miR‑125/CDK2 axis in cochlear progenitor cell proliferation

Peng,Tao; Peng,Jing-Jing; Miao,Gang-Yong; Tan,Zhi-Qiang; Liu,Bin; Zhou,En

doi:10.3892/mmr.2020.11741

miR‑125/CDK2 axis in cochlear progenitor cell proliferation

Authors:
- Tao Peng
- Jing-Jing Peng
- Gang-Yong Miao
- Zhi-Qiang Tan
- Bin Liu
- En Zhou
View Affiliations

Affiliations: Department of Otolaryngology and Head and Neck Surgery, Hunan Provincial People's Hospital, First Affiliated Hospital of Hunan Normal University, Changsha, Hunan 410007, P.R. China, Department of Obstetrics and Gynecology, Changsha Maternal and Child Health Care Hospital, Changsha, Hunan 410005, P.R. China
Published online on: December 1, 2020 https://doi.org/10.3892/mmr.2020.11741
Article Number: 102
Copyright: © Peng et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Hearing loss ranks fourth among the principal causes of disability worldwide, and manipulation of progenitor cells may be a key strategy for hair cell regeneration. The present study investigated the role and mechanism of miR‑125 on the proliferation of cochlear progenitor cells (CPCs). CPCs were isolated from the cochleae of neonatal rats, and their morphology was observed. Furthermore, the differentiation ability of CPCs was determined by assessing the expression of 5‑bromodeoxyuridine (BrdU), nestin and myosin VII by immunofluorescence. The expression levels of miR‑125 and cyclin‑dependent kinase 2 (CDK2) as well as the cell proliferation of CPCs were assessed. In addition, following gain‑ and loss‑of‑function assays, the cell cycle was examined by flow cytometry, and the expression levels of miR‑125, CDK2, proliferating cell nuclear antigen (PCNA) and nestin were determined by reverse transcription‑quantitative PCR and western blotting. The binding sites between miR‑125 and CDK2 were predicted by TargetScan and identified by the dual luciferase reporter assay. The results demonstrated that different types of progenitor spheres were observed from CPCs with positive expression of BrdU, nestin and myosin VII. Following in vitro incubation for 2, 4 and 7 days, the spheres were enlarged, and CPC proliferation gradually increased and reached a plateau after further incubation for 3 days. Furthermore, the expression levels of nestin and PCNA in CPCs increased and then decreased during in vitro incubation for 2, 4 and 7 days. Following this incubation, the expression levels of miR‑125 in CPCs decreased; thereafter, its expression increased, and the expression pattern was different from that of CDK2. In addition, miR‑125 overexpression in CPCs decreased the expression of CDK2 and the number of cells in the S phase. Different expression patterns were found in CPCs in response to the miR‑125 knockdown. In addition, miR‑125 directly targeted CDK2. Simultaneous knockdown of miR‑125 and CDK2 enhanced CPC proliferation compared with CDK2 knockdown alone. Taken together, the findings from the present study suggested that miR‑125 may inhibit CPC proliferation by downregulating CDK2. The present study may provide a novel therapeutic direction for treatment of hearing loss.

View Figures

View References

1	Cunningham LL and Tucci DL: Hearing loss in adults. N Engl J Med. 377:2465–2473. 2017. View Article : Google Scholar
2	Kim YR, Baek JI, Kim SH, Kim MA, Lee B, Ryu N, Kim KH, Choi DG, Kim HM, Murphy MP, et al: Therapeutic potential of the mitochondria-targeted antioxidant MitoQ in mitochondrial-ROS induced sensorineural hearing loss caused by Idh2 deficiency. Redox Biol. 20:544–555. 2019. View Article : Google Scholar
3	Tang M, Yan X, Tang Q, Guo R, Da P and Li D: Potential application of electrical stimulation in stem cell-based treatment against hearing loss. Neural Plast. 2018:95063872018. View Article : Google Scholar
4	Lee MY and Park YH: Potential of gene and cell therapy for inner ear hair cells. Biomed Res Int. 2018:81376142018. View Article : Google Scholar
5	Roccio M, Perny M, Ealy M, Widmer HR, Heller S and Senn P: Molecular characterization and prospective isolation of human fetal cochlear hair cell progenitors. Nat Commun. 9:40272018. View Article : Google Scholar
6	Lin SCY, Thorne PR, Housley GD and Vlajkovic SM: Purinergic signaling and aminoglycoside ototoxicity: The opposing roles of P1 (Adenosine) and P2 (ATP) receptors on cochlear hair cell survival. Front Cell Neurosci. 13:2072019. View Article : Google Scholar
7	Revuelta M, Santaolalla F, Arteaga O, Alvarez A, Sanchez-Del-Rey A and Hilario E: Recent advances in cochlear hair cell regeneration-A promising opportunity for the treatment of age-related hearing loss. Ageing Res Rev. 36:149–155. 2017. View Article : Google Scholar
8	Youm I and Li W: Cochlear hair cell regeneration: An emerging opportunity to cure noise-induced sensorineural hearing loss. Drug Discov Today. 23:1564–1569. 2018. View Article : Google Scholar
9	Huh SH, Warchol ME and Ornitz DM: Cochlear progenitor number is controlled through mesenchymal FGF receptor signaling. Elife. 4:e059212015. View Article : Google Scholar
10	Mittal R, Liu G, Polineni SP, Bencie N, Yan D and Liu XZ: Role of microRNAs in inner ear development and hearing loss. Gene. 686:49–55. 2019. View Article : Google Scholar
11	Pang J, Xiong H, Yang H, Ou Y, Xu Y, Huang Q, Lai L, Chen S, Zhang Z, Cai Y and Zheng Y: Circulating miR-34a levels correlate with age-related hearing loss in mice and humans. Exp Gerontol. 76:58–67. 2016. View Article : Google Scholar
12	La Torre A, Georgi S and Reh TA: Conserved microRNA pathway regulates developmental timing of retinal neurogenesis. Proc Natl Acad Sci USA. 110:E2362–E2370. 2013. View Article : Google Scholar
13	Li L, Wang Q, Yuan Z, Chen A, Liu Z, Wang Z and Li H: LncRNA-MALAT1 promotes CPC proliferation and migration in hypoxia by up-regulation of JMJD6 via sponging miR-125. Biochem Biophys Res Commun. 499:711–718. 2018. View Article : Google Scholar
14	Spencer SL, Cappell SD, Tsai FC, Overton KW, Wang CL and Meyer T: The proliferation-quiescence decision is controlled by a bifurcation in CDK2 activity at mitotic exit. Cell. 155:369–383. 2013. View Article : Google Scholar
15	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar
16	Song YL, Tian KY, Mi WJ, Ding ZJ, Qiu Y, Chen FQ, Zha DJ and Qiu JH: Decreased expression of TERT correlated with postnatal cochlear development and proliferation reduction of cochlear progenitor cells. Mol Med Rep. 17:6077–6083. 2018.
17	Zhang Y, Guo L, Lu X, Cheng C, Sun S, Li W, Zhao L, Lai C, Zhang S, Yu C, et al: Characterization of Lgr6⁺ cells as an enriched population of hair cell progenitors compared to Lgr5⁺ cells for hair cell generation in the neonatal mouse cochlea. Front Mol Neurosci. 11:1472018. View Article : Google Scholar
18	Gao J, Wan F, Tian M, Li Y, Li Y, Li Q, Zhang J, Wang Y, Huang X, Zhang L and Si Y: Effects of ginsenoside-Rg1 on the proliferation and gliallike directed differentiation of embryonic rat cortical neural stem cells in vitro. Mol Med Rep. 16:8875–8881. 2017. View Article : Google Scholar
19	Takeda H, Dondzillo A, Randall JA and Gubbels SP: Challenges in cell-based therapies for the treatment of hearing loss. Trends Neurosci. 41:823–837. 2018. View Article : Google Scholar
20	Hei R, Chen J, Qiao L, Li X, Mao X, Qiu J and Qu J: Dynamic changes in microRNA expression during differentiation of rat cochlear progenitor cells in vitro. Int J Pediatr Otorhinolaryngol. 75:1010–1014. 2011. View Article : Google Scholar
21	Huyghe A, Van den Ackerveken P, Sacheli R, Prevot PP, Thelen N, Renauld J, Thiry M, Delacroix L, Nguyen L and Malgrange B: MicroRNA-124 regulates cell specification in the cochlea through modulation of Sfrp4/5. Cell Rep. 13:31–42. 2015. View Article : Google Scholar
22	Wang XR, Zhang XM, Du J and Jiang H: MicroRNA-182 regulates otocyst-derived cell differentiation and targets T-box1 gene. Hear Res. 286:55–63. 2012. View Article : Google Scholar
23	Yang M, Tang X, Wang Z, Wu X, Tang D and Wang D: miR-125 inhibits colorectal cancer proliferation and invasion by targeting TAZ. Biosci Rep. 39:BSR201901932019. View Article : Google Scholar
24	Li L, Zhang M, Chen W, Wang R, Ye Z, Wang Y, Li X and Cai C: LncRNA-HOTAIR inhibition aggravates oxidative stress-induced H9c2 cells injury through suppression of MMP2 by miR-125. Acta Biochim Biophys Sin (Shanghai). 50:996–1006. 2018. View Article : Google Scholar
25	Tu XM, Gu YL and Ren GQ: miR-125a-3p targetedly regulates GIT1 expression to inhibit osteoblastic proliferation and differentiation. Exp Ther Med. 12:4099–4106. 2016. View Article : Google Scholar
26	Song Z, Jadali A, Fritzsch B and Kwan KY: NEUROG1 regulates CDK2 to promote proliferation in Otic progenitors. Stem Cell Reports. 9:1516–1529. 2017. View Article : Google Scholar
27	Kaur S, Abu-Shahba AG, Paananen RO, Hongisto H, Hiidenmaa H, Skottman H, Seppanen-Kaijansinkko R and Mannerström B: Small non-coding RNA landscape of extracellular vesicles from human stem cells. Sci Rep. 8:155032018. View Article : Google Scholar
28	Malgrange B, Knockaert M, Belachew S, Nguyen L, Moonen G, Meijer L and Lefebvre PP: The inhibition of Cyclin-dependent kinases induces differentiation of supernumerary hair cells and Deiters' cells in the developing organ of Corti. FASEB J. 17:2136–2138. 2003. View Article : Google Scholar
29	Shenoy A, Danial M and Blelloch RH: Let-7 and miR-125 cooperate to prime progenitors for astrogliogenesis. EMBO J. 34:1180–1194. 2015. View Article : Google Scholar