Extensive qPCR analysis reveals altered gene expression in middle ear mucosa from cholesteatoma patients

Drakskog, CeciliaThe Division of ENT Diseases, CLINTEC, Karolinska Institutet, Stockholm, Sweden (author)

Funding agencies: Swedish research councilSwedish Research Council
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.

de Klerk, NeleThe Division of ENT Diseases, CLINTEC, Karolinska Institutet, Stockholm, Sweden (author)
Westerberg, Johanna,1971-Linköpings universitet,Avdelningen för sinnesorgan och kommunikation,Medicinska fakulteten,Region Östergötland, Öron- näsa- och halskliniken,Division of Neuro and Inflammation Science,Department of Clinical and Experimental Medicine, Linköping University, Linköping, Sweden; Department of Otorhinolaryngologyin Linköping, Anaesthetics, Operations and Specialty Surgery Centre, Region Östergötland, Linköping, Sweden(Swepub:liu)johwe74 (author)
Mäki-Torkko, Elina,Professor,1961-Örebro universitet,Linköpings universitet,Avdelningen för sinnesorgan och kommunikation,Medicinska fakulteten,Region Östergötland, Öron- näsa- och halskliniken,Faculty of Medicine and Health, Örebro University,Institutionen för medicinska vetenskaper,Division of Neuro and Inflammation Science,Department of Clinical and Experimental Medicine, Linköping University, Linköping, Sweden; Department of Otorhinolaryngologyin Linköping, Anaesthetics, Operations and Specialty Surgery Centre, Region Östergötland, Linköping, Sweden(Swepub:oru)emo (author)
Georén, Susanna KumlienKarolinska Institutet (author)
Cardell, Lars OlafKarolinska Institutet (author)
The Division of ENT Diseases, CLINTEC, Karolinska Institutet, Stockholm, SwedenAvdelningen för sinnesorgan och kommunikation (creator_code:org_t)

By the author/editor: Drakskog, Cecili ...; de Klerk, Nele; Westerberg, Joha ...; Mäki-Torkko, Eli ...; Georén, Susanna ...; Cardell, Lars Ol ...

About the subject

MEDICAL AND HEALTH SCIENCES: MEDICAL AND HEAL ...; and Basic Medicine; and Medical Genetics

MEDICAL AND HEALTH SCIENCES: MEDICAL AND HEAL ...; and Clinical Medicin ...; and Otorhinolaryngol ...

By the university: Linköping University; Örebro University; Karolinska Institutet

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