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07 September 2020 : Laboratory Research

Reduced Caudal Type Homeobox 2 (CDX2) Promoter Methylation Is Associated with Curcumin’s Suppressive Effects on Epithelial-Mesenchymal Transition in Colorectal Cancer Cells

Ting Chen1ABCDEF, Chun Yang2ABCDEFG*, Zhenli Xi3ABDEF, Fen Chen1ABCDE, Hailin Li2ABCDE

DOI: 10.12659/MSM.926443

Med Sci Monit 2020; 26:e926443

Abstract

BACKGROUND: Curcumin is a component of Curcuma longa with various biological activities. The present study aimed to investigate curcumin’s inhibitory effects on epithelial-mesenchymal transition (EMT) in colorectal cancer (CRC) cells and possible mechanisms of action underlying these effects.

MATERIAL AND METHODS: Human SW480 CRC cells were incubated with curcumin at 0.1, 0.2, 0.4, 0.8, or 1.6 μmol/L. The 3-(4,5-Dimethylthiazol-2-Yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay was utilized to evaluate cell viabilities. The DNA methylation levels of the cdx2 promoter were assessed by bisulfite sequencing polymerase chain reaction (BSP). Real-time quantitative PCR was used to measure the mRNA expression levels. Protein expression levels were evaluated with western blotting. Immunofluorescence staining was used to evaluate the nuclear translocation of β-catenin.

RESULTS: Curcumin concentrations of 0.1, 0.2, and 0.4 μmol/L showed no significant association with the viability of SW480 cells, which were chosen for subsequent experiments. Curcumin incubation significantly downregulated expression levels of DNA methyltransferase1 (DNMT1), DNMT3a, and the methylation levels of the cdx2 promoter in a concentration-dependent manner. The expression levels of N-cadherin, Vimentin, Wnt3a, Snail1, and Twist, as well as the nuclear translocation levels of β-catenin, were reduced in a curcumin concentration-dependent manner. The expression levels of E-cadherin were increased in a curcumin concentration-dependent manner.

CONCLUSIONS: Curcumin negatively regulated transcription factors promoting EMT in CRC cells by decreasing cdx2 promoter DNA methylation and consequently suppressing the CDX2/Wnt3a/β-catenin signaling pathway.

Keywords: Colorectal Neoplasms, Curcumin, Epithelial-mesenchymal transition, Wnt Signaling Pathway

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750