Colocalization of β-catenin with Notch intracellular domain in colon cancer: a possible role of Notch1 signaling in activation of CyclinD1-mediated cell proliferation

• Published in: Molecular and cellular biochemistry 2014-11, Vol.396 (1-2), p.281-293
• Main Authors: Gopalakrishnan, Natarajan, Saravanakumar, Marimuthu, Madankumar, Perumal, Thiyagu, Mani, Devaraj, Halagowder
• Format: Article
• Language: English
• Subjects: Adenocarcinoma - metabolism; Adenocarcinoma - pathology; Adenoma - metabolism; Adenoma - pathology; Basic Helix-Loop-Helix Transcription Factors - genetics; beta Catenin - metabolism; Biochemistry; Biomedical and Life Sciences; Cardiology; Cell Proliferation; Colonic Neoplasms - genetics; Colonic Neoplasms - metabolism; Colonic Neoplasms - pathology; Colorectal Neoplasms - genetics; Colorectal Neoplasms - metabolism; Colorectal Neoplasms - pathology; Cyclin D1 - genetics; Cyclin D1 - metabolism; Dipeptides - pharmacology; Dose-Response Relationship, Drug; Gene Expression Regulation, Neoplastic; Homeodomain Proteins - genetics; HT29 Cells - drug effects; HT29 Cells - metabolism; Humans; Life Sciences; Medical Biochemistry; Mucin-2 - genetics; Mucin-2 - metabolism; Oncology; Protein Structure, Tertiary; Receptor, Notch1 - metabolism; Reference Values; Signal Transduction; Transcription Factor HES-1
• ISSN: 0300-8177; 1573-4919
• DOI: 10.1007/s11010-014-2163-7

The Wnt and Notch1 signaling pathways play major roles in intestinal development and tumorigenesis. Sub-cellular localization of β-catenin has been implicated in colorectal carcinogenesis. However, the β-catenin and Notch intracellular domain (NICD) interaction has to be addressed. Immunohistochemistries of β-catenin, NICD, and dual immunofluorescence of β-catenin and NICD were analyzed in colorectal tissues and HT29 cell line. Moreover, real-time PCR analysis of CyclinD1, Hes1 and MUC2 was done in HT29 cells upon N -[ N -(3, 5-difluorophenacetyl)- l -alanyl]- S -phenylglycine t -butyl ester (DAPT) treatment. Dual staining emphasized the strong interaction of β-catenin and NICD in adenoma and adenocarcinoma than in normal tissues. Hes1 transcript levels were decreased 1.5- and 7.1-fold in 12.5 and 25 µM DAPT-treated HT29 cells. CyclinD1 transcript levels decreased 1.2- and 1.6-fold, and MUC2 transcript level increased 4.3- and 7.5-fold in 12.5 and 25 µM DAPT-treated HT29 cells. The results of this study showed that the sub-cellular localization of β-catenin converges with NICD inducing proliferation through the activation of CyclinD1 and Hes1. Moreover, the inhibition of Notch1 signaling by DAPT leads to the arrest of cell proliferation and induces apoptosis leading to the upregulation of MUC2, a secretory cell lineage marker.