title: Frequency of allele loss of DCC, p53, RBI, WT1, NF1, NM23 and APC/MCC in colorectal cancer assayed by fluorescent multiplex polymerase chain reaction.
abstract: We report here the use of multiplex fluorescent polymerase chain reaction (PCR) for quantitative allele loss detection using microsatellites with 2-5 base pair repeat motifs. Allele loss of APC, DCC, p53 and RB1 in colorectal tumours has been reported previously using a variety of methods. However, not all workers used intragenic markers. We have used microsatellite polymorphisms which map within, or are closely linked to, these tumour-suppressor gene loci in order to determine whether these loci are indeed the targets for alteration in colorectal cancer. In addition, we have assayed two other tumour-suppressor genes, WT1 and NF1, to see whether they play a role in colorectal carcinogenesis. The putative metastasis-suppressor gene, NM23, was also investigated since there have been conflicting reports about its involvement in colorectal carcinogenesis. Allele loss was detected at the DCC (29%), p53 (66%), RB1 (50%) and NF1 (14%) loci and in the APC/MCC region (50%), but not at the WT1 or NM23 loci. These rapid, and mostly gene-specific, fluorescent multiplex PCR assays for allele loss detection could be modified to devise a single molecular diagnostic test for the important lesions in colorectal cancer.
authors.affiliation: Centre for Cancer Studies, Research School of Medicine, University of Leeds, UK.
authors.forename: L
authors.initials: L
authors.lastname: Cawkwell
authors.forename: F A
authors.initials: FA
authors.lastname: Lewis
authors.forename: P
authors.initials: P
authors.lastname: Quirke
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics*; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000483:Alleles; D001483:Base Sequence; D015179:Colorectal Neoplasms / Q000235:genetics*; D004276:DNA, Satellite / Q000032:analysis / Q000235:genetics; D005260:Female; D005453:Fluorescence; D017353:Gene Deletion; D017491:Genes, APC / Q000235:genetics; D016163:Genes, DCC / Q000235:genetics; D017431:Genes, MCC / Q000235:genetics; D016514:Genes, Neurofibromatosis 1 / Q000235:genetics; D016161:Genes, Retinoblastoma / Q000235:genetics; D016147:Genes, Tumor Suppressor; D016162:Genes, Wilms Tumor; D016158:Genes, p53 / Q000235:genetics; D005819:Genetic Markers; D006801:Humans; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D020559:Monomeric GTP-Binding Proteins; D054778:NM23 Nucleoside Diphosphate Kinases; D009701:Nucleoside-Diphosphate Kinase; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic; D012680:Sensitivity and Specificity; D014157:Transcription Factors / Q000235:genetics
chemical_list: D004276:DNA, Satellite; D005819:Genetic Markers; D054778:NM23 Nucleoside Diphosphate Kinases; D014157:Transcription Factors; C517938:NME1 protein, human; D009701:Nucleoside-Diphosphate Kinase; D020559:Monomeric GTP-Binding Proteins