Rapid detection of ras oncogenes in human tumors: applications to colon, esophageal, and gastric cancer.
We have developed a rapid, nonradioactive large scale method for the detection of ras oncogenes in human tumors. DNA is amplified by the polymerase chain reaction (PCR), and then digested with specific restriction enzymes to detect either endogenous or primer-mediated Restriction Fragment Length Polymorphisms (RFLPs). We report here that three of 15 colon tumors tested contain K-ras codon 12 aspartic acid mutations and one, along with the HCT 116 colon carcinoma cell line, contains a K-ras codon 13 aspartic acid mutation. On the other hand, we did not detect H- or K-ras codon 12 mutations or the K-ras codon 13 aspartic acid mutation in 25 esophageal and 27 gastric cardia tumors isolated from patients in Lin-xing County, China. By incorporating nucleotide substitutions in PCR primers, this method can be applied towards the rapid, non-radioactive screening of virtually any genetic disease caused by known point mutations.
Jiang
W
W
Comprehensive Cancer Center, College of Physicians and Surgeons, Columbia University, New York, New York 10032.
Kahn
S M
SM
Guillem
J G
JG
Lu
S H
SH
Weinstein
I B
IB
D003062:Codon; D003110:Colonic Neoplasms / Q000235:genetics*; D004938:Esophageal Neoplasms / Q000235:genetics*; D005784:Gene Amplification; D011905:Genes, ras; D006801:Humans; D009154:Mutation; D013274:Stomach Neoplasms / Q000235:genetics*; D014407:Tumor Cells, Cultured
D003062:Codon