Gasdermin C Is Upregulated by Inactivation of Transforming Growth Factor β Receptor Type II in the Presence of Mutated Apc, Promoting Colorectal Cancer Proliferation

Masashi Miguchi; Takao Hinoi; Manabu Shimomura; Tomohiro Adachi; Yasufumi Saito; Hiroaki Niitsu; Masatoshi Kochi; Haruki Sada; Yusuke Sotomaru; Tsuneo Ikenoue; Kunitoshi Shigeyasu; Kohji Tanakaya; Yasuhiko Kitadai; Kazuhiro Sentani; Naohide Oue; Wataru Yasui; Hideki Ohdan

doi:10.1371/journal.pone.0166422

Gasdermin C Is Upregulated by Inactivation of Transforming Growth Factor β Receptor Type II in the Presence of Mutated Apc, Promoting Colorectal Cancer Proliferation

PLoS One. 2016 Nov 11;11(11):e0166422. doi: 10.1371/journal.pone.0166422. eCollection 2016.

Authors

Masashi Miguchi¹, Takao Hinoi^{1

2}, Manabu Shimomura¹, Tomohiro Adachi¹, Yasufumi Saito¹, Hiroaki Niitsu¹, Masatoshi Kochi¹, Haruki Sada¹, Yusuke Sotomaru³, Tsuneo Ikenoue⁴, Kunitoshi Shigeyasu⁵, Kohji Tanakaya⁵, Yasuhiko Kitadai^{6

7}, Kazuhiro Sentani⁸, Naohide Oue⁸, Wataru Yasui⁸, Hideki Ohdan¹

Affiliations

¹ Department of Gastroenterological and Transplant Surgery, Applied Life Sciences, Institute of Biomedical & Health Sciences, Hiroshima University, Hiroshima, Japan.
² Department of Surgery, Institute for Clinical Research, National Hospital Organization Kure Medical Center and Chugoku Cancer Center, Hiroshima, Japan.
³ Natural Science Center for Basic Research and Development, Hiroshima University, Hiroshima, Japan.
⁴ Division of Clinical Genome Research, the Institute of Medical Science, University of Tokyo, Tokyo, Japan.
⁵ Department of Surgery, National Hospital Organization Iwakuni Clinical Center, Yamaguchi, Japan.
⁶ Departments of Gastroenterology and Metabolism, Applied Life Sciences, Institute of Biomedical & Health Sciences, Hiroshima University, Hiroshima, Japan.
⁷ Department of Health Science, Program in Human Culture and Science, Prefectural University of Hiroshima, Hiroshima, Japan.
⁸ Department of Molecular Pathology, Applied Life Sciences, Institute of Biomedical & Health Sciences, Hiroshima University, Hiroshima, Japan.

Abstract

Mutations in TGFBR2, a component of the transforming growth factor (TGF)-β signaling pathway, occur in high-frequency microsatellite instability (MSI-H) colorectal cancer (CRC). In mouse models, Tgfbr2 inactivation in the intestinal epithelium accelerates the development of malignant intestinal tumors in combination with disruption of the Wnt-β-catenin pathway. However, no studies have further identified the genes influenced by TGFBR2 inactivation following disruption of the Wnt-β-catenin pathway. We previously described CDX2P-G19Cre;Apcflox/flox mice, which is stochastically null for Apc in the colon epithelium. In this study, we generated CDX2P-G19Cre;Apcflox/flox;Tgfbr2flox/flox mice, with simultaneous loss of Apc and Tgfbr2. These mice developed tumors, including adenocarcinoma in the proximal colon. We compared gene expression profiles between tumors of the two types of mice using microarray analysis. Our results showed that the expression of the murine homolog of GSDMC was significantly upregulated by 9.25-fold in tumors of CDX2P-G19Cre;Apcflox/flox;Tgfbr2flox/flox mice compared with those of CDX2P-G19Cre;Apcflox/flox mice. We then investigated the role of GSDMC in regulating CRC tumorigenesis. The silencing of GSDMC led to a significant reduction in the proliferation and tumorigenesis of CRC cell lines, whereas the overexpression of GSDMC enhanced cell proliferation. These results suggested that GSDMC functioned as an oncogene, promoting cell proliferation in colorectal carcinogenesis. In conclusion, combined inactivation of both Apc and Tgfbr2 in the colon epithelium of a CRC mouse model promoted development of adenocarcinoma in the proximal colon. Moreover, GSDMC was upregulated by TGFBR2 mutation in CRC and promoted tumor cell proliferation in CRC carcinogenesis, suggesting that GSDMC may be a promising therapeutic target.

MeSH terms

Adenocarcinoma / genetics*
Adenocarcinoma / metabolism
Adenocarcinoma / pathology
Adenomatous Polyposis Coli Protein / antagonists & inhibitors
Adenomatous Polyposis Coli Protein / genetics*
Adenomatous Polyposis Coli Protein / metabolism
Animals
Biomarkers, Tumor / antagonists & inhibitors
Biomarkers, Tumor / genetics*
Biomarkers, Tumor / metabolism
CDX2 Transcription Factor / genetics
CDX2 Transcription Factor / metabolism
Cell Line, Tumor
Cell Proliferation
Colon / metabolism
Colon / pathology
Colorectal Neoplasms / genetics*
Colorectal Neoplasms / metabolism
Colorectal Neoplasms / pathology
DNA-Binding Proteins / antagonists & inhibitors
DNA-Binding Proteins / genetics*
DNA-Binding Proteins / metabolism
Female
Gene Expression Regulation, Neoplastic*
Humans
Male
Mice
Mice, Transgenic
Microarray Analysis
Mutation
Protein Serine-Threonine Kinases / antagonists & inhibitors
Protein Serine-Threonine Kinases / deficiency
Protein Serine-Threonine Kinases / genetics*
RNA, Small Interfering / genetics
RNA, Small Interfering / metabolism
Receptor, Transforming Growth Factor-beta Type II
Receptors, Transforming Growth Factor beta / antagonists & inhibitors
Receptors, Transforming Growth Factor beta / deficiency
Receptors, Transforming Growth Factor beta / genetics*
Signal Transduction
Transforming Growth Factor beta / genetics
Transforming Growth Factor beta / metabolism

Substances

Adenomatous Polyposis Coli Protein
Biomarkers, Tumor
CDX2 Transcription Factor
DNA-Binding Proteins
GSDMC protein, human
RNA, Small Interfering
Receptors, Transforming Growth Factor beta
Transforming Growth Factor beta
adenomatous polyposis coli protein, mouse
Protein Serine-Threonine Kinases
Receptor, Transforming Growth Factor-beta Type II

Grants and funding

This work was supported by the Japan Society for the Promotion of Science (JSPS) KAKENHI Grant-in-Aid for Scientific Research (http://www.jsps.go.jp/j-grantsinaid/) (Grant Numbers 22390257 and 25293284) to TH, the Japan Society for the Promotion of Science (JSPS) KAKENHI Grant-in-Aid for young Scientists Research (http://www.jsps.go.jp/j-grantsinaid/) (Grant Numbers 25861190 and 15K19892) to MS, the Japanese Society of Gastroenterology Grant-in-Aid 2010, and the Nakayama Cancer Research Institute Grant-in-Aid 2009 for Gastrointestinal Disease. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.