Extensive qPCR analysis reveals altered gene expression in middle ear mucosa from cholesteatoma patients.
Cecilia DrakskogNele de KlerkJohanna WesterbergElina Mäki-TorkkoSusanna Kumlien GeorénLars-Olaf CardellPublished in: PloS one (2020)
The middle ear is a small and hard to reach compartment, limiting the amount of tissue that can be extracted and the possibilities for studying the molecular mechanisms behind diseases like cholesteatoma. In this paper 14 reference gene candidates were evaluated in the middle ear mucosa of cholesteatoma patients and two different control tissues. ACTB and GAPDH were shown to be the optimal genes for the normalisation of target gene expression when investigating middle ear mucosa in multiplex qPCR analysis. Validation of reference genes using c-MYC expression confirmed the suitability of ACTB and GAPDH as reference genes and showed an upregulation of c-MYC in middle ear mucosa during cholesteatoma. The occurrence of participants of the innate immunity, TLR2 and TLR4, were analysed in order to compare healthy middle ear mucosa to cholesteatoma. Analysis of TLR2 and TLR4 showed variable results depending on control tissue used, highlighting the importance of selecting relevant control tissue when investigating causes for disease. It is our belief that a consensus regarding reference genes and control tissue will contribute to the comparability and reproducibility of studies within the field.
Keyphrases
- gene expression
- genome wide
- end stage renal disease
- inflammatory response
- ejection fraction
- immune response
- dna methylation
- genome wide identification
- newly diagnosed
- poor prognosis
- prognostic factors
- cell proliferation
- signaling pathway
- peritoneal dialysis
- risk assessment
- nuclear factor
- clinical practice
- patient reported
- binding protein