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In situ detection of unexpected patterns of mutant p53 gene expression in non-small cell lung cancers
- Source :
- Oncogene. 20(20)
- Publication Year :
- 2000
-
Abstract
- Many solid tumors, including non-small cell lung cancers (NSCLCs), are characterized by heterogenous expression of p53 protein in the neoplastic cells. To analyse the molecular implications of this finding, we examined topographic distribution of p53 mutations using in situ polymerase chain reaction (PCR) in primary NSCLCs, showing distinct patterns of variable p53 overexpression by immunohistochemistry. Unique sets of primers for each mutation were designed, and optimal PCR conditions were determined by standard PCR using DNA from cloned mutants or cell lines established from these tumors. All tumor cell nuclei, regardless of the status of p53 overexpression, demonstrated homogeneous distribution of mutant p53 with specific primers, indicating that only subgroups of the mutated cells overexpressed p53 protein. In situ reverse transcription (RT)-PCR was applied to detect mutant mRNA in the individual tumor cells using specific primers. We found that in each case the distribution of mutant p53 mRNA coincided with that of immunohistochemical overexpression of p53 protein. Our results suggest that the regulation of mutant p53 expression, but not the genotype, is heterogeneous in the neoplastic cells. The topographic genomapping of p53 in NSCLC using in situ PCR provides a novel approach to view molecular mechanisms of lung carcinogenesis.
- Subjects :
- Cancer Research
Lung Neoplasms
Mutant
Biology
medicine.disease_cause
law.invention
law
Carcinoma, Non-Small-Cell Lung
Gene expression
Genetics
medicine
Humans
Point Mutation
Molecular Biology
Gene
Polymerase chain reaction
Aged
Regulation of gene expression
Mutation
Reverse Transcriptase Polymerase Chain Reaction
Middle Aged
Genes, p53
Molecular biology
Immunohistochemistry
Reverse transcriptase
Gene Expression Regulation, Neoplastic
Tumor Suppressor Protein p53
Carcinogenesis
Subjects
Details
- ISSN :
- 09509232
- Volume :
- 20
- Issue :
- 20
- Database :
- OpenAIRE
- Journal :
- Oncogene
- Accession number :
- edsair.doi.dedup.....f6085c87c594de9d5358ce619153963e