Analysis of expressed N-ras mutations in human melanoma short-term cell lines with allele specific restriction analysis induced by the polymerase chain reaction

摘要: Mutations in ras genes have been found the DNA of numerous cancer types including melanomas, but expression these mutations melanomas has not yet addressed. We used polymerase chain reaction (PCR) and allele-specific restriction analysis (ASRA) to determine frequency expressed N-ras on 25 short-term melanoma tissue culture samples. cDNA generated using reverse transcriptase from whole cells was as PCR template. 14 secondary cultures that varied differentiation patterns were analysed. Only 2 express mutations; both, mutation localised one first two positions 61st codon N-ras. These tumour lines, KMI-M8412a KMI-M8412b, established separate deposits same patient. Codons 12 13 be free all lines studied. 8 primary 3 unclassified skin lesions also analysed mutations. results suggest may play such an important role tumorigenesis is speculated by others.