1蛋白表达水平;球形成培养法检测肿瘤球形成能力。结果:LUT抑制MHCC97H源性LCSLCs HADC1活性,下调HADC1表达,并抑制其自我更新。与LUT或TSA单独处理组相比,LUT与TSA联合处理协作性下调MHCC97H源性LCSLCs HADC1表达和活性;并显著抑制MHCC97H源性LCSLC球形成。结论:LUT部分地通过下调HADC1活性和表达抑制MHCC97H源性LCSLC自我更新。

Objective This study aims to clarify whether Luteolin(LUT)inhibits self-renewal ability of MHCC97H cells-derived liver cancer stem cell-like cells(liver cancer stem-like cells,LCSLC)by inhibiting histone deacetylase-1(HADC1)activity and expression.Methods MHCC97H-derived LCSLC was treated with LUT(10μM)and HADC1 inhibitor trichostatin A(TSA,10μM)alone or in combination.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 LUT down-regulates HADC1 expression in MHCC97H-derived LCSLC and inhibits its self-renewal ability.Compared with the application of LUT or TSA alone,the combination of LUT and TSA has a stronger cooperative effect on the down-regulation of HADC1 expression in MHCC97H-derived spheroid cells.Compared with the application of LUT or TSA alone,the combination of LUT and TSA significantly inhibits its self-renewal ability.Conclusion LUT inhibits the self-renewal of MHCC97H-derived LCSLC partly by down-regulating the activity and expression of HADC1.