Differential gene expression in colon carcinoma cells and tissues detected with a cDNA array.
Expression of selected genes coding for proteins with defined cellular functions was analysed in human cell lines derived from normal colonic mucosa, non-mucinous colonic carcinomas and mucinous colonic carcinomas. Altered expression of 10 genes in colon carcinoma cells was found by using a cDNA array; 6 of these alterations (60%) were confirmed by Northern blotting or semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). Among these 6 genes, 3 transcription factors as well as the topoisomerase II alpha and the mitosis inhibitor WEE1Hu gene were significantly suppressed in the tumour cell lines. In addition, the gene coding for the cell cycle inhibitor p21 was overexpressed only in cell lines derived from mucinous carcinomas. The significant suppression of the kinase WEE1Hu gene in carcinoma cells of both phenotypes and the tendency of the mucinous phenotype to overexpress p21 protein were confirmed in human colon carcinoma tissues. Our data show that the cDNA array method permits a correct identification of changes in gene expression with a relatively high accuracy. The different expression of the p21 gene in the non-mucinous and mucinous carcinoma cells supports the hypothesis that these phenotypes may develop along different genetic pathways. The detection of WEE1Hu gene suppression in colon carcinoma cells and tissues suggests its potential role in tumourigenesis.
Backert
S
S
Department of Gastroenterology, Klinikum Benjamin Franklin, Freie Universität, Berlin, Germany.
Gelos
M
M
Kobalz
U
U
Hanski
M L
ML
Böhm
C
C
Mann
B
B
Lövin
N
N
Gratchev
A
A
Mansmann
U
U
Moyer
M P
MP
Riecken
E O
EO
Hanski
C
C
D002288:Adenocarcinoma, Mucinous / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D017209:Apoptosis; D002453:Cell Cycle; D018797:Cell Cycle Proteins; D002460:Cell Line; D003106:Colon; D003110:Colonic Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins / Q000235:genetics; D004260:DNA Repair; D004264:DNA Topoisomerases, Type I / Q000235:genetics; D004250:DNA Topoisomerases, Type II / Q000235:genetics; D018076:DNA, Complementary; D005786:Gene Expression Regulation; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism; D009687:Nuclear Proteins / Q000235:genetics; D011505:Protein-Tyrosine Kinases / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013489:Suppression, Genetic; D014157:Transcription Factors / Q000235:genetics; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
C493956:CDKN1A protein, human; D018797:Cell Cycle Proteins; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D018076:DNA, Complementary; D009687:Nuclear Proteins; D014157:Transcription Factors; D011505:Protein-Tyrosine Kinases; C499349:WEE1 protein, human; D004264:DNA Topoisomerases, Type I; D004250:DNA Topoisomerases, Type II