[关键词]
[摘要]
目的观察金莲花中提取的牡荆苷体外对人食管癌EC-109细胞生长及诱导EC-109细胞凋亡的作用,探讨p53及bcl-2蛋白表达在牡荆苷抗肿瘤细胞机制中的作用。方法用不同浓度牡荆苷作用于对数生长期的EC-109细胞,通过CCK-8法检测其对EC-109细胞体外生长、增殖的抑制作用|Hoechst33258荧光染色观察细胞形态的变化|琼脂糖凝胶电泳检测DNA条带|AnnexinV-FITC/PI双标法流式细胞术观察牡荆苷对EC-109细胞凋亡的诱导作用|流式细胞术检测EC-109细胞中抑癌基因p53和促癌基因bcl-2蛋白的表达。结果牡荆苷对EC-109细胞生长、增殖具有明显的抑制作用,能够诱导EC-109细胞凋亡,且作用与牡荆苷浓度、作用时间呈正相关,可上调p53蛋白表达,下调bcl-2蛋白表达。结论牡荆苷可能通过上调p53蛋白和下调bcl-2蛋白的表达,促进EC-109细胞凋亡,抑制EC-109细胞生长。
[Key word]
[Abstract]
Objective To observe the effects of vitexin extracted from the flowers and flower buds of Trollius chinensis on the cell growth and apoptosis of human esophageal cancer EC-109 cells, and to investigate the effect of p53 and bcl-2 protein expression in antitumor mechanism. Methods EC-109 cells in logarithmic growth phase were treated by vitexin at different concentration. The inhibitory effects on the growth and proliferation of EC-109 cells in vitro were detected by CCK-8 method. The morphological changes of cell nucleus were assayed by the cell death detection kit Hoechst33258, DNA bands were detected by agarose gel electrophoresis, and the apoptosis rate of EC-109 cells induced by vitexin was examined using flow cytometry (FCM) with AnnexinV-FITC/PI double-labled technique. The protein expression of p53 and bcl-2 of EC-109 cells was detected by FCM. Results Vitexin had markedly inhibitory effect on the growth and proliferation of EC-109 cells, and the inhibitory rate increased with the concentration increasing and reaction time prolonging. Vitexin could induce the apoptosis of EC-109 cells and the protein expression of p53 was up-regulated while that of bcl-2 was down-regulated. Conclusion Vitexin could induce the apoptosis of EC-109 cells by up-regulating the expression of p53 and down-regulating that of bcl-2.
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[基金项目]
张家口市科技局资助项目(11110015D)