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Dihydroartemisinin induced apoptosis of peripheral T lymphocytoma Hut-78 cells and its possible mechanism
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Abstract:
Objective:To investigate dihydroartemisinin (DHA)-induced apoptosis of peripheral T lymphocytoma Hut-78 cell line and its possible mechanism. Methods: The effect of DHA 1－30 μg/ml on proliferation of Hut-78 cells was measured by CCK-8 assay. The morphological changes of Hut-78 cell's nuclei induced by DHA were observed by Hochest 33258 staining and confocal microscopy. Flow cytometry was used to examine the apoptosis of Hut-78 cells induced by DHA. After pretreating the Hut-78 cells with 10 mmol/L of reactive oxygen species (ROS) scavenger N-acetyl cysteine C (NAC) , a role of ROS in changes of mitochondrial membrane potential induced by DHA was evaluated. Effect of ROS on release of cytochondrial C induced by DHA during apoptosis of Hut-78 cells was examined with an immunoblotting assay.Results:DHA inhibited proliferation of the Hut-78 cells in a dose-dependent manner, induced nucleus pyknosis and formed apoptotic bodies in the cells. Apoptosis rates of the Hut-78 cells iuduced with 5, 10 and 20 μg/ml of DHA were (25.1±2.8)%, (43,6±3.1)% and (68.9±2.6)% respectively, which were significantly different from that of the control group (all P<0.01 ). Furthermore, treatment with 20 μg/ml of DHA droped mitochondrial membrane potential of the Hut-78 cells by (59.4±2.6)%, and pretreating with ROS scavenger NAC droped the mitochondrial membrane potential by (38.4±2.1)%. DHA can induce release of cytochrome C in mitochondrion, which can be significantly inhibited by pretreatment with NAC. Statistic results further demonstrated this point. Conclutions: DHA can effectively inhibit proliferation of the peripheral T lymphocytoma Hut-78 cells and induce their apoptosis. Its action mechanism may be related to ability of HAD to promote release of ROS-dependent cytochrome C.
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Project Supported:
Project supported by the Foundation from the Bureau of Education of Henan Province（No. 2006320039）
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