原花青素诱导HL-60细胞分化及其机制的研究  

Differentiation of Human Promyelocytic Leukemia HL-60 Cells Induced by Proanthocyanidin and Its Mechanism

作　　者：谢朝阳[1] 吴斌华[1] 杨志刚[2] 陈小芳[1] 陈秋生[1] 

机构地区：[1]广东医学院检验医学研究所,广东湛江524023 [2]广东医学院附属医院血液科,广东湛江524001

出　　处：《中国实验血液学杂志》2013年第4期920-925,共6页Journal of Experimental Hematology

基　　金：东莞市科技计划资助项目(编号2008106101065);广东省中医药局课题资助项目(编号2007129)

摘　　要：本研究旨在探讨原花青素(proanthocyanidin,PAC)诱导人急性早幼粒细胞白血病细胞株HL-60细胞分化的可能机制。用PAC 20 mg/L处理HL-60细胞24 h,CCK-8法检测细胞生长状况,分析PAC对HL-60细胞的增殖影响;应用光学显微镜观察细胞形态变化,流式细胞术分析细胞周期变化和测定细胞分化抗原CD14、CD11b表达;Western blot检测P21、Cyclin D1和CDK4表达变化。结果表明,PAC 20 mg/L作用HL-60细胞24 h,细胞抑制率明显增高(73.2±1.5)%(P<0.01),髓系细胞分化抗原CD14表达明显增高(P<0.01),CD11b表达稍增高,细胞周期中G0/G1期细胞百分率明显增高(P<0.05),S期细胞明显减少(P<0.01);PAC 20 mg/L与HL-60细胞共培养4 h,部分细胞向成熟阶段细胞分化;PAC 20 mg/L处理HL-60细胞12、24和48 h,P21蛋白表达增加,Cyclin D1和CDK4蛋白表达降低。结论:PAC能抑制HL-60细胞增殖并诱导其分化,细胞周期阻滞于G0/G1,其机制可能与P21蛋白表达上调和Cyclin D1及CDK4蛋白表达下调有关。This study was purposed to investigate the proliferation,differentiation and apoptosis of human promyelocytic leukemia HL-60 cells induced by proanthocyanidin（PAC）.HL-60 cells were incubated with 20 mg/L PAC for 24 h,the cell growth was evaluated by CCK-8 assay.the effect of PAC on HL-60 cells was evaluated and the cells morphology was observed by optical microscopy.Expression of CD14 and CD11b,and cell cycle were analyzed by flow cytometry.The results showed that the growth of HL-60 cells was inhibited after treatment with PAC of different concentration in a dose-dependent manner（P〈0.05）.20 mg/L PAC displayed significant effect on HL-60 cells with inhibition ratio（72.3±1.8）% for 24 h.Microscopy displayed that some cells differentiated to relative mature cells after treating for 48 h.Expression of CD14 increased and the expression of CD11b increased a little after treating with 20 mg/L PAC for 24 h,the ratio of cells in G0/G1 phase increased,but the ratio of cells in S phase discreased.The mRNA and protein expression of P21 gene increased,but the protein expression of CDK4 and Cyclin D1 decreased.It is concluded that PAC may inhibit the proliferation of HL-60 cells in vitro,induces the differentiation of HL-60 cells,and arrests the cells in G0/G1 phase.The possible mechanism may be related to up-regulation of P21 gene expression and down-regulation of the protein expression of CDK4 and Cyclin D1.

关 键 词：原花青素 白血病 HL-60细胞 诱导分化 

分 类 号：R733.7[医药卫生—肿瘤]