Myosin light chain kinase expression induced via tumor necrosis factor receptor 2 signaling in the epithelial cells regulates the development of colitis-associated carcinogenesis

Masahiro Suzuki; Takashi Nagaishi; Motomi Yamazaki; Michio Onizawa; Taro Watabe; Yuriko Sakamaki; Shizuko Ichinose; Mamoru Totsuka; Shigeru Oshima; Ryuichi Okamoto; Motoyuki Shimonaka; Hideo Yagita; Tetsuya Nakamura; Mamoru Watanabe

doi:10.1371/journal.pone.0088369

Myosin light chain kinase expression induced via tumor necrosis factor receptor 2 signaling in the epithelial cells regulates the development of colitis-associated carcinogenesis

PLoS One. 2014 Feb 10;9(2):e88369. doi: 10.1371/journal.pone.0088369. eCollection 2014.

Affiliations

¹ Department of Gastroenterology, Graduate School of Medical Science, Tokyo Medical and Dental University, Tokyo, Japan.
² Department of Obstetrics and Gynecology, Nihon University School of Medicine, Tokyo, Japan.
³ Research Center for Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
⁴ Department of Applied Biological Chemistry, The University of Tokyo, Tokyo, Japan.
⁵ Department of Chemistry, Tokyo University of Science, Tokyo, Japan.
⁶ Department of Immunology, Juntendo University School of Medicine, Tokyo, Japan.

Abstract

It has been suggested that prolonged inflammatory bowel diseases (IBD) may lead to colitis-associated carcinogenesis (CAC). We previously observed that the NF-κB activation in colonic epithelial cells is associated with increased tumor necrosis factor receptor 2 (TNFR2) expression in CAC development. However, the mechanism by which epithelial NF-κB activation leading to CAC is still unclear. Myosin light chain kinase (MLCK) has been reported to be responsible for the epithelial permeability associated with TNF signaling. Therefore we focused on the role of MLCK expression via TNFR2 signaling on CAC development. Pro-tumorigenic cytokines such as IL-1β, IL-6 and MIP-2 production as well as INF-γ and TNF production at the lamina propria were increased in the setting of colitis, and further in tumor tissues in associations with up-regulated TNFR2 and MLCK expressions in the epithelial cells of a CAC model. The up-regulated MLCK expression was observed in TNF-stimulated colonic epithelial cells in a dose-dependent fashion in association with up-regulation of TNFR2. Silencing TNFR2, but not TNFR1, resulted in restoration of epithelial tight junction (TJ) associated with decreased MLCK expression. Antibody-mediated blockade of TNF signaling also resulted in restoration of TJ in association with suppressed MLCK expression, and interestingly, similar results were observed with suppressing TNFR2 and MLCK expressions by inhibiting MLCK in the epithelial cells. Silencing of MLCK also resulted in suppressed TNFR2, but not TNFR1, expression, suggesting that the restored TJ leads to reduced TNFR2 signaling. Such suppression of MLCK as well as blockade of TNFR2 signaling resulted in restored TJ, decreased pro-tumorigenic cytokines and reduced CAC development. These results suggest that MLCK may be a potential target for the prevention of IBD-associated tumor development.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Carcinogenesis / drug effects
Carcinogenesis / metabolism
Carcinogenesis / pathology*
Cell Line
Cell Proliferation / drug effects
Colitis / metabolism
Colitis / pathology*
Colon / drug effects
Colon / pathology
Colon / ultrastructure
Cytokines / metabolism
Disease Models, Animal
Epithelial Cells / enzymology*
Epithelial Cells / metabolism
Epithelial Cells / pathology
Epithelial Cells / ultrastructure
Female
Inflammation / pathology
Inflammation Mediators / metabolism
Interferon-gamma / pharmacology
Intestinal Mucosa / drug effects
Intestinal Mucosa / pathology
Mice
Mice, Inbred BALB C
Mice, Inbred C57BL
Myosin-Light-Chain Kinase / metabolism*
Receptors, Tumor Necrosis Factor, Type II / metabolism*
Signal Transduction* / drug effects
Tight Junctions / drug effects
Tight Junctions / metabolism
Tumor Necrosis Factor-alpha / pharmacology
Up-Regulation / drug effects

Substances

Cytokines
Inflammation Mediators
Receptors, Tumor Necrosis Factor, Type II
Tumor Necrosis Factor-alpha
Interferon-gamma
Myosin-Light-Chain Kinase

Grants and funding

This study was supported in part by Grants-in-Aid for Scientific Research (T. Nagaishi, T. Nakamura and MW), Scientific Research on Priority Areas (MW), Exploratory Research and Creative Scientific Research (MW) from the Japanese Ministry of Education, Culture, Sports, Science and Technology; the Japanese Ministry of Health, Labor and Welfare (MW); the Japan Medical Association (MW); Japan Foundation for Applied Enzymology (MO); Japan Health Sciences Foundation (MO); Memorial Fund of Nihon Univ. Medical Alumni Association (T. Nagaishi); Abbott Japan Allergy Research Award (T. Nagaishi); Foundation for Advancement of International Science (T. Nagaishi); Takeda Science Foundation (T. Nagaishi). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.