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. 1999 Nov;45(5):686–692. doi: 10.1136/gut.45.5.686

Villous, hypermucinous mucosa in long standing ulcerative colitis shows high frequency of K-ras mutations

S Andersen 1, T Lovig 1, O Clausen 1, A Bakka 1, O Fausa 1, T Rognum 1
PMCID: PMC1727707  PMID: 10517904

Abstract

BACKGROUND—K-ras mutation is one of the first genetic alterations in classical colorectal carcinogenesis.
AIMS—To investigate the role of K-ras mutations in carcinogenesis, in long standing ulcerative colitis.
METHODS—A total of 161 microdissected and 100 DNA samples from 13 patients were analysed for K-ras codons 12 and 13 mutations by means of a combination of enriched polymerase chain reaction amplification and temporal temperature gradient electrophoresis.
RESULTS—K-ras mutations were found in 21/161 (13%) microdissected samples in 7/13 large bowels (16 and five in codons 12 and 13, respectively), and in 10/100 (10%) mucosal DNA samples (six and four, respectively). One of four patients with six adenocarcinomas had a K-ras mutation in a carcinoma, as well as one of two patients with large dysplasia associated lesion or mass (DALM). Eight of 13 (61%) areas with villous architecture and large, distended goblet cells, had a K-ras mutation, which was significantly more frequent than in low grade dysplasia (one of 23, 4%) but did not reach significance versus high grade dysplasia (four of 14, 28.5%). K-ras mutations were found in one of 20 (5%) flat lesions indefinite for dysplasia, two of 14 (14%) in non-villous, hypermucinous mucosa, and in one of 57 flat areas negative for dysplasia.
CONCLUSION—The highest K-ras mutation frequency was found in villous, hypermucinous mucosa. We suggest that this entity should be investigated further as a potential risk lesion for cancer development. It may represent a pathway directly from non-classical dysplasia to cancer, not previously described.


Keywords: K-ras mutations; ulcerative colitis; dysplasia; dysplasia associated lesion or mass

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Figure 1  .

Figure 1  

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Section from large bowel mucosa of patient 2, rich in goblet cells, but without dysplasia, (A) before and (B) after microdissection.

Figure 2  .

Figure 2  

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Mucosal specimen from patient 11, with villous growth pattern and elongated, distended goblet cells without cytological atypia. This area was K-ras mutation positive.

Figure 3  .

Figure 3  

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TTGE gel showing migration patterns of K-ras exon 1 fragments. All lanes show different migration patterns at the upper bands (heteroduplex bands), indicating different K-ras mutations. The lower band is the homoduplex band. Lanes 1-5 show codon 12 mutations: (1) GTT (valine); (2) TGT (cysteine); (3) GAT (aspartate); (4) GCT (alanine); and (5) AGT (serine). Lane 6 shows a codon 13 GAC mutation (aspartate).

Figure 4  .

Figure 4  

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(A) Automated sequence analysis of K-ras, showing wild type sequence GGT in codon 12. This is an inverse analysis, and should be read from right to left. (B). K-ras mutation in the second base in codon 12, giving the sequence GAT (aspartate) instead of GGT (glycine).

Selected References

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