Autophagy Is Deficient and May be Negatively Regulated by SERPINB3 in Middle Ear Cholesteatoma. Issue 7 (August 2020)

• Record Type: Journal Article
• Authors: Ho, Kuen-Yao; Huang, Chih-Jen; Hung, Chih-Chang; Wu, Yu-Rong; Chiu, Chien-Chih; Chien, Chen-Yu; Wang, Hsun-Mo; Chang, Ning-Chia; Lin, I-Ling; Chen, Jeff Yi-Fu
• Is Part Of: Otology & neurotology. Volume 41:Issue 7(2020)
• Publication Date: 2020-08
• Subjects: Autophagy -- Cholesteatoma -- 1A/1B-light chain 3 -- Serine proteinase inhibitor, clade B, member 3; Otology -- Periodicals; Ear -- Diseases -- Periodicals; Skull base -- Surgery -- Periodicals; 617.8005
• DOI: 10.1097/MAO.0000000000002690
• Languages: English
• ISSNs: 1531-7129

Abstract

Abstract : Hypothesis: Whereas autophagy has been linked to various human diseases, whether it also plays a role in cholesteatoma is virtually unknown. This study aimed to investigate the activity and regulation of autophagy in cholesteatoma. Background: The treatment of middle ear cholesteatoma has been challenging due to an insufficient understanding of the underlying disease mechanism. Methods: Expression of microtubule-associated protein 1A/1B-light chain 3 (LC3), the autophagy protein marker, and phosphorylated Akt (p-Akt), and mammalian target of rapamycin (p- mTOR ), the known autophagy regulators, in fresh retroauricular skin and cholesteatoma tissue samples was analyzed by immunoblotting. The results were further confirmed by immunohistochemistry and statistical analyses. Cell proliferation of primary retroauricular skin- and cholesteatoma-derived fibroblasts was evaluated by methyl thiazol tetrazolium (MTT) assay. Ectopic expression of serine proteinase inhibitor, clade B, member 3 (SERPINB3) in the fibroblasts was achieved by electroporation and the expression was detected by immunoblotting. Results: LC3 expression was significantly decreased in cholesteatoma in most of the 15 paired retroauricular skin/cholesteatoma tissue samples. However, p-Akt and p-mTOR expression in the cholesteatoma samples was not significantly different from that in the control subjects. Immunohistochemical studies further demonstrated an inverse correlation between LC3 expression and cholesteatoma. The cholesteatoma fibroblasts proliferated faster than the retroauricular skin fibroblasts, and had higher SERPINB3 but lower LC3 expression. Furthermore, overexpression of SERPINB3 in the retroauricular skin fibroblasts enhanced cell proliferation and downregulated LC3 expression. Conclusion: Autophagy is significantly suppressed in cholesteatoma tissues, which may not involve the Akt/mTOR signaling pathway. More importantly, SERPINB3 may promote cell proliferation and negatively regulate autophagy in cholesteatoma fibroblasts. Together, these findings warrant further investigation into the pathogenic mechanism of cholesteatoma.