Mechanism Investigation of Growth Inhibition and Apoptosis caused by Bruceine D in K562 Cells

• 作 者: Min-ting Lin Si-li Tang Hu-biao Chen Jian-ye Zhang
• 机 构: School of Pharmaceutical Sciences,Guangzhou Medical University,Guangdong 510182,China
• 出 处: 2014医学科学前沿暨第三届个体化治疗与抗肿瘤药物研究新趋向研讨会
• 发表日期: 2014年5期
• 关键词: Bruceine D Apoptosis mitochondrial pathway AKT ERK Phosphorylation

　　Objective: To observe the changes of mitochondrial membrane potential (MMP),cytochrome c (Cyto-c), Caspase-3, 9 activity, cleavage of PARP and phosphorylation level of AKT and ERK, investigate the mechanism of growth inhibition and apoptosis included in Bruceine D-treatment K562 cells.Methods: K562 cells were treated by different concentration of Bruceine D (3.0, 6.0,12.0 μM) for 24h.MTT assay was used to evaluate the cell growth inhibition of Bruceine D in vitro; Flow cytometry were performed to analyze MMP; Western Blot analysis was applied to detect expression of Cyto-c, Caspases-3,-9, PARP and other protein in K562 cells.Results: IC50 value of Bruceine D against K562 cells was 6.37±0.39 μM.The percentages of MMP after the treatment of 3.0,6.0,12.0 μM Bruceine D for 24 h were 79.84 ± 4.46%, 59.74 ± 7.48%, 40.66 ± 4.37% (P<0.05) respectively.The release of Cyto-c, activity of Caspase-3, 9 and cleavage of PARP increased compared with the control groups in the Bruceine D induced K562 cell.Moreover, Bruceine D could decrease phosphorylation level of AKT and ERK.Conclusions: The collapse of MMP, the increased Cyto-c, the up-regulation of Caspase-3,-9 activity and the augmented cleavage of PARP emerged after K562 cells treated by Bruceine D.The apoptosis of K562 cells induced by Bruceine D might be related to the mitochondrial pathway of apoptosis.Reduction of AKT and ERK phosphorylation level might be mechanism of growth inhibition of K562 cells mediated by Bruceine D.