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Specific Smad2/3 Linker Phosphorylation Indicates Esophageal Non-neoplastic and Neoplastic Stem-Like Cells and Neoplastic Development

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Abstract

Background

There is little known about stem cells in human non-neoplastic and neoplastic esophageal epithelia. We have demonstrated expression of linker threonine-phosphorylated Smad2/3 (pSmad2/3L-Thr), suggesting presence of stem-like cells in mouse esophageal epithelium, and identified presence of pSmad2/3L-Thr-positive cells that might function as cancer stem cells in mouse model of colorectal carcinoma.

Aims

We explore whether pSmad2/3L-Thr can be used as a biomarker for stem cells of human esophageal epithelia and/or neoplasms.

Methods

We have used esophageal tissues from inpatients undergoing endoscopic submucosal dissection and performed double immunofluorescent staining of pSmad2/3L-Thr and Ki67, CDK4, p63, Sox2, CK14, p53, ALDH1, CD44 or D2-40 after which the sections were stained with hematoxylin and eosin.

Results

pSmad2/3L-Thr-positive cells showed immunohistochemical co-localization with CDK4, p63, CD44 and Sox2 in the basal and parabasal layers of non-neoplastic esophageal epithelia. In esophageal neoplasms, they showed immunohistochemical co-localization with p53, CDK4, ALDH1 and CD44. There was a significant increase in the percentage of pSmad2/3L-Thr-positive cells in the p53-positive neoplastic cell population with development of esophageal neoplasia. pSmad2/3L-Thr-positive cells localized to the lower section of low-grade intraepithelial neoplasia and were observed up to the upper section in carcinoma in situ. In invasive squamous cell carcinoma, they were scattered throughout the tumor with disappearance of polarity and were found in intraepithelial primary lesions and sites of submucosal and vessel invasion.

Conclusions

We determined significant expression of pSmad2/3L-Thr in human esophageal non-neoplastic and neoplastic epithelia, indicating that these are epithelial stem-like cells and cancer stem cells, respectively, that correlate with developing esophageal neoplasms.

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Acknowledgments

This study was supported by grant-in-aid for Research Grant (B) from Japanese Foundation for Research and Promotion of Endoscopy and grant-in-aid for Scientific Research (C) from Japan Society for the Promotion of Science (16K09330, 25460938).

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Authors and Affiliations

  1. Third Department of Internal Medicine, Division of Gastroenterology and Hepatology, Kansai Medical University, 2-5-1 Shinmachi, Hirakata, Osaka, 573-1010, Japan

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

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  1. Shunsuke Horitani
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  2. Toshiro Fukui
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Correspondence to Toshiro Fukui.

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All procedures performed in this study involving human participants were in accordance with the ethical standards of the institutional (number 2012101) and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.

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Horitani, S., Fukui, T., Tanimura, Y. et al. Specific Smad2/3 Linker Phosphorylation Indicates Esophageal Non-neoplastic and Neoplastic Stem-Like Cells and Neoplastic Development. Dig Dis Sci 66, 1862–1874 (2021). https://doi.org/10.1007/s10620-020-06489-8

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  • DOI: https://doi.org/10.1007/s10620-020-06489-8

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