Bax和Fas在真菌多糖体外诱导HL-60细胞凋亡中的变化与作用

Changes and function of Bax and Fas in HL-60 cell apoptosis induced by two fungus polysaccharides

目的研究真菌Polyporus sp-M05多糖APS-1（一种多孔菌多糖SP．M05—1）和APS-2对白血病细胞HL-60增殖抑制及诱导凋亡作用，并探讨其分子机制。方法运用活细胞计数法检测多糖的增殖抑制作用并选择药物浓度；碘化啶（PI）染色后利用流式细胞术检测细胞凋亡作用；逆转录聚合酶链反应（RT—PCR）检测凋亡相关基因表达。结果多糖对HL-60细胞有增殖抑制作用，并随着浓度的增大和作用时间延长作用增强；APS-1作用48h后凋亡率达47．9％，APS-2作用细胞48h后凋亡率为26．8％，与阴性对照相比差异有统计学意义（P〈0．01）；RT-PCR检测到APS-1作用后细胞中Bax、Fas及半胱天冬酶（Caspase）-3基因上调，APS-2作用后细胞中Bax和Caspase-3基因上调，而Fas基因表达无变化。结论Polyporus sp．M05中提取的多糖APS-1和APS-2对白血病细胞HL-60具有增殖抑制和诱导凋亡作用，APS-1可同时通过线粒体途径和死亡受体途径诱导HL-60细胞凋亡，而APS-2通过线粒体途径诱导HL-60细胞凋亡。

Objective To detect the proliferation inhibition and apoptosis of HL-60 cell induced by APS-1 and APS-2 isolated from Polyporus sp. M05 and to investigate its mechanism. Methods The proliferation inhibition was detected by living cells count method,and chosed proper concentration. Flow cytometry with propidium iodide staining was used to detect cell apoptosis. Semi-quantitative RT-PCR was used to detect the expression of apoptosis related gene. Results AlaS-1 and APS-2 could significantly inhibit the proliferation of HL- 60 cells on a time and dose dependent manner. Apoptosis ratio increased to 47.9% and 26.8% after HL-60 cells were exposed to APS-1 and APS-2 respectively for 48 h, and the differences had statistical significance （P 〈0.01 ）. After being induced by APS-1, mRNA of Bax, Fas,Caspase-3 was upregulated. And after being induced by APS-2, mRNA of Bax, Caspase-3 was upregulated,while Fas mRNA did not change. Conclusion APS- 1 and APS-2 can inhibit the proliferation and induce apoptosis of HL-60 cells. Mechanism of HL-60 cell apoptosis induced by APS-1 is related to both mitochondrial pathway and Fas signaling pathway, while apoptosis induced by APS-2 is only related to mitochondrial pathway.