Genomic PCR-SSCP analysis of the metastasis associated NM23-H1 (NME1) gene: a study on colorectal cancer.
To facilitate further mutational analysis of NM13-H1, a human metastasis suppressor gene, we have established its genomic organization. NM23-H1 is composed of five exons, spanning a genomic DNA fragment of 10 kb. Using oligonucleotide primers flanking each exon, PCR-SSCP analysis was performed on genomic DNAs of healthy individuals. A common polymorphism, a C to T transition, was detected 30 nucleotides upstream from the 5' splice site flanking exon 1. As NM23-H1 allele loss and altered expression have been reported in colorectal cancer, genomic DNAs of 20 colorectal tumors were analyzed for the presence of gene-specific mutations by PCR-SSCP: no abnormal sequences were detected within the coding and splice site regions of the NM23-H1 gene. This finding suggests that NM23-H1 mutations are rare events in human colorectal cancer.
Bafico
A
A
Istituto Nazionale per la Ricerca sul Cancro, Genova, Italy.
Varesco
L
L
De Benedetti
L
L
Caligo
M A
MA
Gismondi
V
V
Sciallero
S
S
Aste
H
H
Ferrara
G B
GB
Bevilacqua
G
G
D001483:Base Sequence; D002886:Chromosomes, Human, Pair 17; D003001:Cloning, Molecular; D015179:Colorectal Neoplasms / Q000235:genetics* / Q000473:pathology; D003360:Cosmids; D004252:DNA Mutational Analysis; D017931:DNA Primers; D018076:DNA, Complementary / Q000032:analysis; D005091:Exons; D015723:Gene Library; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D009362:Neoplasm Metastasis / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods; D011110:Polymorphism, Genetic; D012326:RNA Splicing; D015183:Restriction Mapping
D017931:DNA Primers; D018076:DNA, Complementary