中药臭灵丹中黄酮类化合物的体外抗肿瘤活性研究

Research on antiproliferative effect of flavones isolated from Laggera pterodonta

目的:探讨中药臭灵丹中黄酮类化合物的体外抗肿瘤活性。方法:采用MTT法,检测分离自臭灵丹的6个黄酮类化合物对2种正常细胞的毒性和对2种肿瘤细胞的增殖抑制作用,并初步分析它们的构效关系;运用流式细胞仪,检测活性化合物对细胞凋亡率及细胞周期的影响。结果:6个黄酮化合物中,5,7,3,′4′-四甲氧基-3-羟基黄酮与金腰素乙显著抑制A549和HeLa 2种肿瘤细胞的增殖并呈浓度依赖性关系,且对正常细胞Vero和EVC304的毒性较小;这2个黄酮结构中均有邻位酚甲氧基。流式结果显示金腰素乙对HeLa细胞的G2/M期起阻滞作用并呈浓度依赖性关系。5,7,3,′4′-四甲氧基-3-羟基黄酮与金腰素乙对A549和HeLa细胞都有促凋亡作用并呈明显的量效关系。结论:5,7,3,′4′-四甲氧基-3-羟基黄酮与金腰素乙对肿瘤细胞的生长有显著的抑制增殖作用且细胞毒性小,并通过调节细胞周期及诱导凋亡发挥抗肿瘤作用。

Objective： To research the cytotoxicity and in vitro antiproliferative effect of the six flavone compounds extracted from Laggera pterodonta.Method： The cytotoxicity on the normal cells and antiproliferative effect on tumor cells were tested by MTT assay,and then the preliminary structure-activity relationship was analysed.The phase distribution of the cell cycle and apoptosis rate were analyzed by flow cytometry.Result： The results of MTT assay showed 5,7,3′,4′-tetramethoxy-3-hydroxyflavone and chrysosplenetin B inhibited growth of A549 and Hela cells significantly with a dose dependent mode,while exhibited low cytotoxicity to the two normal cells,Vero and EVC304.Both compounds contain ortho-phenolic methoxyl moietys in their structures.Flow cytometry analysis revealed that Hela cells treated with increasing quantities of chrysosplenetin B increased the percentage of cells in the G2/M phase,and Hela and A549 cells treated with increasing quantites of the 5,7,3′,4′-tetramethoxy-3-hydroxyflavone and chrysosplenetin B increased the apoptosis rates.Conclusion： The 5,7,3′,4′-tetramethoxy-3-hydroxyflavone and chrysosplenetin B extracted from L.pterodonta showed high antiproliferative effect on cancer cells with low cytotoxicity on normal cells,and took the effects on A549 and Hela cells through the hold-up of the G2/M phase of the cell cycle and induction of the apoptosis.