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Cholesteatoma gene expression of matrix metalloproteinases and their inhibitors by RT-PCR
Carlos Eduardo Borges Rezende1, Ricardo Peres do Souto, Priscila Bogar Rapoport
1Medical Faculty of ABC. carlosbrezende@ig.com.br
Unlabelled:
Acquired middle ear cholesteatoma is a benign keratinizing hyperproliferative squamous epithelial lesion that may result in the destruction of the bone structures surrounding the temporal bone. Recent studies show that variations in cellular production of matrix metalloproteinases (MMPs) and their specific inhibitors (TIMPs) contribute to the pathophysiology of cholesteatoma.
Objective:
This study aims to analyze the use of RNA amplification tests to evaluate the expression of MMP and TIMP isoforms in cholesteatomas and their correlation with disease severity.
Materials And Methods:
This is a prospective study. Nineteen cholesteatoma cases at different stages were selected. RNA collected from biopsy specimens was submitted to reverse transcription polymerase chain reaction (RT-PCR) for semiquantitative amplification of MMP2, MMP3, MMP9, MMP13 and TIMP1.
Results:
Six cholesteatomas were positive for at least one of the studied genes. Four samples amplified a single gene (MMP2 or MMP13) and two samples amplified three genes (MMP2, TIMP1 and MMP3 or MMP13). No sample amplified MMP9.
Conclusion:
RT-PCR can be used to assess MMP and TIMP gene expression in cholesteatomas despite technical difficulties. Gene expression profiles could not be related to disease severity.
Insights
RNA amplification tests can detect matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) in cholesteatomas. However, gene expression profiles did not correlate with disease severity in this study.
Area of Science:
- Otolaryngology
- Molecular Biology
- Pathophysiology
Background:
- Acquired middle ear cholesteatoma is a benign lesion that can destroy surrounding bone.
- Matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) are implicated in cholesteatoma development.
Purpose of the Study:
- To evaluate RNA amplification tests for assessing MMP and TIMP gene expression in cholesteatomas.
- To correlate gene expression profiles with cholesteatoma disease severity.
Main Methods:
- Prospective study of 19 cholesteatoma cases.
- RNA extraction from biopsy specimens.
- Reverse transcription polymerase chain reaction (RT-PCR) for semiquantitative amplification of MMP2, MMP3, MMP9, MMP13, and TIMP1.
Main Results:
- Six cholesteatomas showed positive amplification for at least one gene.
- Four samples amplified a single gene (MMP2 or MMP13).
- Two samples amplified multiple genes (MMP2, TIMP1, MMP3 or MMP13).
- MMP9 was not detected in any sample.
Conclusions:
- RT-PCR is a viable method for assessing MMP and TIMP gene expression in cholesteatomas, despite technical challenges.
- No correlation was found between MMP and TIMP gene expression profiles and cholesteatoma disease severity.
