黄腐酚通过Notch信号通路对甲状腺癌细胞B-CPAP增殖及凋亡的影响

Effects of xanthohumol on proliferation and apoptosis of thyroid cancer cells B-CPAP through the Notch signaling pathway

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(0μmol/L)、低剂量组(10μmol/L)、中剂量组(20μmol/L)、高剂量组(40μmol/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Western-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、169.64、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。

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(0μmol/L),low dose group(10μmol/L),middle dose group(20μmol/L),high dose group(40μmol/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 application 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.Results:MTT showed that low dose group,middle dose group and high dose group,respectively processing after 24h,48h,72h,proliferation inhibition rates of the three groups were statistical?significance(F=189.34,131.73,124.51,P<0.05);Respectively treated after 24h,48h,72h,proliferation inhibition rates of xanthohumol increased over time in the same group,The differences were statistically significant(F=204.51,169.64,183.15,P<0.05).B-CPAP cells of high dose group appeared obviously 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 expression 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.