ObjectiveTo establish the malignant transformation cell model of colon cancer. MethodsImmortalized normal human colonic epithelial cells (NCM460 cells) were induced for multiple generations with N-methyl-n′-nitro-N-nitrosoguanidine (MNNG) as the initiator and phorbol 12-myristate 13-acetate (PMA) as the tumor promoter. The MTT assay, cell colony formation assay, and cell scratch assay were used to validate the malignant transformation tendency and malignant biological characteristics of NCM460 cells. The protein expressions of neural-cadherin (N-cadherin), transforming growth factor beta 1 (TGF-β1) and epidermal growth factor receptor (EGFR) were detected by the Western blot. ResultsThe morphology of normal NCM460 cells was spindle-shaped, arranged in order, and grew in a monolayer. After induction, the morphology of the 30th generation of NCM460 cells changed significantly, with different sizes, disordered arrangement, and clumped growth. The proliferation ability of normal NCM460 cells, the 17th generation of NCM460 cells and the 30th generation of NCM460 cells increased successively, the number of colony formation increased successively, and the migration distance and migration rate at 48 hours were increased or elevated successively (P＜0.05). The protein expressions of N-cadherin, EGFR and TGF-β1 in the 30th generation of NCM460 cells were higher than those in normal NCM460 cells (P＜0.05). ConclusionAfter continuous multiple generations of induction with MNNG and PMA, the biological functions (proliferation, colony formation, and migration) of NCM460 cells are significantly changed, and NCM460 cells have the characteristics of malignant cells. This model can be used to study the pathogenesis and therapeutic regimen of colon cancer.

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