MiR-183 family regulates chloride intracellular channel 5 expression in inner ear hair cells

• Published in: Toxicology in vitro Vol. 27; no. 1; pp. 486 - 491
• Main Authors: Gu, Chunhui, Li, Xudong, Tan, Qiang, Wang, Zhi, Chen, Limei, Liu, Yimin
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.02.2013
• Subjects: Animals; Cell Line; Cell Line, Tumor; Chloride Channels - genetics; Chloride Channels - metabolism; CLIC5; Hair Cells, Auditory, Inner - metabolism; Humans; Inner ear hair cells; Mice; MicroRNAs - genetics; miR-183 family; CLIC5; miR-183 family; Inner ear hair cells
• ISSN: 0887-2333; 1879-3177
• DOI: 10.1016/j.tiv.2012.07.008

► MiR-96/-182 and CLIC5 are co-expressed in HEI-OC1 cells. ► CLIC5 expression was downregulated in HEI-OC1 after miR-96/-182 were transfected. ► CLIC5 is directly regulated by miR-96/-182. The roles of miRNAs in the onset of hearing and deafness are beginning to be revealed. Although there has been no reported link between chloride intracellular channel 5 (CLIC5) and the miR-183 family to date, we here present evidence that they are co-expressed in the inner ear and have functions that are related to stereocilia. Moreover, CLIC5 contains a single predicted and highly conserved miR-96/-182 binding site within its 3′-UTR. Our current results further show that miR-96/-182 and CLIC5 are co-expressed in HEI-OC1 cells, in which two isoforms of the CLIC5 protein exist. Furthermore, miR-96 and miR-182 were found to be specifically overexpressed in HEI-OC1 cells into which mimics of these molecules had been transfected by liposomes causing the downregulation of CLIC5 at both the mRNA and protein levels. Finally, miR-96/-182 specifically downregulate the expression of the luciferase reporter gene which was cloned into a mouse CLIC5 3′-UTR fragment containing the wild-type miR-96/-182 target sequence. Our findings thus suggest that CLIC5 is directly regulated by miR-96 and miR-182 and that the target sequence in this regard is located between nucleotides 760–766 within the CLIC5 3′-UTR.