Expression of female sex hormone receptors in human middle-ear cholesteatomas.

The purpose of this study was to establish the eventual presence of progesterone receptor (PGR) and oestrogen receptor (EGR) in human middle-ear cholesteatoma (MECh) tissues and to compare their expression between male and female patients. An immunohistochemical technique was employed for detection of PGR- and EGR-specific immunoreactivity in MECh samples using formalin-fixed paraffin-embedded tissue sections. The positive results were verified with reverse transcriptase polymerase chain reaction (RT-PCR). The morphological study revealed stable expression of PGR in suprabasal layers of all cholesteatoma samples. Weaker immunoreactivity for PGR was demonstrated in external auditory canal skin (EACS) samples in comparison with MECh, while PGR-specific staining was not observed in retroauricular skin (RAS) samples. EGR was detected only at mRNA levels. Stronger expression of EGR PCR products was disclosed in female cholesteatoma samples, while PGR mRNA was predominantly detected in male cholesteatoma specimens. Our preliminary experimental results give us ground to assume that female sex hormones may stimulate proliferation and affect differentiation of MECh keratinocytes

Nucleoplasm staining patterns and cell cycle-associated expression of Ki-67 in middle ear cholesteatoma.

OBJECTIVES: To compare Ki-67 expression patterns in middle ear cholesteatoma with the corresponding retroauricular and external auditory canal skins, and to determine the cell cycle-dependent localization of Ki-67. MATERIAL AND METHODS: MIB-1 monoclonal antibody was used for comparative assessment of proliferative activity of middle ear cholesteatoma, external auditory canal skin, and retroauricular skin samples on formalin-fixed paraffin-embedded tissue sections. Primary keratinocytes from cholesteatoma tissue were isolated and subjected to kinetic analysis of the cell cycle. RESULTS: Higher proliferative activity was established in cholesteatoma in comparison with retroauricular and external auditory canal skins. Three different staining patterns have been described. Kinetic analysis revealed continuous expression of Ki-67 during all active phases of the cell cycle and remained "silent" in resting cells. CONCLUSION: The established correlation between the staining patterns and cell cycle-associated expression of Ki-67 specifies Ki-67 as a reliable and stable marker of proliferation for middle ear cholesteatoma

Expression of the gap junction proteins connexin 26 and connexin 43 in human middle ear cholesteatoma.

CONCLUSION: The results of this study showed upregulated expression and a change in localization of both connexin 43 (Cx43) and Cx26 in human middle ear cholesteatoma compared to those in normal retroauricular skins (RASs) and ear canal skins (ECSs). This suggests that perturbations of intercellular communication through gap junctions may be associated with the pathology of human cholesteatomas. OBJECTIVE: Cholesteatomas in the middle ear require intercellular signal exchange through gap junctions as well as intracellular signal pathways for the hyperproliferation and differentiation of epithelial cells. Cx is a gap junction protein involved in intercellular communication. The objective of this study was to analyze the expression and possible roles of Cx43 and Cx26 in human cholesteatoma compared to normal epithelium. MATERIAL AND METHODS: Ten RASs, 10 ECSs and 10 cholesteatomas were obtained during middle ear operations. Immunohistochemical staining, Western blotting and reverse transcriptase polymerase chain reaction (RT-PCR) were used to detect Cx43 and Cx26. The expression patterns of Cx43 and Cx26 were also compared with that of the proliferation marker Ki67. RESULTS: In human cholesteatomas, Cx43 was expressed in whole suprabasal layers, except in the basal layer, and Cx26 was usually expressed in the suprabasal and basal layers. However, normal RASs showed weak expression of Cx43 in the upper spinosal and granular layers (with no expression in the basal layers) and restricted localization of Cx26 in the basal layer. The expression of Cx43 and Cx26 in ECSs was weak but showed similar patterns to that of cholesteatoma. RT-PCR and Western blotting showed that the expression of Cx43 and Cx26 was higher in cholesteatoma than in RASs. Epithelial cells expressing Cx43 and Cx26 in cholesteatoma were not exactly identical to Ki67-expressing cells on immunohistochemical staining