miR‑200c‑3p regulates the proliferation and apoptosis of human trabecular meshwork cells by targeting PTEN

Shen,Yifei; Zhu,Yin; Rong,Fei

doi:10.3892/mmr.2020.11198

miR‑200c‑3p regulates the proliferation and apoptosis of human trabecular meshwork cells by targeting PTEN

Authors:
- Yifei Shen
- Yin Zhu
- Fei Rong
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Affiliations: Department of Ophthalmology, Wujiang First People's Hospital, Suzhou, Jiangsu 215200, P.R. China
Published online on: June 1, 2020 https://doi.org/10.3892/mmr.2020.11198
Pages: 1605-1612

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Abstract

Glaucoma is an optic neuropathy that may lead to visual field loss and blindness. Human trabecular meshwork (HTM) cell dysfunction increases intraocular pressure, which leads to glaucoma. microRNAs (miRs) are single‑stranded non‑coding RNAs that regulate cellular processes in HTM cells. The aim of the present study was to evaluate the role of miR‑200c‑3p in HTM cells. HTM cells were treated with 300 µM H2O2, and the expression of miR‑200c‑3p was measured by reverse transcription‑quantitative polymerase chain reaction. The expression of miR‑200c‑3p was significantly downregulated under oxidative stress. Phosphatase and tensin homolog (PTEN) was predicted to be a potential target of miR‑200c‑3p using TargetScan, and this was confirmed by a dual‑luciferase reporter assay. The expression of PTEN was upregulated in H2O2‑treated HTM cells. In addition, PTEN expression was negatively regulated by miR‑200c‑3p. Then, cell proliferation and apoptosis were measured by Cell Counting Kit‑8 and flow cytometry assays, respectively. The results demonstrated that the overexpression of miR‑200c‑3p enhanced cell proliferation and inhibited cell apoptosis, while PTEN reversed the effects of miR‑200c‑3p. In addition, miR‑200c‑3p suppressed the expression of PTEN, cleaved caspase‑3 and Bax, and improved the expression of phosphorylated (p)‑AKT, AKT and p‑serine/threonine‑protein kinase mTOR (mTOR), while PTEN attenuated these effects. Taken together, these data suggested that overexpression of miR‑200c‑3p targets PTEN to suppress cleaved caspase‑3 and Bax expression and activate the PTEN/AKT/mTOR signaling pathway, thereby resulting in the promotion of cell proliferation and the inhibition of cell apoptosis. These results revealed the role of miR‑200c‑3p in HTM cells and its underlying molecular mechanism, which may provide a potential target for the treatment of patients with glaucoma.

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1	Mantravadi AV and Vadhar N: Glaucoma. Prim Care. 42:437–449. 2015. View Article : Google Scholar : PubMed/NCBI
2	Nickells RW, Howell GR, Soto I and John SW: Under pressure: Cellular and molecular responses during glaucoma, a common neurodegeneration with axonopathy. Annu Rev Neurosci. 35:153–179. 2012. View Article : Google Scholar : PubMed/NCBI
3	Stamer WD and Clark AF: The many faces of the trabecular meshwork cell. Exp Eye Res. 158:112–123. 2017. View Article : Google Scholar : PubMed/NCBI
4	Porter K, Hirt J, Stamer WD and Liton PB: Autophagic dysregulation in glaucomatous trabecular meshwork cells. Biochim Biophys Acta. 1852:379–385. 2015. View Article : Google Scholar : PubMed/NCBI
5	Ruibin W, Zheng X, Chen J, Zhang X, Yang X and Lin Y: Micro RNA-1298 opposes the effects of chronic oxidative stress on human trabecular meshwork cells via targeting on EIF4E3. Biomed Pharmacother. 100:349–357. 2018. View Article : Google Scholar : PubMed/NCBI
6	Johnstone MA: The aqueous outflow system as a mechanical pump: Evidence from examination of tissue and aqueous movement in human and non-human primates. J Glaucoma. 13:421–438. 2004. View Article : Google Scholar : PubMed/NCBI
7	Bartel DP: MicroRNAs: Genomics, biogenesis, mechanism, and function. Cell. 116:281–297. 2004. View Article : Google Scholar : PubMed/NCBI
8	Pillai RS: MicroRNA function: Multiple mechanisms for a tiny RNA? RNA. 11:1753–1761. 2005. View Article : Google Scholar : PubMed/NCBI
9	Guo R, Shen W, Su C, Jiang S and Wang J: Relationship between the pathogenesis of glaucoma and miRNA. Ophthalmic Res. 57:194–199. 2017. View Article : Google Scholar : PubMed/NCBI
10	Gonzalez P, Li G, Qiu J, Wu J and Luna C: Role of microRNAs in the trabecular meshwork. J Ocul Pharmacol Ther. 30:128–137. 2014. View Article : Google Scholar : PubMed/NCBI
11	Feng X, Wang Z, Fillmore R and Xi Y: MiR-200, a new star miRNA in human cancer. Cancer Lett. 344:166–173. 2014. View Article : Google Scholar : PubMed/NCBI
12	Romano GL, Platania CB, Forte S, Salomone S, Drago F and Bucolo C: MicroRNA target prediction in glaucoma. Prog Brain Res. 220:217–240. 2015. View Article : Google Scholar : PubMed/NCBI
13	Luna C, Li G, Huang J, Qiu J, Wu J, Yuan F, Epstein DL and Gonzalez P: Regulation of trabecular meshwork cell contraction and intraocular pressure by miR-200c. PLoS One. 7:e516882012. View Article : Google Scholar : PubMed/NCBI
14	Kimura A, Namekata K, Guo X, Noro T, Harada C and Harada T: Targeting oxidative stress for treatment of glaucoma and optic neuritis. Oxid Med Cell Longev. 2017:28172522017. View Article : Google Scholar : PubMed/NCBI
15	Zhao J, Wang S, Zhong W, Yang B, Sun L and Zheng Y: Oxidative stress in the trabecular meshwork (Review). Int J Mol Med. 38:995–1002. 2016. View Article : Google Scholar : PubMed/NCBI
16	Saccà SC, Gandolfi S, Bagnis A, Manni G, Damonte G, Traverso CE and Izzotti A: The outflow pathway: A tissue with morphological and functional unity. J Cell Physiol. 231:1876–1893. 2016. View Article : Google Scholar : PubMed/NCBI
17	Agarwal V, Bell GW, Nam JW and Bartel DP: Predicting effective microRNA target sites in mammalian mRNAs. Elife. 4:2015. View Article : Google Scholar
18	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta DeltaC(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
19	Carlomosti F, D'Agostino M, Beji S, Torcinaro A, Rizzi R, Zaccagnini G, Maimone B, Di Stefano V, De Santa F, Cordisco S, et al: Oxidative stress-induced miR-200c disrupts the regulatory loop among SIRT1, FOXO1, and eNOS. Antioxid Redox Signal. 27:328–344. 2017. View Article : Google Scholar : PubMed/NCBI
20	Wang Y, Zhou H, Liu X, Han Y, Pan S and Wang Y: MiR-181a inhibits human trabecular meshwork cell apoptosis induced by H2O2 through the suppression of NF-κB and JNK pathways. Adv Clin Exp Med. 27:577–582. 2018. View Article : Google Scholar : PubMed/NCBI
21	Mutlu M, Raza U, Saatci Ö, Eyüpoğlu E, Yurdusev E and Şahin Ö: miR-200c: A versatile watchdog in cancer progression, EMT, and drug resistance. J Mol Med (Berl). 94:629–644. 2016. View Article : Google Scholar : PubMed/NCBI
22	Li G, Luna C, Qiu J, Epstein DL and Gonzalez P: Alterations in microRNA expression in stress-induced cellular senescence. Mech Ageing Dev. 130:731–741. 2009. View Article : Google Scholar : PubMed/NCBI
23	Milella M, Falcone I, Conciatori F, Cesta Incani U, Del Curatolo A, Inzerilli N, Nuzzo CM, Vaccaro V, Vari S, Cognetti F and Ciuffreda L: PTEN: Multiple functions in human malignant tumors. Front Oncol. 5:242015. View Article : Google Scholar : PubMed/NCBI
24	Koriyama Y, Kamiya M, Arai K, Sugitani K, Ogai K and Kato S: Nipradilol promotes axon regeneration through S-nitrosylation of PTEN in retinal ganglion cells. Adv Exp Med Biol. 801:751–757. 2014. View Article : Google Scholar : PubMed/NCBI
25	Li R, Jin Y, Li Q, Sun X, Zhu H and Cui H: MiR-93-5p targeting PTEN regulates the NMDA-induced autophagy of retinal ganglion cells via AKT/mTOR pathway in glaucoma. Biomed Pharmacother. 100:1–7. 2018. View Article : Google Scholar : PubMed/NCBI
26	Leslie NR, Bennett D, Lindsay YE, Stewart H, Gray A and Downes CP: Redox regulation of PI 3-kinase signalling via inactivation of PTEN. EMBO J. 22:5501–5510. 2003. View Article : Google Scholar : PubMed/NCBI
27	Yu AL, Fuchshofer R, Kampik A and Welge-Lüssen U: Effects of oxidative stress in trabecular meshwork cells are reduced by prostaglandin analogues. Invest Ophthalmol Vis Sci. 49:4872–4880. 2008. View Article : Google Scholar : PubMed/NCBI
28	Saccà SC, Pulliero A and Izzotti A: The dysfunction of the trabecular meshwork during glaucoma course. J Cell Physiol. 230:510–525. 2015. View Article : Google Scholar : PubMed/NCBI
29	Wang Y, Li F and Wang S: MicroRNA-93 is overexpressed and induces apoptosis in glaucoma trabecular meshwork cells. Mol Med Rep. 14:5746–5750. 2016. View Article : Google Scholar : PubMed/NCBI
30	Yuan C, Xu M, Rong R, Mei Y, Cai W, Li L, Xue Y, Zhu B, Sun K and Han L: miR-200c regulates endothelin-1 induced PASMCs abnormal proliferation and apoptosis. IUBMB Life. 69:877–886. 2017. View Article : Google Scholar : PubMed/NCBI
31	Li MF, Guan H and Zhang DD: Effect of overexpression of PTEN on apoptosis of liver cancer cells. Genet Mol Res. 15:2016.
32	Baleriola J, García-Feijoo J, Martínez-de-la-Casa JM, Fernández-Cruz A, de la Rosa EJ and Fernández-Durango R: Apoptosis in the trabecular meshwork of glaucomatous patients. Mol Vis. 14:1513–1516. 2008.PubMed/NCBI
33	Steckley D, Karajgikar M, Dale LB, Fuerth B, Swan P, Drummond-Main C, Poulter MO, Ferguson SS, Strasser A and Cregan SP: Puma is a dominant regulator of oxidative stress induced Bax activation and neuronal apoptosis. J Neurosci. 27:12989–12999. 2007. View Article : Google Scholar : PubMed/NCBI
34	Awai-Kasaoka N, Inoue T, Kameda T, Fujimoto T, Inoue-Mochita M and Tanihara H: Oxidative stress response signaling pathways in trabecular meshwork cells and their effects on cell viability. Mol Vis. 19:1332–1340. 2013.PubMed/NCBI
35	Han Z, Chen F, Ge X, Tan J, Lei P and Zhang J: miR-21 alleviated apoptosis of cortical neurons through promoting PTEN-Akt signaling pathway in vitro after experimental traumatic brain injury. Brain Res. 1582:12–20. 2014. View Article : Google Scholar : PubMed/NCBI
36	Chalhoub N and Baker SJ: PTEN and the PI3-kinase pathway in cancer. Annu Rev Pathol. 4:127–150. 2009. View Article : Google Scholar : PubMed/NCBI
37	Chen JS, Wang Q, Fu XH, Huang XH, Chen XL, Cao LQ, Chen LZ, Tan HX, Li W, Bi J and Zhang LJ: Involvement of PI3K/PTEN/AKT/mTOR pathway in invasion and metastasis in hepatocellular carcinoma: Association with MMP-9. Hepatol Res. 39:177–186. 2009. View Article : Google Scholar : PubMed/NCBI