Using CCM and DHPLC to detect mutations in the glucocorticoid receptor in atherosclerosis: a comparison
1Mutation Research Centre, St. Vincent's Hospital, 41 Victoria Parade, Victoria 3065, Fitzroy, Australia.
Abstract:
Growing evidence suggests that restenosis may be caused by a failure in growth inhibitory and apoptotic systems that would normally mediate lesion regression. One such inhibitory system is the glucocorticoid receptor. This paper develops, assesses and compares chemical cleavage of mismatch (CCM) and denaturing high-performance liquid chromatography (DHPLC) for their utility in detecting mutations in this system. The results of the two methods correlated in 74% of cases in a cohort of endarterectomy patients studied by these two methods.
Insights
Glucocorticoid receptor mutations may cause restenosis due to failed lesion regression. Chemical cleavage of mismatch (CCM) and denaturing high-performance liquid chromatography (DHPLC) methods showed 74% correlation in detecting these mutations in endarterectomy patients.
Area of Science:
- Molecular biology
- Genetics
- Cardiovascular research
Background:
- Restenosis, a cardiovascular condition, may stem from impaired growth inhibition and apoptosis, hindering lesion regression.
- The glucocorticoid receptor system plays a crucial role in mediating these inhibitory processes.
- Identifying mutations within the glucocorticoid receptor is vital for understanding restenosis pathogenesis.
Purpose of the Study:
- To develop and assess chemical cleavage of mismatch (CCM) and denaturing high-performance liquid chromatography (DHPLC) for detecting glucocorticoid receptor mutations.
- To compare the utility and correlation of CCM and DHPLC in identifying these mutations.
- To investigate the role of glucocorticoid receptor mutations in endarterectomy patients.
Main Methods:
- Chemical cleavage of mismatch (CCM) assay development and application.
- Denaturing high-performance liquid chromatography (DHPLC) analysis for mutation detection.
- Comparative analysis of CCM and DHPLC results in a patient cohort.
Main Results:
- Both CCM and DHPLC were assessed for their effectiveness in mutation detection.
- A significant correlation was observed between the results obtained from CCM and DHPLC.
- The two methods showed a 74% correlation in identifying mutations in the studied endarterectomy patient cohort.
Conclusions:
- CCM and DHPLC are viable methods for detecting mutations in the glucocorticoid receptor system.
- The findings support the potential role of glucocorticoid receptor dysfunction in restenosis.
- Further research is warranted to elucidate the precise mechanisms linking these mutations to cardiovascular lesion progression.
