姜黄素对人口腔黏膜上皮细胞系增殖及Wnt/β-catenin信号通路的影响

Effects of curcumin on proliferation and Wnt/β-catenin signaling pathway of human oral mucosal epithelial cell lines

目的:探究姜黄素对人口腔黏膜上皮细胞系增殖及Wnt/β-catenin信号通路的影响。方法:取生长至对数期的人口腔上皮癌Ca9-22细胞,分为对照组及5μmol/L姜黄素处理组、10μmol/L姜黄素处理组、20μmol/L姜黄素处理组、40μmol/L姜黄素处理组,分别于处理24、48、72 h后采用MTT法检测细胞存活率,采用流式细胞仪检测不同浓度姜黄素对Ca9-22细胞凋亡的影响,Caspase-3、Caspase-9活性检测试剂盒检测Caspase-3、Caspase-9活性,免疫印迹法检测β-catenin、C-myc、Wnt1、Bcl-2及Bax表达。结果:与对照组比较,不同浓度姜黄素均对人口腔上皮癌Ca9-22细胞的增殖有一定抑制作用,且与浓度、时间有关。5μmol/L姜黄素处理组、10μmol/L姜黄素处理组、20μmol/L姜黄素处理组、40μmol/L姜黄素组处理48 h后,细胞凋亡率、Caspase-3、Caspase-9活性及Bax mRNA表达均高于对照组,Bcl-2 mRNA及β-catenin、C-myc、Wnt1表达低于对照组,差异有统计学意义(均P<0.05)。结论:姜黄素可抑制人口腔上皮癌Ca9-22细胞增殖并诱导Ca9-22细胞凋亡,其机制与抑制Wnt/β-catenin信号通路的表达有关。

Objective:To investigate effect of curcumin on the proliferation and Wnt/β-catenin signaling pathway of human oral mucosa epithelial cell lines.Methods:Human oral epithelial carcinoma Ca9-22 cells grown to logarithmic stage were collected and divided into control group and 5μmol/L curcumin group,10μmol/L curcumin group,20μmol/L curcumin group,40μmol/L curcumin group.After 24,48,72 h of treatment,the cell viability was detected by MTT method,the effect of different concentrations of curcumin on the apoptosis of Ca9-22 cells was detected by flow cytometry,and the Caspase-3 and Caspase-9 activity detection kits were used to detect Caspase-3 Caspase-9 activity,the expression ofβ-catenin,C-myc,Wnt1,Bcl-2 and Bax were detected by Western blot.Results:Compared with the control group,different concentrations of curcumin had certain inhibitory effect on the proliferation of human oral epithelial cancer Ca9-22 cells,and the inhibitory effect was related to the concentration and time.The apoptosis rate and Caspase-3,Caspase-9 activity,Bax mRNA expression in 5μmol/L curcumin group,10μmol/L curcumin group,20μmol/L curcumin group,40μmol/L curcumin group were higher than those in the control group after 48 hours of treatment.The expressions of Bcl-2 mRNA,β-catenin,C-myc and Wnt1 were significantly lower than those in the control group,difference statistically significant(all P<0.05).Conclusion:Curcumin can inhibit the proliferation of human oral epithelial cancer Ca9-22 cells,induce apoptosis of Ca9-22 cells.The mechanism is related to the inhibition of the activation of Wnt/β-catenin signaling pathway.