聚酯型儿茶素诱导HL-60细胞凋亡

The induction of apoptosis by theasinesin in HL-60 cells

目的 探讨聚酯型儿茶素的抗癌作用机制。方法 应用MTT法、电镜、琼脂糖凝胶电泳、流式细胞术、RT PCR和免疫细胞化学等方法研究细胞凋亡及其机制。结果 聚酯型儿茶素作用于HL 6 0细胞后 ,扫描和透射电镜观察到异染色质固缩、边集及凋亡小体形成 ;DNA琼脂糖凝胶电泳上可见典型的“阶梯状”条带 (DNALadder) ;AnnexinV FITC和PI双染后流式细胞术分析显示 ,在一定时间一定剂量范围内 ,聚酯型儿茶素诱导细胞凋亡的作用呈现浓度和时间依赖性增强的趋势 ,但是大剂量或长时间将主要导致细胞坏死 ;聚酯型儿茶素作用于HL 6 0细胞 2 4h后 ,细胞bcl 2基因及蛋白产物表达明显降低。结论 聚酯型儿茶素可诱导HL 6 0细胞调亡 。

AIM To investigate the mechanism of the anticancer effect of theasinesin. METHODS MTT assay, electronic microscopy technique, DNA gel electrophoresis, flow cytometric analysis with Annexin V Fluos staining,RT PCR and immunocytochemistry were performed for apoptosis and its mechanism. RESULTS After HL 60 cells were treated with theasinesin, HL 60 cells were characterized by condensation of the nuclear chromatin, margination against the nuclear envelope, and apoptotic body formation with scanning and transmission electronic microscopy. A typical DNA ladder was detected by agarose gel electrophoresis of DNA extracted from HL 60 cells. Flow cytometric analysis of HL 60 cells by Annexin V Fluos and propidium iodide double staining demonstrated that theasinesin induced apoptosis had dose effect and time effect relationships, but treatment with high dose or long time would induce cell necrosis principally. RT PCR and immunocytochemistry indicated that bcl 2 gene was reduced strongly in HL 60 cells. CONCLUSION\ Theasinesin induces apoptosis in HL 60 cells. The mechanisms for antitumor activity of theasinesin may involve in induction of apoptosis in cancer cells.