Evaluation of SAT-1, SAT-2 and GalNAcT-1 mRNA in colon cancer by real-time PCR
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By qualitative and quantitative PCR, we evaluated the expression of three messengers coding for SAT-1, SAT-2 and GalNAcT-1 in human samples of intestinal cancer and some cell lines (breast cancer and melanomas). Qualitative PCR demonstrated, in human tissues but not in the cell lines examined, the presence of an mRNA that lacks hexon 3; experiments performed on transfected SKMEL-28 excluded a regulative role of this noncanonical mRNA. Data from real-time PCR, statistically analysed by ANOVA indicated that the mRNA expression of all the considered glycosyltransferases (SAT-1, SAT-2 and GalNAcT-1) was significantly different in tumours versus their own control. The ganglioside patterns in the examined samples did not correlate with mRNA expression; this finding demonstrates that ganglioside expression is the result of a very complex balance between anabolic and catabolic enzyme activities.
Although this study is still preliminary, it opens a new possibility for neoplastic prognosis finding potential molecular markers among the mRNAs that codify for glycosyltransferases.
KeywordsColon cancer Ganglioside Molecular markers Real-Time PCR Tumour Glycosyltransferases
- Ganglioside nomenclature is in accordance with Svennerholm  and the IUPAC-IUBMB recom- mendations .
High Performance Thin Layer Chromatography
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This research was authorized by San Carlo Borromeo Hospital (Milan, Italy) after written informed consent was received from the patients according to the protocol submitted and approved by the local ethical committee. We wish to thank Professor Binelli, DBSM Università dell’Insubria, Varese, for help with the statistical analysis. Supported by a grant from Consorzio Italiano per le Biotecnologie (C.I.B.) 2003.
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