A profile of differentially expressed genes in primary colorectal cancer using suppression subtractive hybridization.
As a step towards understanding the complex differences between normal cells and cancer cells, we have used suppression subtractive hybridization (SSH) to generate a profile of genes overexpressed in primary colorectal cancer (CRC). From a 35¿ omitted¿000 clone SSH-cDNA repertoire, we have screened 400 random clones by reverse Northern blotting, of which 45 clones were scored as overexpressed in tumor compared to matched normal mucosa. Sequencing showed 37 different genes and of these, 16 genes corresponded to known genes in the public databases. Twelve genes, including Smad5 and Fls353, have previously been shown to be overexpressed in CRC. A series of known genes which have not previously been reported to be overexpressed in cancer were also recovered: Hsc70, PBEF, ribophorin II and Ese-3B. The remaining 21 genes have as yet no functional annotation. These results show that SSH in conjunction with high throughput screening provides a very efficient means to produce a broad profile of genes differentially expressed in cancer. Some of the genes identified may provide novel points of therapeutic intervention.
Hufton
S E
SE
Research Institute Growth and Development, Department of Pathology, University Hospital Maastricht, P.O. Box 5800, 6202 AZ, Maastricht, The Netherlands.
Moerkerk
P T
PT
Brandwijk
R
R
de Bruïne
A P
AP
Arends
J W
JW
Hoogenboom
H R
HR
D000230:Adenocarcinoma / Q000235:genetics* / Q000378:metabolism*; D015152:Blotting, Northern; D003001:Cloning, Molecular; D016207:Cytokines / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic; D015723:Gene Library; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics; D006801:Humans; D008297:Male; D008565:Membrane Proteins / Q000235:genetics; D008875:Middle Aged; D054409:Nicotinamide Phosphoribosyltransferase; D009693:Nucleic Acid Hybridization; D012004:Rectal Neoplasms / Q000235:genetics* / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors / Q000096:biosynthesis* / Q000235:genetics*
D016207:Cytokines; C403306:EHF protein, human; D018840:HSP70 Heat-Shock Proteins; D008565:Membrane Proteins; D014157:Transcription Factors; C034241:ribophorin; D054409:Nicotinamide Phosphoribosyltransferase; C085455:nicotinamide phosphoribosyltransferase, human