高糖状态通过WIF1-Wnt/β-catenin通路调控结肠肿瘤细胞的增殖

High glucose regulates the proliferation of colon cancer cells through the WIF1-Wnt/β-catenin signaling pathway

目的分析高糖状态对结肠肿瘤细胞增殖的影响,并探讨Wnt抑制因子1(WIF1)通过Wnt/β-连环蛋白(β-catenin)通路影响高糖状态下结肠肿瘤细胞增殖的机制。方法用不同浓度的D-葡萄糖处理结肠肿瘤细胞株SW620细胞,通过实时荧光定量PCR(qPCR)和蛋白免疫印迹法测定增殖相关基因的表达,行细胞增殖活性检测、细胞计数实验,用细胞平板克隆形成实验检测细胞增殖情况,采用qPCR和蛋白免疫印迹法测定WIF1和β-catenin的表达。转染siRNA和过表达质粒调控WIF1的表达后,检测WIF1对SW620细胞增殖能力和β-catenin表达水平的影响。结果随着糖浓度的增加,SW620细胞的增殖能力增强(P均<0.05),WIF1的表达下降而β-catenin的表达增高(P均<0.05)。下调WIF1的表达,SW620细胞的增殖能力增强且β-catenin的表达增高(P均<0.05),而过表达WIF1后,SW620细胞的增殖能力受到抑制且β-catenin的表达下降(P均<0.05)。结论高糖状态下WIF1表达的下降,可能通过活化Wnt/β-catenin通路促进高糖状态下结肠肿瘤细胞的增殖。

Objective To evaluate the effect of high glucose on the proliferation of colon cancer cells,and investigate the mechanism of the effect of Wnt inhibitory factor 1(WIF1)on the proliferation of colon cancer cells under high glucose state through the Wnt/β-catenin signaling pathway.Methods Colon cancer cell line SW620 cells were treated with different doses of D-glucose.The expressions of proliferationassociated mRNA and protein were measured by real-time fluorescent quantitative PCR(qPCR)and Western blot.The proliferation ability of SW620 cells was measured by cell proliferation assay,cell counting assay and colony formation assay.The expression levels of WIF1 andβ-catenin were detected by qPCR and Western blot.After regulating the expression of WIF1 by the transfection with siRNA and over-expressed plasmid,the effect of WIF1 upon the proliferation ability and the expression ofβ-catenin in SW620 cells was evaluated.Results Along with the increase in glucose concentration,the proliferation ability of SW620 cells was significantly increased,the expression of WIF1 was remarkably down-regulated and the expression ofβ-catenin was significantly up-regulated(all P<0.05).The proliferation ability of SW620 cells was considerably increased and the expression ofβ-catenin was significantly up-regulated after down-regulating the WIF1 expression(all P<0.05).However,after the over-expression of WIF1,cell proliferation ability was significantly inhibited and the expression ofβ-catenin was remarkably down-regulated(all P<0.05).Conclusion Down-regulation of WIF1 expression under high glucose state may promote the proliferation of colon cancer cells under high glucose state via the activation of Wnt/β-catenin signaling pathway.