岩藻黄质对人红白血病HEL细胞的凋亡作用及其机制

Mechanism and Apoptosis Effect of Fucoxanthin in Human Erythroleukemia Leukemia Cells

为探讨岩藻黄质对人红白血病HEL细胞株增殖抑制作用及其诱导凋亡机制,以岩藻黄质处理人红白血病细胞(HEL细胞),采用四甲基偶氮唑蓝(MTT)法检测细胞增殖,流式细胞仪检测细胞凋亡率,分析细胞周期,测定细胞线粒体膜电位,并应用实时荧光定量PCR(RT-qPCR)和Western blot法检测凋亡相关基因及蛋白表达的变化。结果表明,岩藻黄质呈剂量依赖性抑制HEL细胞增殖(P<0.01)。流式细胞术检测显示,岩藻黄质作用24 h后,HEL细胞早期和晚期凋亡的比率极显著增高(P<0.01),G0/G1细胞比例增多,S期和G2/M期细胞比例减少,线粒体膜电位降低;岩藻黄质通过上调促凋亡基因和下调抑凋亡基因的表达引起细胞的凋亡,Bcl-xL蛋白的表达变化不显著(P>0.05),Bcl-2蛋白的表达下调,Caspase-3和Bax蛋白的表达极显著上调(P<0.01)。由此可见,岩藻黄质能诱导HEL细胞凋亡,这为新型抗白血病功能食品的开发及白血病的治疗提供了理论依据。

This study was aimed to investigate the inhibitory effect of fucoxanthin on the proliferation of human erythroleukemia(HEL) cells and the mechanism of inducing apoptosis. The proliferation of HEL cell treated with different concentrations of fucoxanthin was assessed by methyl thiazolyl tetrazolium(MTT) assay, and the cell cycle, mitochondrial membrane potential and apoptosis changes were analyzed by flow cytometry. The expression of apoptosis-related genes and proteins were determined by the real-time quantitative PCR(RT-qPCR) and Western blot. The results showed that fucoxanthin decreased the proliferation of HEL cell in a concentration manner. Flow cytometry showed that fucoxanthin increased the percentage of HEL cells in the G0/G1 phase, reduced the percentage of HEL cells in the G2/M and S phase, promoted cell apoptosis and decreased mitochondrial membrane potential after treated with fucoxanthin for 24 h. It also induced apoptosis by up-regulating expression of pro-apoptotic genes and down-regulating the anti-apoptotic genes. The expression of Bcl-xL protein was no significant change. In addition, Caspase-3 and Bax protein levels were significantly increased, but the expression of Bcl-2 protein was decreased. The results indicated that fucoxanthin had a significantly inhibitory effect on the HEL cells in vitro, which provided a theoretical basis for the development of new anti-HEL functional foods.