Over-expression of SCUBE2 suppresses epithelial-mesenchymal transition through Wnt/β-catenin signaling pathway in colorectal cancer cells / 中国病理生理杂志

ABSTRACT

AIM: To study the effect of SCUBE2 on epithelial-mesenchymal transition (EMT) in colorectal cancer cells and its mechanism .METHODS:The expression of SCUBE2 in human colorectal cancer cell line HCT 116 and normal colonic cell line FHC was detected by real-time PCR and Western blot .HCT116 cells were transfected with GV144-SCUBE2 to over-express SCUBE2, and then the cell viability, migration, and apoptosis were determined by MTT assay , Transwell assay and flow cytometry, respectively.The expression of EMT markers (E-cadherin, vimentin, and Snail),β-catenin, c-Myc and cyclin D1 in the HCT116 cells was analyzed by real-time PCR or Western blot after transfection with GV144-SCUBE2 for 6 h, followed by the stimulation of 10 μg/L recombinant TGF-β1 protein for 48 h.Additionally, the EMT process of HCT116 cells, which were stimulated by TGF-β1, over-expressed SCUBE2, and treated with Wnt/β-cate-nin pathway activator lithium chloride ( LiCl) or inhibitor XAV93920, was analyzed by Western blot .RESULTS: Com-pared with FHC cells , the expression of SCUBE 2 in the HCT116 cells was significantly decreased .The viability and migra-tion ability of the HCT116 cells were suppressed by SCUBE2 over-expression, but the apoptosis was not markedly changed . Elevated expression of SCUBE2 increased E-cadherin expression, and decreased the expression of vimentin, Snail,β-cate-nin, c-Myc and cyclin D1 induced by TGF-β1.Treatment with LiCl blocked but treatment with XAV 93920 enhanced the effects of SCUBE2 on EMT.CONCLUSION:Over-expression of SCUBE2 may inhibit the cell growth and migration , and suppress EMT through Wnt/β-catenin signaling pathway .