茶多酚对HL-60细胞株体外增殖及细胞周期的影响

目的 探讨茶多酚对人急性早幼粒白血病细胞株HL-60细胞增殖和细胞周期的影响.方法 采用四甲基偶氮唑盐微量酶反应比色法(MTT比色法)观察茶多酚对体外培养的HL-60细胞增殖活性的影响.采用HE染色、荧光染色观察用茶多酚后细胞形态变化.用流式细胞仪(FACS)检测细胞凋亡率及细胞周期.结果 (1)MTT比色法检测显示茶多酚能抑制HL-60细胞增殖,在一定范围内呈剂量和时间依赖性(P＜0.05).当茶多酚浓度达到400和800 mg/L时,48 h抑制率分别为(58.90±1.19)%和(72.57±0.70)%.(2)流式细胞仪分析,茶多酚处理组出现一特征性的亚二倍体凋亡峰.其凋亡率呈时间、剂量依赖性.(3)流式细胞术发现茶多酚可使HL-60细胞阻滞于S期,其阻滞细胞的数量与药物浓度呈正相剂量关系,以作用24 h对细胞周期的阻滞作用最强.结论 茶多酚能有效抑制HL-60细胞增殖,在一定范围内具有时间和剂量依赖性.茶多酚可体外诱导HL-60细胞凋亡,并使细胞周期阻滞于S期.

Effects of tea polyphenols on proliferation and cell cycle of HL-60 cell line in vitro

Objective To investigate the in vitro effects of tea polyphenols (TP) on proliferation and cell cycle of human acute promyelocytic leukemia cell line HL-60. Methods The in vitro effects of TP on proliferation of HL-60 cells were analyzed by MTT colorimetric assay. The morphologic changes of HL-60 cells after TP treatment were observed by HE staining and immunofluorescence staining, and flow cytometry was used to examine the cell apoptosis rate and cell cycle. Results ①The result of MTT colormetric assay revealed that TP inhibited the proliferation of HL-60 cells in dose- and time-dependent manner within a certain range of dosage (P＜0.05). With 400 mg/L or 800 mg/L TP, the 48 h inhibition rate of the HL-60 cells was (58.90± 1.19) ％ and (72.57±0.70) ％, respectively. ②By using flow cytometry, a distinctive hypodiploid peak was observed in HL-60 cells cultured with TP, and the apoptosis rate was dose- and time-dependent.(3) HL-60 cells were arrested at S phase and the number of arrested cells was positively correlated with drug dose. The strongest blockage effect of TP was observed at 24 h. Conclusions Within a certain range of dosage, TP can inhibit the proliferation of HL-60 cells in dose- and time-dependent manner. TP can induce HL-60 cells apoptosis and block the cell cycle of HL-60 cells at S phase in vitro.