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Clinical Investigations |
Molecular Diagnostic Laboratory, Department of Clinical Biochemistry, Aarhus University Hospital, DK-8200 Aarhus, Denmark [K. B-D., L. L. C., S. H. O., C. M. F., T. F. Ø.]; Department of Medical Genetics, Haartman Institute, University of Helsinki, FIN-00014 Helsinki, Finland [P. L., L. A. A.]; and Surgical Department L [S. L.] and Institute of Pathology [F. B. S., R. H.], Aarhus University Hospital, DK-8000 Aarhus C, Denmark
Understanding molecular alterations in colorectal cancer (CRC) is needed to define newbiomarkers and treatment targets. We used oligonucleotide microarrays tomonitor gene expression of about 6,800 known genes and 35,000 expressed sequence tags (ESTs) on five pools (four to six samples in each pool) of total RNA from left-sided sporadic colorectal carcinomas. We compared normal tissue to carcinoma tissue from Dukes’ stages A-D (noninvasive to distant metastasis) and identified 908 known genes and 4,155 ESTs that changed remarkably from normal to tumor tissue. Based on intensive filtering 226 known genes and 157 ESTs were found to be highly relevant for CRC. The alteration of known genes was confirmed in >70% of the cases by array analysis of 25 single samples. Two-way hierarchical average linkage cluster analysis clustered normal tissue together with Dukes’ A, clustered Dukes’ B with Dukes’ C, and clustered Dukes’ D separately. Real-time PCR of 10 known genes and 5 ESTs demonstrated excellent reproducibility of the array-based findings. The most frequently altered genes belonged to functional categories of metabolism (22%), transcription and translation (11%), and cellular processes (9%). Fifteen nuclear encoded mitochondrial proteins were all down-regulated in CRC. We identified several chromosomal locations with clusters of either potential oncogenes or potential tumor suppressors. Some of these, such as aminopeptidase N/CD13 and sigma B3 protein on chromosome 15q25, coincided with a high frequency of loss of heterozygosity. The genes and ESTs presented in this study encode new potential tumor markers as well as potential novel therapeutic targets for prevention or therapy of CRC.
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