A Comprehensive System to Explorep53Mutations

To establish an effective and reliable system for the detection of p53 mutations, we evaluated the detection efficiencies of nonisotopic polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP), fluorescence in situ hybridization (FISH), and immunohistochemistry. Ten cell lines (AsPc1, BxPc3, Miapaca2, Panc1, Colo320-011, Lovo, MCF7, LNCaP, HL-60, and Daudi), a peripheral blood sample from a patient with a p53 germline mutation (p53GML), and a normal peripheral blood sample were used for examination. Direct nucleotide sequencing identified p53 mutations in 7 of 12 samples (AsPc1, BxPc3, Miapaca2, Panc1, Colo320-011, HL-60, and p53GML). The nonisotopic PCR-SSCP detected anomalies of the PCR fragments in 5 cell lines. In the FISH analysis, 2 cell lines exhibited loss of heterozygosity of the p53 locus. Immunohistochemistry detected an accumulation of the abnormal p53 in 4 cell lines. The combination of these 3 methods produced no false-negative or false-positive results. This combination may be an excellent and beneficial system for the clinical diagnosis of the various human cancers.