8-溴-7-甲氧基白杨素对Huh-7细胞系肝癌干细胞样细胞自我更新的影响

Effects of 8-bromo-7-methoxychrysin on self-renewal of liver cancer stem cell like cells from human hepatocellular carcinoma Huh-7 cell line

目的:研究8-溴-7-甲氧基白杨素(BrMC)抑制源自人肝细胞癌Huh-7细胞系细胞肿瘤球形成细胞即肝癌干细胞样细胞(LCSLCs)自我更新作用。方法:体外培养人肝癌Huh-7细胞系细胞,干细胞条件培养基悬浮培养法富集和扩增LCSLCs。MTT比色法测定细胞活性;肿瘤球形成法检测自我更新能力;FITC-CD133标记FCM分析CD133表达。结果:干细胞条件培养基悬浮培养6天,Huh-7细胞形成非粘附、三维生长的肝癌球体细胞即LCSLCs。与Huh-7细胞相比,BrMC抑制LCSLCs细胞活性作用更强(P<0.05),并以浓度依赖性方式减少肿瘤球数目(P<0.05),降低CD133蛋白表达(P<0.05)。结论:干细胞条件培养基悬浮培养能分离和富集LC-SLCs;BrMC具有抑制LCSLCs细胞自我更新能力作用,其作用机制与降低干细胞标记物CD133蛋白的表达相关。

Objective The aim of this study was to examine the effect of 8-bromo-7-methoxychrysin (BrMC) on self-renewal of the sphere forming cells derived from human hepatocellular carcinoma (HCC) cell line Huh-7 cultured in vitro, namely liver cancer stem cell like cells (LCSLCs). Methods The stem cell conditioned culture system was used to isolate and enrich LCSLCs. After treated with various concentrations of BrMC, the cell viability of LCSLCs was examined by MTT assay, using tumorsphere formation assay to determine the self-renewal and FCM analysis with FITC-CD133 to assess the expression levels of CD133. Results Sphere-forming cells, namely LCSLCs were developed after Huh-7 cells were cultured in stem cell conditioned culture system for 6 days, the spheroids are character by nonadherent, three-dimensional. Compared with the parent Huh-7 cells, BrMC has more stronger effects of inhibiting cell viability in LCSLCs (P<0.05), decreased the amount of spheroids (P<0.05) in concentration-dependent manner and downregulated the level of CD133 protein expression. Conclu-sion Stem cell conditioned culture system could effectively isolated and enriched LCSLCs derived from HCC line Huh-7; BrMC possesses the inhibitory effect on self-renewal and those are associated with down-regulation of stem cell marker CD133 expression in LCSLCs.