姜黄素对人肺鳞癌细胞NCI-H292黏蛋白5AC表达的影响及机制

Effect of curcumin on expression of mucin 5AC in human lung squamous cell carcinoma cell line NCI-H292 and its mechanism

目的观察姜黄素对人肺鳞癌细胞NCI-H292黏蛋白5AC(MUC5AC)表达的影响,并探讨其机制。方法将细胞分为姜黄素和对照组,分别加入姜黄素、等量DMSO干预,Western blotting法检测MUC5AC、叉头框蛋白A2(FOXA2)和磷酸化信号转导及转录激活蛋白6(p-STAT6);将细胞分为FOXA2 siRNA组和NC-siRNA组,分别给予FOXA2 siRNA、negative-siRNA转染24 h,采用Western blotting法检测MUC5AC、FOXA2;将转染后的两组细胞分别予以姜黄素、等量DMSO孵育48 h,采用Western blotting法检测MUC5AC。将细胞分为对照组、抑制剂组、姜黄素组,分别予以DMSO、STAT6抑制剂Kaempferol及姜黄素处理48 h,采用RT-PCR法检测FOXA2 mRNA。结果与对照组比较,姜黄素组MUC5AC、p-STAT6表达降低而FOXA2表达增高(P均<0. 01)。与NC-siRNA组比较,FOXA2 siRNA组FOXA2表达水平降低而MUC5AC表达增高(P均<0. 01);与同组DMSO作用的细胞比较,NCsiRNA组、FOXA2 siRNA组加入姜黄素后MUC5AC表达降低(P均<0. 01)。FOXA2 mRNA相对表达量对照组<抑制剂组<姜黄素组(P均<0. 01)。结论姜黄素通过抑制STAT6磷酸化入核上调FOXA2表达,从而抑制人肺鳞癌NCI-H292细胞分泌MUC5AC。

Objective To investigate the effect of curcumin on the expression of mucin 5AC(MUC5AC)in human lung squamous cell carcinoma cell line NCI-H292 and its mechanism.Methods The cells were divided into two groups:the curcumin group and control group,and curcumin and DMSO were added,respectively.The expression levels of MUC5AC,forkhead box protein A2(FOXA2)and phosphorylation signal transduction and activator protein 6(p-STAT6)were detected by Western blotting;the cells were divided into two groups:FOXA2 siRNA group and NC-siRNA group,which were transfected with FOXA2 siRNA and negative-siRNA for 24 h,respectively.The expression levels of MUC5AC and FOXA2 were detected by Western blotting.The transfected two groups were treated with curcumin and the same amount of DMSO for 48 h,respectively,and we detected MUC5AC by Western blotting.The cells were divided into the control group,inhibitor group and curcumin group,which were treated with DMSO,STAT6 inhibitor Kaempferol,and curcumin for 48 h,respectively.The FOXA2 mRNA was detected by RT-PCR.Results Compared with the control group,the expression of MUC5AC and p-STAT6 decreased and the expression of FOXA2 increased in the curcumin group(all P<0.01).Compared with the NC-siRNA group,FOXA2 expression level decreased and MUC5AC protein expression increased in the FOXA2 siRNA group(both P<0.01).Compared with the cells treated with DMSO,the expression of MUC5AC decreased after adding curcumin in the NC-siRNA group and FOXA2 siRNA group(all P<0.01).The relative expression of FOXA2 mRNA was as follows:control group<inhibitor group<curcumin group(P<0.01).Conclusion Curcumin up-regulates the FOXA2 protein expression by inhibiting STAT6 phosphorylation into the nucleus,thereby inhibiting the secretion of MUC5AC by NCI-H292 cells.