Detection of K-ras mutations by denaturing gradient gel electrophoresis (DGGE): a study on pancreatic cancer.
In pancreatic neoplasias mutations in the first exon (codon 12) of K-ras gene occur at high frequency and seem to have a diagnostic significance. We set up the DGGE conditions to search for these mutations in pancreatic tumor sample DNAs. All samples were directly classified by simply comparing their DGGE patterns with those of control cell lines carrying known K-ras base substitutions. We found a mutation frequency of 73% in pancreatic adenocarcinoma, whereas no mutations were observed in benign lesions. The non-isotopic method we used turned out to be rapid and sensitive. DGGE could therefore be utilized for the detection of K-ras mutations in pancreatic lesions, to evaluate their actual or potential malignancy. In general, DGGE could be useful for K-ras gene screening on pathological tissue samples.
Pellegata
N S
NS
Department of Genetics and Microbiology, Pavia University, Italy.
Losekoot
M
M
Fodde
R
R
Pugliese
V
V
Saccomanno
S
S
Renault
B
B
Bernini
L F
LF
Ranzani
G N
GN
D000230:Adenocarcinoma / Q000235:genetics* / Q000601:surgery; D001483:Base Sequence; D002460:Cell Line; D003062:Codon; D003537:Cystadenoma / Q000235:genetics* / Q000601:surgery; D004273:DNA, Neoplasm / Q000235:genetics / Q000302:isolation & purification; D011905:Genes, ras; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D009838:Oligodeoxyribonucleotides; D010182:Pancreatic Diseases / Q000235:genetics / Q000473:pathology; D010190:Pancreatic Neoplasms / Q000235:genetics* / Q000601:surgery; D016133:Polymerase Chain Reaction; D014407:Tumor Cells, Cultured
D003062:Codon; D004273:DNA, Neoplasm; D009838:Oligodeoxyribonucleotides