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目的:探讨中药臭灵丹中黄酮类化合物诱导人喉癌细胞Hep-2凋亡的机制。方法:MTT法检测分离自臭灵丹的黄酮类化合物3,5-二羟基-6,7,3’,4’-四甲氧基黄酮(HTMF)对2种正常细胞的毒性和对3种肿瘤细胞株的增殖抑制作用;采用流式细胞仪检测化合物对Hep-2细胞凋亡率的影响;Western blotting法检测凋亡蛋白caspase-3和caspase-9的变化。结果:HTMF显著抑制Hep-2细胞的增殖并呈浓度、时间双重依赖性关系,但对正常细胞Vero和EVC304的毒性较小,对A549和HepG2细胞抑制作用小。流式细胞仪检测结果显示HTMF对Hep-2细胞有促凋亡作用并呈明显的量效、时效关系。Western blotting结果显示HTMF可诱导Hep-2细胞中caspase-3和caspase-9蛋白的活化,并呈时间依赖性关系。结论:HTMF对人喉癌细胞Hep-2的生长有显著的抑制作用,其机制可能通过激活caspase-9进而活化caspase-3诱导Hep-2细胞凋亡。
OBJECTIVE: To investigate the mechanism of apoptosis of Hep-2 in human laryngeal carcinoma cells induced by flavonoids from traditional Chinese medicine Silanol. Methods: The toxicity of 3, 5-dihydroxy-6, 7, 3 ’, 4’-tetramethoxyflavone (HTMF) The effect of compound on the apoptosis rate of Hep-2 cells was detected by flow cytometry. The changes of caspase-3 and caspase-9 were detected by Western blotting. Results: HTMF significantly inhibited the proliferation of Hep-2 cells in a dose-dependent and time-dependent manner. However, HTMF had less cytotoxic effects on Vero and EVC304 cells and less inhibitory effect on A549 and HepG2 cells. The results of flow cytometry showed that HTMF could promote the apoptosis of Hep-2 cells with obvious dose-effect and time-effect relationship. Western blotting showed that HTMF could induce the activation of caspase-3 and caspase-9 in Hep-2 cells in a time-dependent manner. CONCLUSION: HTMF can significantly inhibit the growth of human laryngeal carcinoma Hep-2 cells. The mechanism may be through activation of caspase-9 and activation of caspase-3 to induce Hep-2 cell apoptosis.