Long‑term model of colitis‑associated colorectal cancer suggests tumor spread mechanism and nature of cancer stem cells

Tanimura,Yuji; Fukui,Toshiro; Horitani,Shunsuke; Matsumoto,Yasushi; Miyamoto,Sachi; Suzuki,Ryo; Tanaka,Toshihiro; Tomiyama,Takashi; Ikeura,Tsukasa; Ando,Yugo; Nishio,Akiyoshi; Okazaki,Kazuichi

doi:10.3892/ol.2020.12268

Long‑term model of colitis‑associated colorectal cancer suggests tumor spread mechanism and nature of cancer stem cells

Authors:
- Yuji Tanimura
- Toshiro Fukui
- Shunsuke Horitani
- Yasushi Matsumoto
- Sachi Miyamoto
- Ryo Suzuki
- Toshihiro Tanaka
- Takashi Tomiyama
- Tsukasa Ikeura
- Yugo Ando
- Akiyoshi Nishio
- Kazuichi Okazaki
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Affiliations: Third Department of Internal Medicine, Division of Gastroenterology and Hepatology, Kansai Medical University, Hirakata, Osaka 573‑1010, Japan
Published online on: November 3, 2020 https://doi.org/10.3892/ol.2020.12268
Article Number: 7

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Abstract

Although chemical‑induced animal models of colorectal cancer (CRC) suggest a lot about the disease, more efforts are required to establish metastasis models. Azoxymethane (AOM) and dextran sodium sulfate (DSS)‑treated (AOM/DSS) Crl:CD‑1 mice were sacrificed after 10 or 20 weeks in our previous study, and most colon tumors exhibited intramucosal adenocarcinomas. Our observations were extended until 30 weeks to study a colitis‑associated advanced CRC mouse model, and explore whether linker threonine‑phosphorylated Smad2/3 (pSmad2/3L‑Thr) immunostaining‑positive cells were involved in the progressive course of colitis‑associated CRC as cancer stem cells. AOM/DSS mice were sacrificed at 10, 20 and 30 weeks after AOM administration. Following the histopathological analysis, immunohistochemical staining was performed for the following markers: CD34, podoplanin, β‑catenin, E‑cadherin, Ki67, Bmi1 and pSmad2/3L‑Thr. Compared with AOM/DSS mice at 10 and 20 weeks, submucosal tumor infiltration and tumor invasion into vessels were markedly increased at 30 weeks. In the parts of colon tumors from AOM/DSS mice, particularly in mice at 30 weeks, the positive signal of E‑cadherin was clearly reduced in the cell membranes. The percentage of Ki67‑positive tumor cells in mucosal areas of AOM/DSS mice was higher than that in the sites of submucosal infiltration. In mucosal areas of colon tumors, pSmad2/3L‑Thr‑positive cells were scattered among tumor cells. At sites of submucosal infiltration and vessel invasion of these tumors, pSmad2/3L‑Thr‑positive cells were also observed among tumor cells. In colon tumors from AOM/DSS mice at 30 weeks, the percentage of pSmad2/3L‑Thr‑positive cells among the nuclear β‑catenin‑positive tumor cells was higher than that among the cytoplasmic β‑catenin‑positive tumor cells. For both non‑neoplastic and neoplastic epithelial cells, pSmad2/3L‑Thr‑positive cells exhibited immunohistochemical co‑localization with Bmi1. The present study developed an advanced CRC mouse model that exhibited tumor infiltration into the submucosa and invasion into vessels. The present study re‑confirmed the theory that pSmad2/3L‑Thr‑positive cells may be cancer stem cells.

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