A COMPREHENSIVE SYSTEM TO EXPLORE P53 MUTATIONS

To establish an effective and reliable system for the detection of p53 mutations, we evaluated the detection efficiencies of nonisotopic pol ymerase chain reaction-single-strand conformation polymorphism (PCR-SS CP), fluorescence in situ hybridization (FISH), and immunohistochemist ry. Ten cell lines (AsPc1, BxPc3, Miapaca2, Pane1, Colo320-011, Love, MCF7, LNCaP, HL-60, and Daudi), a peripheral blood sample from a patie nt with a p53 germline mutation (p53GML), and a normal peripheral bloo d sample were used for examination. Direct nucleotide sequencing ident ified p53 mutations in 7 of 12 samples (AsPc1, BxPc3, Miapaca2, Pane1, Colo320-011, HL-60, and p53GML). The nonisotopic PCR-SSCP detected an omalies of the PCR fragments in 5 cell lines. In the FISH analysis, 2 cell lines exhibited loss of heterozygosity of the p53 locus. Immunohi stochemistry detected an accumulation of the abnormal p53 in 4 cell li nes. The combination of these 3 methods produced no false-negative or false-positive results. This combination may be an excellent and benef icial system for the clinical diagnosis of the various human cancers.