Introduction

Dihydroartemisinin (DHA), a semi-synthetic derivative of artemisinin, has recently shown to induce apoptosis in many types of cancer cells. In this study, we aimed to determine the effects of DHA on apoptosis in human chronic lymphocytic leukemia (CLL) cell lines.

Methods

The cells were treated separately and combined by DHA and Fludurabine (FLU) during 24, 48 and 72 hours. The cell viabilities determined by XTT method. Following separate and combined treatment of IC50 concentrations of DHA and FLU to the cells during 24 hours, the cells were analyzed by flow cytometry to determine the effects on apopotis staining with AnnexinV FITC and PI. mRNA and protein expression levels of TCTP, Mcl-1, Bcl-2, Bax and Caspase-3 were analyzed to find out the molecular mechanisms of apoptosis by using quantitative real-time PCR and flow cytometric methods.

Results

Treatment with DHA alone or in combination with FLU induced apoptosis in a dose dependent manner in CLL cells. DHA alone was more effective than FLU alone or combined treatment with DHA and FLU. Our results suggest that Bcl-2 protein family member Bax was active in the apoptotic response of CLL cells after DHA treatment. Moreover, the apoptotic response induced by DHA was independent from the p53 mutation status of the CLL cells.

Conclusion

DHA might be a potential anti-cancer therapeutic for CLL.

中文翻译：

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双氢青蒿素对慢性淋巴细胞白血病细胞系凋亡影响的免疫分子评价

介绍

双氢青蒿素 (DHA) 是青蒿素的一种半合成衍生物，最近已显示可诱导多种癌细胞凋亡。在这项研究中，我们旨在确定 DHA 对人类慢性淋巴细胞白血病 (CLL) 细胞系细胞凋亡的影响。

方法

在 24、48 和 72 小时内分别处理细胞并通过 DHA 和氟杜拉滨 (FLU) 合并。XTT法测定细胞活力。在 24 小时内分别和联合处理 IC50 浓度的 DHA 和 FLU 对细胞后，通过流式细胞术分析细胞以确定用 AnnexinV FITC 和 PI 对细胞凋亡的影响。采用实时定量PCR和流式细胞术分析TCTP、Mcl-1、Bcl-2、Bax和Caspase-3的mRNA和蛋白表达水平，探讨细胞凋亡的分子机制。

结果

在 CLL 细胞中，单独使用 DHA 或与 FLU 联合治疗以剂量依赖性方式诱导细胞凋亡。单独使用 DHA 比单独使用 FLU 或与 DHA 和 FLU 联合治疗更有效。我们的结果表明 Bcl-2 蛋白家族成员 Bax 在 DHA 处理后的 CLL 细胞凋亡反应中是活跃的。此外，DHA 诱导的细胞凋亡反应与 CLL 细胞的 p53 突变状态无关。

结论

DHA 可能是 CLL 的潜在抗癌治疗剂。