摘要
目的:探讨黄腐酚通过Notch信号通路对甲状腺癌B-CPAP细胞增殖及凋亡影响.方法:体外培养甲状腺癌B-CPAP细胞,以不同浓度黄腐酚干预培养.根据黄腐酚浓度不同分为对照组(0 μ mol/L)、低剂量组(10 μmol/L)、中剂量组(20 μmol/L)、高剂量组(40 μ mol/L).采用MTT法检测黄腐酚对B-CPAP细胞的增殖抑制率.采用Hoechst 33258染色方法,应用荧光显微镜观察不同组别B-CPAP细胞形态的变化.采用Annexin V-FITC/PI双染法流式细胞术检测B-CPAP细胞凋亡率.采用West-ern-blot 法测定Notch信号通路相关蛋白变化.结果:MTT检测结果表明,低剂量组、中剂量组及高剂量组分别处理24、48、72 h后,细胞增殖抑制率差异均有统计学意义(P＜0.05);同剂量组处理24、48、72 h后,细胞增殖抑制率逐渐增高,差异有统计学意义(F=204.51、1 69.64、183.15,P＜0.05).与对照组比较,高剂量组荧光显微镜下显示出典型细胞凋亡的形态学改变.流式细胞术检测B-CPAP细胞经不同剂量黄腐酚处理72 h后,凋亡率比较差异有统计学意义(F=1235.54,P＜0.05),高剂量组凋亡率更高.Western-blot法结果显示,处理72 h,与对照组比较,低剂量组、中剂量组、高剂量组Notch1、Hes1、Bcl-2蛋白表达明显降低,而Cleaved caspase-3表达明显增高,差异均有统计学意义(P＜0.05).结论:黄腐酚可能通过Notch信号通路抑制B-CPAP细胞增殖及诱导凋亡.
Abstract
Objective:Xanthohumol is a kind of isoamyl olefinic flavonoid natural compounds,which have antitumor activity and impact on a variety of cell signaling pathways,The objective of this study was to explore the effects of xanthohumol on proliferation and apoptosis of thyroid cancer cells B-CPAP through the Notch signaling pathway.Methods:B-CPAP cells were cultivated in vitro,Xanthohumol was divided into control group(0 μ mol/L),low dose group(10 μ mol/L),middle dose group(20 μ mol/L),high dose group(40 μ mol/L)according to the different concentrations,The logarithmic growth cells were cultivated with different concentrations of xanthohumol intervention,application of MTT colorimetry in the detection of proliferation inhibition rates of B-CPAP cells.B-CPAP cells morphological changes were observed by using fluorescence microscope after appli-cation of Hoechst 33258 dyeing.B-CPAP cells apoptosis were detected by Annexin V-FITC/PI flow cytometry.Notch signaling pathway related proteins were determined?by?Western blotting.Re-sults:MTT showed that low dose group,middle dose group and high dose group,respectively pro-cessing after 24h,48h,72h,proliferation inhibition rates of the three groups were statistical?signifi-cance(F=189.34,131.73,124.51,P＜0.05);Respectively treated after 24h,48h,72h,proliferation in-hibition rates of xanthohumol increased over time in the same group,The differences were statisti-cally significant(F=204.51,169.64,183.15,P＜0.05).B-CPAP cells of high dose group appeared ob-viously apoptosis morphological changes compared with the control group through Hoechst33258 dying.Flow cytometry showed apoptosis rates of concrol group,low dose group and high dose group compared were statistical?significance(F=1235.54,P＜0.05).Apoptosis rate was higher in the high-dose group.Western blotting showed that Notch1,Treatment was performed for 72h,Hes1,Bcl-2 expression were significantly decreased in low dose group,middle dose group and high dose group compared with the control group(F=203.22,161.52,224.78),while cleaved caspase-3 ex-pression significantly increased(F=463.27),the differences were statistically significant(P＜0.05).Conclusions:Xanthohumol inhibits B-CPAP cells proliferation and induces cells apoptosis maybe through the Notch signaling pathway.
基金项目
济南市卫生健康委科技项目(第二批)(2020)(2020-3-54)
维普
万方数据