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Journal of Gastroenterology

, Volume 54, Issue 2, pp 131–140 | Cite as

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

  • Yoshiki Sakaguchi
  • Nobutake YamamichiEmail author
  • Shuta Tomida
  • Chihiro Takeuchi
  • Natsuko Kageyama-Yahara
  • Yu Takahashi
  • Kazuya Shiogama
  • Ken-ichi Inada
  • Masao Ichinose
  • Mitsuhiro Fujishiro
  • Kazuhiko Koike
Original Article—Alimentary Tract

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.

Keywords

Duodenal adenoma Duodenal adenocarcinoma Comprehensive gene expression analysis Wnt/β-catenin pathway Gene set enrichment analysis 

Notes

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.

Author 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.

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.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

535_2018_1489_MOESM1_ESM.docx (78 kb)
Supplementary material 1 (DOCX 77 kb)
535_2018_1489_MOESM2_ESM.jpg (4.7 mb)
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 (3.7 mb)
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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Copyright information

© Japanese Society of Gastroenterology 2018

Authors and Affiliations

  • Yoshiki Sakaguchi
    • 1
  • Nobutake Yamamichi
    • 1
    Email author
  • Shuta Tomida
    • 2
  • Chihiro Takeuchi
    • 1
  • Natsuko Kageyama-Yahara
    • 1
  • Yu Takahashi
    • 1
  • Kazuya Shiogama
    • 3
  • Ken-ichi Inada
    • 3
  • Masao Ichinose
    • 4
  • Mitsuhiro Fujishiro
    • 1
  • Kazuhiko Koike
    • 1
  1. 1.Department of Gastroenterology, Graduate School of MedicineThe University of TokyoTokyoJapan
  2. 2.Graduate School of Medicine, Dentistry and Pharmaceutical SciencesOkayama UniversityOkayamaJapan
  3. 3.Department of PathologyFujita Health University School of MedicineToyoakeJapan
  4. 4.Second Department of Internal MedicineWakayama Medical UniversityWakayamaJapan

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