Radiolabeled polymerase chain reaction assay for detection of ras oncogene point mutations in tumors.
The human ras gene plays a fundamental role in the transduction of extracellular signals to the nucleus, thereby regulating cell growth and differentiation. Point mutations in the ras gene convert it into a transforming oncogene that has been found in many solid and hematologic malignancies. We describe a rapid and sensitive assay based on a radiolabeled polymerase chain reaction followed by restriction enzyme digestion that we have adapted for differentiating between the wild-type and mutant ras genes. This assay should prove useful in the analysis of ras gene point mutations in clinical tumor specimens in which ras oncogene activation is an early event in carcinogenesis.
Aoki
Y
Y
Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown 02129.
Lee
J C
JC
Pillai
S
S
Isselbacher
K J
KJ
Rustgi
A K
AK
D001483:Base Sequence; D015139:Blotting, Southern; D003062:Codon; D003110:Colonic Neoplasms / Q000235:genetics*; D004273:DNA, Neoplasm / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D015252:Deoxyribonucleases, Type II Site-Specific; D011905:Genes, ras; D006801:Humans; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D010761:Phosphorus Radioisotopes; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D014407:Tumor Cells, Cultured
D003062:Codon; D004273:DNA, Neoplasm; D010761:Phosphorus Radioisotopes; C046545:CCWGG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific