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Identification of marker genes and pathways specific to precancerous duodenal adenomas and early stage adenocarcinomas

  • Original Article—Alimentary Tract
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Abstract

Background

The mechanism behind the pathogenesis and carcinogenesis of these neoplasms is not fully understood. The objective of this study was to identify genetic markers and pathways specific to precancerous duodenal adenomas and early stage adenocarcinomas through gene expression analysis.

Methods

Gene expression profiling was performed in 4 pairs of duodenal adenoma/adenocarcinomas and corresponding matched normal tissue. Genes with consistent expression differences were identified and confirmed in 7 independent pairs. Gene set enrichment analysis (GSEA) was performed to characterize gene expression profiles of duodenal adenoma/adenocarcinomas, together with immunohistochemical staining of candidate oncogenic genes.

Results

626 probes consistently demonstrated over a twofold expression difference between tumor–normal pairs. Reverse transcriptase polymerase chain reaction of genes with the most prominent difference in expression between tumors and normal mucosa (KLK7, KLK6, CEMIP, MMP7, KRT17, LGR5, G6PC, S100G, APOA1) validated the results of gene expression analysis. GSEA demonstrated a strong association between duodenal adenoma/adenocarcinomas with colorectal adenomas (p < 10−5) and gene expression patterns seen after APC gene knockout (p < 10−5), suggesting that the Wnt/β-catenin pathway plays a crucial role in the carcinogenesis of these neoplasms. Immunohistochemical staining of an independent group of duodenal adenomas confirmed over-accumulation of β-catenin in 80.0% (16/20).

Conclusions

Precancerous duodenal adenomas and early stage adenocarcinomas demonstrate gene expression characteristics with a strong resemblance to colorectal adenomas. The results of this study strongly suggest that upregulation of the Wnt/β-catenin pathway is the major factor involved in the initial stages of the carcinogenesis of duodenal adenocarcinomas.

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Acknowledgements

The authors thank Shinya Kodashima, Satoshi Ono, Yosuke Tsuji, Keiko Niimi for their assistance as participating investigators in acquisition of samples, and Hironori Waki, Yuta Hiraike for their assistance in RNA extraction.

Funding

This work was supported in part by a research grant from the Japanese Foundation for Research and Promotion of Endoscopy, in part by Grant-in-Aid for Young Scientists (B) from the Japan Society for the Promotion of Science, and in part by Grant-in-Aid for Scientific Research (C) from the Japan Society for the Promotion of Science.

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Authors and Affiliations

  1. Department of Gastroenterology, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8655, Japan

    Yoshiki Sakaguchi, Nobutake Yamamichi, Chihiro Takeuchi, Natsuko Kageyama-Yahara, Yu Takahashi, Mitsuhiro Fujishiro & Kazuhiko Koike

  2. Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, 2-5-1 Shikata-cho, Okayama, Okayama, Japan

    Shuta Tomida

  3. Department of Pathology, Fujita Health University School of Medicine, 1-98 Dengakugakubo, Kutsukake-cho, Toyoake, Aichi, Japan

    Kazuya Shiogama & Ken-ichi Inada

  4. Second Department of Internal Medicine, Wakayama Medical University, 811-1 Kimiidera, Wakayama, Japan

    Masao Ichinose

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  1. Yoshiki Sakaguchi
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  2. Nobutake Yamamichi
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Contributions

Conception and design: YS, NY, ST. Development of methodology: YS, NY, ST, CT, NKY, YT, KS, KI. Acquisition of data: YS, MI, MF. Analysis and interpretation of data: YS, NY, ST, CT, YT, KS, KI. Writing, review, and/or revision of the manuscript: YS, NY, ST, CT, NKY, YT, KK.

Corresponding author

Correspondence to Nobutake Yamamichi.

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The authors declare that they have no conflict of interest.

Electronic supplementary material

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Supplementary material 1 (DOCX 77 kb)

535_2018_1489_MOESM2_ESM.jpg

Supplementary Figure 1: Localization of Over-accumulation of β-catenin. a) β-catenin in duodenal and colorectal adenomas was similarly over-expressed in both the cytoplasm (80.0% vs 86.7%) and nucleus (15.0% vs 20.0%). b) Representative cases of duodenal adenoma. A case with both cytoplasmic and nuclear over-expression of β-catenin (left). A case with only cytoplasmic over-expression of β-catenin with no nuclear staining (right). c) Representative cases of colorectal adenoma. A case with both cytoplasmic and nuclear over-expression of β-catenin (left). A case with only cytoplasmic over-expression of β-catenin with no nuclear staining (right) (JPEG 4840 kb)

535_2018_1489_MOESM3_ESM.jpg

Supplementary Figure 2: a) GSEA demonstrates that FAP-related tumors demonstrate enrichment in the Wnt/β-catenin pathway, with enrichment scores similar to sporadic duodenal tumors. b) The gene expression values of the top up-regulated genes in sporadic duodenal tumors. Only minimal changes can be seen in the gene expression values of these genes in FAP-related duodenal adenomas. c) The gene expression values of the top down-regulated genes in sporadic duodenal tumors. Similarly, only minimal changes can be seen in the gene expression values of these genes in FAP-related duodenal adenomas (JPEG 3758 kb)

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Sakaguchi, Y., Yamamichi, N., Tomida, S. et al. Identification of marker genes and pathways specific to precancerous duodenal adenomas and early stage adenocarcinomas. J Gastroenterol 54, 131–140 (2019). https://doi.org/10.1007/s00535-018-1489-4

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  • DOI: https://doi.org/10.1007/s00535-018-1489-4

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