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Lack of mutations in K-ras codons 12 and 13 in human atherosclerotic lesions
M Bogliolo1, G Fronza, P Campomenosi
1Center for the Study of Tumors of Environmental Origin, National Institute for Cancer Research (IST), Genoa, Italy.
Abstract:
In the framework of a project investigating the possible involvement of cancer biomarkers in human atherogenesis, we evaluated the occurrence of K-ras mutations in the DNA extracted from smooth muscle cells of abdominal aorta atherosclerotic lesions. The molecular analysis of the DNA from 32 surgical specimens, using PCR-based denaturing gradient gel electrophoresis (DGGE), did not reveal any variant in K-ras codons 12 and 13, which are the most frequently involved codons among the ras genes mutated in various types of human tumors. Analysis of the DNA extracted from four cell lines carrying known K-ras mutational alleles showed typically positive DGGE patterns. Thus, on the whole, the conclusions of this study and of previous studies using the same biological material are consistent with the occurrence of DNA adducts in human atherosclerotic lesions but in the absence of p53 involvement or of K-ras mutations in codons 12 and 13. The search for candidate genes which may possibly be involved in the atherogenetic process warrants further studies.
Insights
This study found no K-ras mutations in atherosclerotic lesions, suggesting these cancer biomarkers are not involved in atherosclerosis development. Further research is needed to identify atherogenesis-related genes.
Area of Science:
- Molecular Biology
- Cardiovascular Research
- Oncology
Background:
- Atherosclerosis is a complex cardiovascular disease.
- Cancer biomarkers are being investigated for roles in atherogenesis.
- K-ras mutations are common in human tumors.
Purpose of the Study:
- To investigate the presence of K-ras mutations in smooth muscle cells from human abdominal aorta atherosclerotic lesions.
- To determine if K-ras mutations, specifically in codons 12 and 13, are associated with atherosclerosis.
Main Methods:
- DNA extraction from 32 surgical specimens of atherosclerotic lesions.
- Polymerase Chain Reaction (PCR)-based denaturing gradient gel electrophoresis (DGGE) for molecular analysis.
- Analysis of known K-ras mutational alleles in cell lines as controls.
Main Results:
- No K-ras mutations were detected in codons 12 and 13 of DNA from atherosclerotic lesions.
- DGGE analysis confirmed the methodology's ability to detect K-ras mutations using control cell lines.
- Findings align with previous studies indicating DNA adducts but not p53 or K-ras mutations in lesions.
Conclusions:
- K-ras mutations in codons 12 and 13 do not appear to be involved in human atherogenesis.
- The study supports the presence of DNA adducts in atherosclerotic lesions.
- Further investigation into other candidate genes is warranted to understand atherogenesis.