RNA sequencing of sessile serrated colon polyps identifies differentially expressed genes and immunohistochemical markers

PLoS One. 2014 Feb 12;9(2):e88367. doi: 10.1371/journal.pone.0088367. eCollection 2014.

Abstract

Background: Sessile serrated adenomas/polyps (SSA/Ps) may account for 20-30% of colon cancers. Although large SSA/Ps are generally recognized phenotypically, small (<1 cm) or dysplastic SSA/Ps are difficult to differentiate from hyperplastic or small adenomatous polyps by endoscopy and histopathology. Our aim was to define the comprehensive gene expression phenotype of SSA/Ps to better define this cancer precursor.

Results: RNA sequencing was performed on 5' capped RNA from seven SSA/Ps collected from patients with the serrated polyposis syndrome (SPS) versus eight controls. Highly expressed genes were analyzed by qPCR in additional SSA/Ps, adenomas and controls. The cellular localization and level of gene products were examined by immunohistochemistry in syndromic and sporadic SSA/Ps, adenomatous and hyperplastic polyps and controls. We identified 1,294 differentially expressed annotated genes, with 106 increased ≥10-fold, in SSA/Ps compared to controls. Comparing these genes with an array dataset for adenomatous polyps identified 30 protein coding genes uniquely expressed ≥10-fold in SSA/Ps. Biological pathways altered in SSA/Ps included mucosal integrity, cell adhesion, and cell development. Marked increased expression of MUC17, the cell junction protein genes VSIG1 and GJB5, and the antiapoptotic gene REG4 were found in SSA/Ps, relative to controls and adenomas, were verified by qPCR analysis of additional SSA/Ps (n = 21) and adenomas (n = 10). Immunohistochemical staining of syndromic (n≥11) and sporadic SSA/Ps (n≥17), adenomatous (n≥13) and hyperplastic (n≥10) polyps plus controls (n≥16) identified unique expression patterns for VSIG1 and MUC17 in SSA/Ps.

Conclusion: A subset of genes and pathways are uniquely increased in SSA/Ps, compared to adenomatous polyps, thus supporting the concept that cancer develops by different pathways in these phenotypically distinct polyps with markedly different gene expression profiles. Immunostaining for a subset of these genes differentiates both syndromic and sporadic SSA/Ps from adenomatous and hyperplastic polyps.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenoma / genetics*
  • Adenoma / metabolism
  • Adenoma / pathology*
  • Aged
  • Antigens, Neoplasm / metabolism
  • Cluster Analysis
  • Colon / metabolism
  • Colonic Neoplasms / genetics
  • Colonic Neoplasms / metabolism
  • Colonic Polyps / genetics*
  • Colonic Polyps / pathology*
  • Colonoscopy
  • Connexins / metabolism
  • DNA Mutational Analysis
  • Female
  • Gene Expression Profiling
  • Gene Expression Regulation
  • Humans
  • Immunohistochemistry
  • Lectins, C-Type / metabolism
  • Male
  • Membrane Glycoproteins / metabolism
  • Middle Aged
  • Mucins / metabolism
  • Pancreatitis-Associated Proteins
  • Polymerase Chain Reaction
  • Proto-Oncogene Proteins B-raf / genetics
  • Sequence Analysis, RNA / methods*

Substances

  • Antigens, Neoplasm
  • Connexins
  • GJB5 protein, human
  • Lectins, C-Type
  • MUC17 protein, human
  • Membrane Glycoproteins
  • Mucins
  • Pancreatitis-Associated Proteins
  • REG4 protein, human
  • VSIG1 protein, human
  • BRAF protein, human
  • Proto-Oncogene Proteins B-raf