海兔中新环氧甾醇的体外抗肿瘤活性研究

Studies on in vitro antitumor activities of a new sterol HEO from sea hare

目的:研究一种海兔中新环氧甾醇(HEO)的体外抗肿瘤活性。方珐:以体外培养的人早幼粒细胞白血病细胞株HL-60作为研究对象,采用3H-TdR掺入实验、Annexin V荧光染色检测、流式细胞术、透射电镜等观察HEO对细胞周期和诱导凋亡的作用,以考察HEO的抑瘤活性。结果:HEO对HL-60作用24 h的半数抑制浓度IC50为1.4 mg／L,在0.8 mg／L时已明显抑制HL-60细胞的对数生长;3H-TdR掺入实验表明,随着HEO作用时间的延长,对HL-60细胞的增殖抑制作用明显增强。Annexin V荧光染色表明HEO在作用24 h后,大量HL-60细胞开始出现凋亡。透射电镜显示肿瘤细胞染色质呈现凋亡表现。细胞周期分析显示G2+M期细胞增多。结论:HEO明显抑制HL-60生长,并通过诱导凋亡杀伤肿瘤细胞,抗肿瘤活性强;对细胞周期的影响提示其抑瘤机制可能为M期阻滞。

Objective:To study the in vitro antitumor activities of a new sterol HEO from sea hare. Methods : With 3H-TdR incorporation experiment, Annexin V fluorescence dye, flow cytometry, transmission electron microscope,human myel-ogenous leukemia cells (HL-60) cultured in vitro were tested to observe the effect of HEO on cell cycle and in inducing apop-tosis. Results:The IC50 of HEO to HL-60 in 24 h was 1. 4 mg/L. HEO obviously inhibited the logarithm growth of HL-60 at the concentration of 0. 8 mg/L. The experiment of 3H-TdR incorporation showed that the inhibitory effect on proliferation of HL-60 cells was apparently enhanced as the time of HEO action prolonged. Annexin V fluorescence dye showed that after 24 h HEO action,a large number of HL-60 cells became apoptotic. Transmission electron microscopy revealed that the chromatin of tumor cells presented with apoptosis features. The analysis of cell cycle showed that the number of G2 + M cells were increased. Conclusion :HEO can inhibit the growth of HL-60 and kill the tumor cells by inducing apoptosis. The influence on the cell cycle indicates that antitumor activities of HEO may result from M phase block.