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. 2000 Sep;47(3):349–356. doi: 10.1136/gut.47.3.349

Role of fucosyltransferases in the association between apomucin and Lewis antigen expression in normal and malignant gastric epithelium

A Lopez-Ferrer 1, C de Bolos 1, C Barranco 1, M Garrido 1, J Isern 1, I Carlstedt 1, C Reis 1, J Torrado 1, F Real 1
PMCID: PMC1728024  PMID: 10940270

Abstract

BACKGROUND—In normal gastric epithelium, MUC5AC is detected in superficial epithelium associated with Lewis type 1 antigens and MUC6 is detected in antral glands with Lewis type 2. Therefore, the stomach constitutes an excellent model to examine the role of glycosyltransferases in determining the specificity of apomucin glycosylation.
AIMS—To determine the molecular basis of this association and to examine changes in expression of gastric and intestinal apomucins and their association with Lewis antigens during the gastric carcinogenesis process.
METHODS—Fucosyltransferase (FUT1, FUT2, FUT3) and mucin (MUC5AC, MUC6) transcripts were detected using reverse transcription-polymerase chain reaction. Apomucin (MUC2, MUC4, MUC5AC, MUC6) and Lewis antigen (types 1 and 2) expression were analysed using single and double immunohistochemistry and in situ hybridisation.
RESULTS—In the normal stomach, FUT1 is exclusively detected associated with MUC6; FUT2 is only detected when MUC5AC is present. This co-regulation is lost in gastric tumours, as is differential expression of MUC5AC and MUC6 in normal gastric epithelial cells. In gastric tumours, especially those with the intestinal phenotype, MUC2 and MUC4 genes are upregulated, and gastric-type and intestinal-type mucins are coexpressed. These changes are early events in the gastric carcinogenesis process, as they are detected in intestinal metaplasia.
CONCLUSIONS—The glycosylation pattern found in normal gastric epithelium is dictated by the specific set of fucosyltranferases expressed by the cells rather than by the apomucin sequence. The development of intestinal metaplasia and gastric cancer is associated with the appearance of cellular phenotypes that are absent from normal epithelium.


Keywords: fucosyltransferases; gastric carcinogenesis; gastric mucins; Lewis antigens

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Figure 1  .

Figure 1  

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Mucin and fucosyltransferase mRNA expression in gastric mucosa scrapings detected by RT-PCR (S, superficial epithelium; G, deep glands). MUC6 and FUT1 were detected only in the deep gland scrapings. MUC5AC and FUT2 were detected in both superficial epithelium and deep glands, suggesting that cells from the superficial epithelium contaminated the deep gland fraction.

Figure 2  .

Figure 2  

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Mucin expression in gastric tumours. (A) MUC4 detected by immunohistochemistry in an intestinal-type tumour. (B) MUC4 expression by in situ hybridisation in a diffuse-type tumour (original magnification: A ×200; B ×100). (C) Coexpression of several apomucin genes in gastric tumours: western blot detection of MUC2, MUC4, MUC5AC, and MUC6 apomucins in lysates of intestinal (i) (1, 2, 4) and diffuse (d) (3) gastric tumours. Nitrocellulose membranes were treated with 100 mM sodium periodate before immunodetection. Several apomucins are detected in the same tumour sample. Markers (−) indicate the borders between running and stacking gels.

Figure 3  .

Figure 3  

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Gastric cancers contain cells displaying abnormal patterns of apomucin/Lewis antigen coexpression. (A) Double labelling immunohistochemical detection of MUC5AC (blue) and MUC6 expression (light brown) cells. Doubly labelled cells display a dark brown colour (original magnification ×400). (B) Double labelling immunoelectron microscopy using antibodies against MUC5AC (CLH2) and MUC6 (anti-MUC6.1) and sections from Lowicryl 4KM embedded tissues. Mouse antibodies were detected using 15 nm gold particles (arrowheads) and rabbit antibodies using 5 nm gold particles (arrows) (original magnification ×63 000). (C) Double labelling immunohistochemical detection of MUC5AC (brown) and Lewis y (blue). (D) Double labelling immunohistochemical detection of MUC6 (brown) and Lewis b (blue). Arrowheads in (A), (C), and (D) indicate coexpression in tumour cells (original magnification ×400).

Figure 4  .

Figure 4  

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RT-PCR analysis of fucosyltransferase (FUT1 and FUT2) and mucin (MUC5AC and MUC6) transcripts in two gastric tumours (A and B). There was no association between the presence of MUC5AC and MUC6, and FUT2 and FUT1 transcripts, respectively.

Figure 5  .

Figure 5  

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Expression of apomucins and Lewis antigens in intestinal metaplasia. Double labelling immunofluorescence detection of: (A) MUC2 (green) and MUC6 (red); (B) MUC4 (green) and Lewis b (red); (C) MUC5AC (green) and Lewis y (red). In incomplete intestinal metaplasia, coexpression of gastric and intestinal apomucins at the single cell level is demonstrated. In addition, the association between apomucin and Lewis antigens present in normal stomach is lost (original magnification: A ×630; B, C ×400).

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