Expressions of isopeptide bonds and corneodesmosin in middle ear cholesteatoma

Koizumi, Hiroki; Kawaguchi, Rintaro; Ohkubo, Jun‐ichi; Ikezaki, Shoji; Kitamura, Takuro; Hohchi, Nobusuke; Hashida, Koichi; Suzuki, Hideaki

doi:10.1111/coa.12703

Cited by 5 publications

(2 citation statements)

References 21 publications

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“…In the last decade it has become obvious that there is not one gene suitable for use of normalisation for all tissue types [40]. In the field of middle ear research many different genes have been used, such as B2M [52], PPIA [9], ACTB [33,34], HPRT1 [35] and GAPDH [36][37][38][39]. Interestingly, all articles published to this date use only one gene for normalisation, even though it has been abundantly shown that this is not sufficient for reliable analysis [40].…”

Section: Plos Onementioning

confidence: 99%

“…QPCR has been used in several studies related to diseases of the middle ear. However, there seems to be little consensus as to which reference genes to use for normalisation, leading to the use of several different genes without proper establishment of which genes are appropriate [9,[31][32][33][34][35][36][37][38][39]. To be able to obtain reliable results the use of stably expressed reference genes for normalisation is essential [40].…”

Section: Introductionmentioning

confidence: 99%

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

Drakskog¹,

Klerk²,

Westerberg³

et al. 2020

PLoS ONE

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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.

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