title: Mir-27a promotes apoptosis of cochlear sensory epithelium in Cx26 knockout mice.
abstract: To investigate the underlying molecular mechanism for connexin 26 (Cx26) knockout-induced apoptosis, we performed TUNEL assays to detect apoptosis in the cochlear sensory epithelium in Cx26 knockout mice. We also compared the miRNA expression profiles of Cx26 knockout and wild-type mice using microarray technology and bioinformatic analyses. Real-time PCR, luciferase reporter gene assays, and scala media microinjections were performed to identify the effect of a specific miRNA and its targets. The results showed that apoptosis increased in the cochlear sensory epithelium of Cx26 knockout mice. The abnormal expression of mir-27a and sgk1 in Cx26 knockout mice was verified with real-time PCR. Luciferase reporter gene assays showed that overexpression of mir-27a significantly decreased sgk1 reporter gene activity; an inhibitor of mir-27a blocked the effect. Mir-27a lentivirus also inhibited sgk1 expression in cultured cochlear tissue. Mir-27a shRNA treatment inhibited Cx26 knockout-induced apoptosis in the cochlear sensory epithelium of mice and increased the expression of sgk1 mRNA. Thus, mir-27a was identified as an apoptotic molecule that participates in Cx26 knockout-induced apoptosis in the cochlear sensory epithelium of mice by downregulating sgk1 expression.
authors.forename: Yunfeng
authors.initials: Y
authors.lastname: Wang
authors.forename: Chen
authors.initials: C
authors.lastname: Lin
authors.forename: Yingzi
authors.initials: Y
authors.lastname: He
authors.forename: Ao
authors.initials: A
authors.lastname: Li
authors.forename: Wenli
authors.initials: W
authors.lastname: Ni
authors.forename: Shan
authors.initials: S
authors.lastname: Sun
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Gu
authors.forename: Jian
authors.initials: J
authors.lastname: Li
authors.affiliation: Department of Otolaryngology, Affiliated Eye and ENT Hospital of Fudan University, Building 9 Room 611, 83 Fenyang Road, Xuhui District, Shanghai 200031, China, huawli@sina.cn.
authors.forename: Huawei
authors.initials: H
authors.lastname: Li
mesh_terms: D020413:3' Untranslated Regions; D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D003051:Cochlea / Q000166:cytology*; D000072259:Connexin 26; D017630:Connexins / Q000235:genetics*; D004847:Epithelial Cells / Q000166:cytology; D057809:HEK293 Cells; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics; D051379:Mice; D018345:Mice, Knockout; D035683:MicroRNAs / Q000502:physiology*; D017346:Protein Serine-Threonine Kinases / Q000235:genetics; D000099184:Serum-Glucocorticoid Regulated Kinases
chemical_list: D020413:3' Untranslated Regions; D000072259:Connexin 26; D017630:Connexins; D017874:Immediate-Early Proteins; D035683:MicroRNAs; D017346:Protein Serine-Threonine Kinases; D000099184:Serum-Glucocorticoid Regulated Kinases; C000607042:GJB2 protein, human; C000607043:Gjb2 protein, mouse; C540674:Mirn27 microRNA, mouse