冬凌草甲素诱导肝癌细胞凋亡作用及其机制  

Oridonin-induced apoptosis of hepatocellular carcinoma cells and mechanism

作　　者：郭文治[1,2] 王冬雨[3] 李功权[1,2] 张嘉凯[1,2] 温培豪[1,2] 潘洁[1,2] 史晓奕[1] 张水军[1,2] 

机构地区：[1]郑州大学第一附属医院肝胆外科,450003 [2]河南省消化器官移植重点实验室 [3]郑州大学第一附属医院感染科,450003

出　　处：《中华实验外科杂志》2015年第9期2157-2159,共3页Chinese Journal of Experimental Surgery

基　　金：郑州市科技局基金资助项目（131PCXTD617）;河南省科技攻关计划项目（122102310215）;河南省高等学校重点科研项目计划（15A320038）

摘　　要：目的 探讨冬凌草甲素（Oridonin）诱导肝癌细胞凋亡作用及其分子机制.方法 体外培养肝癌细胞株BEL-7402、SK-Hep1,采用不同浓度（0～ 10 μmol/L）的Oridonin处理细胞,WST-1方法检测细胞的增殖率,锥虫蓝拒染法检测死细胞率,流式细胞学方法检测细胞的凋亡率,Western blot检测半胱氨酸天门冬氨酸蛋白酶-3（Caspase-3）和聚腺苷二磷酸核糖聚合酶（PARP）蛋白,以及抗凋亡B细胞淋巴瘤/白血病-2（bcl-2）家族蛋白bcl-2、bcl-xl、髓样细胞白血病-1（Mcl-1）表达.结果 WST-1检测结果显示,Oridonin作用于肝癌细胞株SK-Hep-1、BEL-7402细胞24h后的半数抑制剂量（IC50）分别是1.58、1.68 μmol/L;锥虫蓝拒染法检测显示,5μmol/L的Oridonin分别处理SK-Hep-1和BEL7402细胞48 h,2株肿瘤细胞的死亡率分别为96.54％和86.54％,相对于对照组差异均有统计学意义（P＜0.01）.流式细胞仪检测显示,5 μmol/L的Oridonin分别处理SK-Hep-1和BEL7402细胞24h,2株细胞处理组细胞凋亡率分别为65％和44％;Western blot检测显示,Oridonin处理肝癌细胞24 h后,在两株肝癌细胞中均可检测到Caspase-3活化、PARP裂解,Mcl-1表达下调.结论 Oridonin对肝癌细胞有强效的诱导凋亡作用,其分子机制可能与下调Mcl-1后凋亡信号通路活化有关.Objective To investigate activity and the underlying mechanism of oridonin-mediated anticancer activity in hepatocellular carcinoma （HCC） cells.Methods HCC cell lines BEL-7402 and SK-Hepl were treated with different concentrations （0-10 μmol/L） of oridonin,cell proliferation was examined by WST-1 assay,cell death was examined by trypan blue staining,apoptosis was examined by flow cytometry,and the expression of Caspase-3,cleaved poly adenosine diphosphate-ribose polymerase （PARP） and anti-apoptotic bcl-2 family members was analyzed by Western blotting.Results WST-1 assays showed that oridonin dose-dependently inhibited cell proliferation in both HCC cell lines,with 50％ inhibitory dose （IC50） values of 1.58 μmol/L and 1.68 μmol/L,respectively,in SK-Hep-1,BEL-7402 cell lines.Cell death assays showed treatment with oridonin at 5 μmol/L for 48 h induced 96.54％ and 86.54％ cell death,respectively,in SK-Hep-1 and BEL7402 cell lines.Flow cytometry showed that oridonin increased apoptosis rate by 65％ and 44％,respectively,in SK-Hep-1 and BEL7402 cell lines.Western blotting showed that treatment with oridonin at 5 μmol/L for 24 h induced activation of Caspase-3 and cleavage of PARP in both cell lines.Moreover,treatment with oridonin at 2.5,5 and 10 μmol/L for 24 h markedly inhibited myeloid cell leukemia-1 （Mcl-1） expression in both cell lines in a dose-dependent manner.Conclusion Oridonin potently inhibited proliferation of HCC cells.The strong anti-HCC activity may be associated with the degradation of antiapoptotic protein Mcl-1,and the activation of apoptotic signaling pathway.

关 键 词：冬凌草甲素 癌 肝细胞 脱噬作用 髓样细胞白血病.1基因 

分 类 号：R282.71[医药卫生—中药学]