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 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.
Furuwatari
C
C
Central Clinical Laboratories, Shinshu University Hospital, Matsumoto, Japan.
Yagi
A
A
Yamagami
O
O
Ishikawa
M
M
Hidaka
E
E
Ueno
I
I
Furihata
K
K
Ogiso
Y
Y
Katsuyama
T
T
D002460:Cell Line; D004252:DNA Mutational Analysis; D004273:DNA, Neoplasm / Q000032:analysis*; D005069:Evaluation Studies as Topic; D005189:False Positive Reactions; D016158:Genes, p53 / Q000235:genetics*; D006801:Humans; D007124:Immunoenzyme Techniques; D017404:In Situ Hybridization, Fluorescence; D019656:Loss of Heterozygosity; D009154:Mutation; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D017422:Sequence Analysis, DNA; D016159:Tumor Suppressor Protein p53 / Q000235:genetics* / Q000378:metabolism
D004273:DNA, Neoplasm; D016159:Tumor Suppressor Protein p53