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

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Abstract

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.

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Acknowledgments

We acknowledge UGC-Major Research Project, New Delhi (F. No. 33-227/2007) and UGC-SAP for financial support. We would like to thank Dr. V. Jayanthi, DM. Former Head, Department of Medical Gastroenterology, Stanley Medical College and Hospital and Dr. Niranjali Devaraj Professor and Head, Department of Biochemistry, University of Madras, for their continuous support.

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Authors and Affiliations

  1. Unit of Biochemistry, Department of Zoology, School of Life Sciences, University of Madras, Guindy Campus, Chennai, 600 025, Tamil Nadu, India

    Natarajan Gopalakrishnan, Marimuthu Saravanakumar, Mani Thiyagu & Halagowder Devaraj

  2. Department of Biochemistry, University of Madras, Guindy Campus, Chennai, 600 025, Tamil Nadu, India

    Perumal Madankumar

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  1. Natarajan Gopalakrishnan
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Correspondence to Halagowder Devaraj.

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Gopalakrishnan, N., Saravanakumar, M., Madankumar, P. et al. Colocalization of β-catenin with Notch intracellular domain in colon cancer: a possible role of Notch1 signaling in activation of CyclinD1-mediated cell proliferation. Mol Cell Biochem 396, 281–293 (2014). https://doi.org/10.1007/s11010-014-2163-7

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