Objectives
The goal of this study was to identify novel regulatory mechanisms controlling the growth and proliferation of cholesteatoma. Specifically, the potential role of microRNAs, regulators of protein translation, was studied in cholesteatoma.
Study Design
This study represents a molecular biological investigation characterizing and comparing microRNA and protein expression in cholesteatoma and normal post-auricular skin.
Methods
Cholesteatoma and normal skin were taken from patients at the time of surgery. Tissue was processed for RNA and protein extraction. Real-time RT-PCR was used to assess levels of human microRNAs, RT-PCR was used to confirm the presence of upstream regulators, and western blot analyses were used to assess levels of downstream target proteins.
Results
Among the microRNAs investigated, human microRNA-21 (hsa-mir-21) showed a 4.4-fold higher expression in cholesteatoma as compared to normal skin (p=0.0011). The downstream targets of hsa-mir-21, PTEN and PDCD4, were found to be greatly reduced in 3 of 4 cholesteatoma samples. Proposed upstream regulators of hsa-mir-21 expression (CD14, IL-6R, gp130 and STAT3) were present in all cholesteatoma tissues.
Conclusions
MicroRNAs represent powerful regulators of protein translation and their dysregulation has been implicated in many neoplastic diseases. This study specifically identified up-regulation of hsa-mir-21 concurrent with down-regulation of potent tumor suppressor proteins, PTEN and PDCD4. These proteins control aspects of apoptosis, proliferation, invasion and migration. The results of this study were used to develop a model for cholesteatoma proliferation through microRNA dysregulation. This model can serve as a template for further study into potential RNA-based therapies for the treatment of cholesteatoma.