湖南师范大学学报（医学版）
湖南師範大學學報（醫學版）
호남사범대학학보（의학판）
JOURNAL OF HUNAN NORMAL UNIVERSITY(MEDICAL SCIENCE)
2013年
1期
10-14
,共5页
周蓓%肖立红%肖荞%任凯群%曹建国
週蓓%肖立紅%肖蕎%任凱群%曹建國
주배%초립홍%초교%임개군%조건국
肝细胞癌%肝癌干细胞样细胞%8-溴-7-甲氧基白杨素%自我更新
肝細胞癌%肝癌榦細胞樣細胞%8-溴-7-甲氧基白楊素%自我更新
간세포암%간암간세포양세포%8-추-7-갑양기백양소%자아경신
hepatocellular carcinoma%liver cancer stem cell like cell%8-bromo-7-methoxychrysin%self-re-newal
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。
目的：研究8-溴-7-甲氧基白楊素(BrMC)抑製源自人肝細胞癌 Huh-7細胞繫細胞腫瘤毬形成細胞即肝癌榦細胞樣細胞(LCSLCs)自我更新作用。方法：體外培養人肝癌Huh-7細胞繫細胞，榦細胞條件培養基懸浮培養法富集和擴增LCSLCs。 MTT比色法測定細胞活性；腫瘤毬形成法檢測自我更新能力；FITC-CD133標記FCM分析CD133錶達。結果：榦細胞條件培養基懸浮培養6天，Huh-7細胞形成非粘附、三維生長的肝癌毬體細胞即LCSLCs。與Huh-7細胞相比，BrMC抑製LCSLCs細胞活性作用更彊(P＜0.05)，併以濃度依賴性方式減少腫瘤毬數目(P＜0.05)，降低CD133蛋白錶達(P＜0.05)。結論：榦細胞條件培養基懸浮培養能分離和富集LC-SLCs；BrMC具有抑製LCSLCs細胞自我更新能力作用，其作用機製與降低榦細胞標記物CD133蛋白的錶達相關。
목적：연구8-추-7-갑양기백양소(BrMC)억제원자인간세포암 Huh-7세포계세포종류구형성세포즉간암간세포양세포(LCSLCs)자아경신작용。방법：체외배양인간암Huh-7세포계세포，간세포조건배양기현부배양법부집화확증LCSLCs。 MTT비색법측정세포활성；종류구형성법검측자아경신능력；FITC-CD133표기FCM분석CD133표체。결과：간세포조건배양기현부배양6천，Huh-7세포형성비점부、삼유생장적간암구체세포즉LCSLCs。여Huh-7세포상비，BrMC억제LCSLCs세포활성작용경강(P＜0.05)，병이농도의뢰성방식감소종류구수목(P＜0.05)，강저CD133단백표체(P＜0.05)。결론：간세포조건배양기현부배양능분리화부집LC-SLCs；BrMC구유억제LCSLCs세포자아경신능력작용，기작용궤제여강저간세포표기물CD133단백적표체상관。
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.
