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目的研究镰刀菌属真菌产生的倍半萜烯类化合物T-2毒素对体外培养急性早幼粒白血病细胞HL60生长抑制与凋亡诱导的作用。方法体外培养的HL60细胞经0、4、8、16和32μg/ml的T-2毒素处理48h,以MTT法检测T-2毒素对HL60细胞的生长抑制作用;吉姆萨染色观察细胞凋亡形态;流式细胞术(FCM)检测凋亡率;FCM及Western blot检测凋亡相关蛋白Bax、Bcl-2的表达情况。结果 MTT显示T-2毒素对HL60细胞的增殖具有抑制作用,并呈剂量依赖关系。形态学观察显示早幼粒白血病细胞出现明显凋亡形态特征。FCM定量检测表明,T-2毒素各处理组细胞凋亡率均高于对照组,并随T-2毒素浓度升高而增加。FCM和Western blot结果显示,T-2毒素处理后HL60细胞Bax蛋白表达上调,Bcl-2蛋白表达下降。结论T-2毒素可剂量依赖地抑制体外培养的HL60细胞增殖并诱导凋亡。
Objective To study the effect of sesquiterpene T-2 toxin induced by Fusarium fungi on the growth inhibition and apoptosis induction of acute promyelocytic leukemia cell line HL60 in vitro. Methods HL60 cells cultured in vitro were treated with T-2 toxin at 0, 4, 8, 16 and 32μg / ml for 48 hours. MTT assay was used to detect the inhibitory effect of T-2 to HL60 cells. FCM and FCM were used to detect the expression of Bax and Bcl-2. Results MTT showed that T-2 toxin could inhibit the proliferation of HL60 cells in a dose-dependent manner. Morphological observation showed that the promyelocytic leukemia cells showed obvious morphological features of apoptosis. FCM quantitative detection showed that T-2 toxin in each treatment group apoptosis rate was higher than the control group, and with the T-2 toxin concentration increased. FCM and Western blot showed that Bax protein expression was up-regulated and Bcl-2 protein expression was decreased in HL60 cells treated with T-2 toxin. Conclusion T-2 toxin can inhibit proliferation and induce apoptosis of HL60 cells cultured in vitro in a dose-dependent manner.