Differential expression of mucins and trefoil peptides in native epithelium, Barrett's metaplasia and squamous cell carcinoma of the oesophagus.

BACKGROUND AND AIMS: In humans, trefoil peptides (TFF peptides) and some mucins have been reported to be expressed in a cell-specific manner at mucosal surfaces of normal gastrointestinal tissues. Neoplastic conditions cause characteristic changes of these expression patterns. To study such patterns in Barrett's metaplasia and squamous cell carcinoma of the oesophagus (SCC), the distribution of MUC1, MUC2, MUC5AC and the three TFF peptides (TFF1, TFF2 and TFF3) was investigated. METHODS: In 40 archival samples of SCC and in 21 samples of Barret's metaplasia, expression of the three mucins and two TFF peptides (TFF1 and TFF2) was assessed by specific antibodies. Reverse transcriptase/polymerase chain reaction amplification (RT-PCR) was performed on frozen tissue samples from the 11 biopsies of SCC for the three TFF peptides. RESULTS: Immunohistochemical tests for MUC2 and TFF2 were negative both in samples of Barret's metaplasia and in SCC. MUC1 expression was detected in 57.5% of the tumour samples, while TFF1 and MUC5AC were found in 10% and 7.5% of the cases respectively. In Barrett's metaplasia MUC1 was detected in 90.5% of the cases and TFF1 and MUC5AC in all of them. RT-PCR analysis revealed a more complex pattern: TFF1 and TFF3 expressed the corresponding mRNA in all samples investigated; the third member, TFF2, was active in 45.5% of the carcinoma biopsies and not in the corresponding native tissue. CONCLUSIONS: This finding in oesophageal carcinoma contrasts with the situation found in normal and neoplastic stomach epithelium where TFF1 and TFF2 are found co-expressed and TFF3 remains silent. Interestingly, MUC1 is expressed in a significant proportion of SCC. Both in Barett's metaplasia and in SCC the expression of MUC5AC mirrors the TFF1 synthesis in intensity and spatial distribution.

Expression pattern of gastrointestinal markers in native colorectal epithelium, lesions, and carcinomas.

Three small peptides with a typical cysteine-rich domain (TFF or P-domain) display a specific folding structure (trefoil); they are abundantly expressed on mucosal surfaces of normal and neoplastic gastrointestinal tissues. This epithelial location coincides with mucin secretion and, although not proven beyond doubt, this association is suggestive of their function in maintenance of surface integrity. Using normal colon epithelium, premalignant lesions and tumor samples and specific antibodies we studied expression of these peptides in colorectal carcinomas. RT PCR was performed to extend the sensitivity of the assays. While coexpression of pS2, hITF, MUC1 and MUC2 was demonstrated, MUC5 was absent and no simultaneous activity of pS2 and SML1 (as in gastric mucosa and carcinoma) was noted in rectal tumors. Actively transcribed and expressed cytokeratin 20 and GAPDH were used as experimental controls for immunostaining and RT-PCR, respectively.

Human lymphocyte culture: intrinsic factors influencing the quality of chromosome preparations.

In a cytogenetic study of the consecutive cell cycles (M1, M2 etc.) from human lymphocyte cultures after 5-bromodeoxyuridine incorporation followed by differential Giemsa staining, the metaphases of M2 and subsequent in vitro divisions have been found to spread better than those of M1 cells, as if the cells of in vitro generation suffer some membrane alterations. Moreover presence of S or G2 phase, leukocytes in a blood sample may result into early proliferation in a PHA stimulated culture system, so that the metaphase yield at 72 h will be larger than otherwise, a sizeable portion of which, being in M2 or subsequent cell cycles, will spread better. Thus such individual or transient intrinsic factors of haematologic state may influence the success of a leukocyte culture and the quality of chromosome preparation.