‘MCC’ protein interacts with E-cadherin and β-catenin strengthening cell–cell adhesion of HCT116 colon cancer cells

Abstract

E-cadherin and β-catenin are key proteins that are essential in the formation of the epithelial cell layer in the colon but their regulatory pathways that are disrupted in cancer metastasis are not completely understood. Mutated in colorectal cancer (MCC) is a tumour suppressor gene that is silenced by promoter methylation in colorectal cancer and particularly in patients with increased lymph node metastasis. Here, we show that MCC methylation is found in 45% of colon and 24% of rectal cancers and is associated with proximal colon, poorly differentiated, circumferential and mucinous tumours as well as increasing T stage and larger tumour size. Knockdown of MCC in HCT116 colon cancer cells caused a reduction in E-cadherin protein level, which is a hallmark of epithelial-mesenchymal transition in cancer, and consequently diminished the E-cadherin/β-catenin complex. MCC knockdown disrupted cell-cell adhesive strength and integrity in the dispase and transepithelial electrical resistance assays, enhanced hepatocyte growth factor-induced cell scatter and increased tumour cell invasiveness in an organotypic assay. The Src/Abl inhibitor dasatinib, a candidate anti-invasive drug, abrogated the invasive properties induced by MCC deficiency. Mechanistically, we establish that MCC interacts with the E-cadherin/β-catenin complex. These data provide a significant advance in the current understanding of cell-cell adhesion in colon cancer cells.

Destruxin B Suppresses Drug-Resistant Colon Tumorigenesis and Stemness Is Associated with the Upregulation of miR-214 and Downregulation of mTOR/β-Catenin Pathway

“…In another experiment, we showed that a single DB pre-treatment (below IC50) could significantly reduce the tumor-initiating ability of IGF1-cultured DLD-1 colon CSC-like cells in vivo. Mechanistically, DB treatment was associated with the decreased expression of a key CSC marker, β-catenin, which has been shown to be aberrantly increased in malignant colon cancer cells and associated with the generation of drug-resistance [ 36 , 37 ]. DB-mediated suppression of β-catenin was also accompanied with an increased miR-214 level, which has a potential binding site at β-catenin’s 3′UTR.…”

Mutated in colorectal cancer (MCC) : a novel centrosomal protein involved in intestinal homeostasis and disease

“…Numerous published studies revealed the presence of MCC protein in assorted widely used cell lines by WB analysis. However, the precise subcellular localization of MCC was controversial with various conflicting reports (Arnaud et al, 2009;Benthani et al, 2018;Fukuyama et al, 2008;Matsumine et al, 1996;Pangon et al, 2010Pangon et al, , 2015Senda et al, 1997). This is due both to the dearth of published studies investigating (Fukuyama et al, 2008;Pangon et al, 2015).…”

“…Mcc is membrane associated and interacts with β-catenin and E-cadherin to strengthen cell adhesion in CRC cell lines (Benthani et al, 2018).…”

“…The MCC protein also contains several regions predicted to form coiled coil structures ( Figure 16B-C) (Bourne, 1991;Kinzler et al, 1991). In mouse and human, there are two isoforms of MCC (1 and 2) that are generated from alternative promoter usage and differ only at their N-termini ( Figure 10 (Arnaud et al, 2009;Benthani et al, 2018;Fukuyama et al, 2008;Li et al, 2013;Matsumine et al, 1996;Pangon et al, 2015).…”

“…We next aimed to reevaluate the subcellular localization of MCC in numerous cell lines including those known to contain endogenous MCC by Western blotting or in which MCC localization had been previously proposed (e.g., HEK293, SW480 and HCT116) (Benthani et al, 2018;Fukuyama et al, 2008). To this end, we surveyed numerous commercially available antibodies for MCC specificity and identified one that shows highly specific staining for endogenous MCC by IF and that works well in western blot analysis (Table 03).…”