Crude isolation of DNA from unselected human pancreatic tissue and amplification by the polymerase chain reaction of Ki-ras oncogene to detect point mutations in pancreatic cancer.
Human pancreatic tissue obtained during surgery was divided into two samples, one stored in formalin for histology and the other frozen directly for DNA studies. A small bit of the frozen tissue was excised without prior differentiation of malignant from non-malignant tissue and crude DNA prepared. The DNA was used to amplify the Ki-ras oncogenes with the polymerase chain reaction (PCR) using primers for codons 12, 13 and 61. The PCR products were hybridized with oligonucleotides to detect mutations in these codons. No mutations were seen in patients histologically free from pancreatic cancer, having cancer of the papilla of Vater or endocrine pancreatic cancer. In the 9 cases of pancreatic cancer, point mutations were detected in codon 12 in 7 cases and in codon 13 in one patient. Thus, the PCR technique can be used to detect point mutations in Ki-ras oncogenes even in crude DNA without selection of malignant from non-malignant tissue.
Masson
P
P
Department of Clinical Chemistry, University Hospital, Lund, Sweden.
Andrén-Sandberg
A
A
D003062:Codon / Q000235:genetics*; D004252:DNA Mutational Analysis; D004273:DNA, Neoplasm / Q000302:isolation & purification*; D011905:Genes, ras; D006801:Humans; D009154:Mutation / Q000235:genetics*; D021141:Nucleic Acid Amplification Techniques; D010179:Pancreas / Q000737:chemistry*; D010190:Pancreatic Neoplasms / Q000235:genetics*; D016133:Polymerase Chain Reaction
D003062:Codon; D004273:DNA, Neoplasm