CDX1 expression is reduced in colorectal carcinoma and is associated with promoter hypermethylation.
The CDX1 homeobox gene encodes a transcription factor specifically expressed in normal intestinal and colonic epithelia, and CDX1 gene expression is affected during colorectal tumour progression. In this study, real-time quantitative RT-PCR was used to investigate CDX1 expression in 26 colorectal carcinomas. Reduced expression of CDX1 was observed in 19 of 26 colon carcinomas compared to matched normal colonic mucosa: the decrease in CDX1 expression ranged between 0.10 and 0.79 (21-90% decrease; mean 64.75% +/-22; p = 0.001). Mutation and loss of heterozygosity (LOH) analyses were then used to determine if reduced CDX1 expression was due to genetic alteration. No CDX1 gene mutations, but two known polymorphisms in exon 1, were observed. LOH was observed in 33% of the tumours investigated but this was not related to CDX1 expression. Since aberrant promoter methylation is a well-known mechanism that participates in gene silencing, the methylation status of the CDX1 5' CpG island promoter was also investigated. PCR amplification of bisulphite-treated DNA followed by cloning was performed in 7 carcinomas that showed low expression of CDX1 and in 1 colonic carcinoma without reduced expression. Promoter hypermethylation occurred in carcinomas in which CDX1 reduced expression was present. These results suggest that CDX1 promoter hypermethylation is one of the molecular mechanisms that accounts for reduced CDX1 gene expression in colorectal carcinoma.
Pilozzi
Emanuela
E
Department of Laboratory Medicine and Pathology, Sant'Andrea Hospital, University La Sapienza, Rome, Italy. emanuela.pilozzi@uniroma1.it
Onelli
Mariadele Rapazzotti
MR
Ziparo
Vincenzo
V
Mercantini
Paolo
P
Ruco
Luigi
L
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001483:Base Sequence; D003110:Colonic Neoplasms / Q000235:genetics; D015179:Colorectal Neoplasms / Q000235:genetics*; D018899:CpG Islands / Q000235:genetics; D004252:DNA Mutational Analysis / Q000379:methods; D004273:DNA, Neoplasm / Q000235:genetics; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics*; D018398:Homeodomain Proteins / Q000235:genetics*; D006801:Humans; D019656:Loss of Heterozygosity / Q000235:genetics; D008297:Male; D008745:Methylation; D008875:Middle Aged; D011110:Polymorphism, Genetic / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods
C491273:CDX1 protein, human; D004273:DNA, Neoplasm; D018398:Homeodomain Proteins