Emerging role of microRNA-27a in human malignant glioma cell survival via targeting of prohibitin

Chen,Wei; Qi,Jun; Bao,Gang; Wang,Tuo; Du,Chang‑Wang; Wang,Mao‑De

doi:10.3892/mmr.2015.3475

Emerging role of microRNA-27a in human malignant glioma cell survival via targeting of prohibitin

Authors:
- Wei Chen
- Jun Qi
- Gang Bao
- Tuo Wang
- Chang‑Wang Du
- Mao‑De Wang
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Affiliations: Department of Neurosurgery, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China, Institute of Transfusion Research, Shaanxi Blood Center, Xi'an, Shaanxi 710061, P.R. China
Published online on: March 12, 2015 https://doi.org/10.3892/mmr.2015.3475
Pages: 1515-1523

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Abstract

MicroRNAs (miRs) function as oncogenes and tumor suppressors, and have roles in most cellular processes. To date, the possible role of miR‑27a, which is part of the miR‑23a/27a/24‑2 cluster, in malignant gliomas has remained elusive. Therefore, the present study aimed to explore the role of miR‑27a in glioma and its potential target. Through transfection with miR‑27a inhibitor or oligonucleotide mimics, the impact of miR‑27a silencing or overexpression on the growth, apoptosis, cell cycle and invasiveness of U251 and U87MG cells was examined in vitro. The present study initially identified the potential target of miR‑27a in glioma cells through a bioinformatics analysis, which was used for screening of the literature on the proteomics of gliomas. Prohibitin (PHB) was then confirmed as a target by absolute luciferase reporter assays, quantitative real‑time polymerase chain reaction and western blot analysis. Treatment with miR‑27a mimics oligonucleotides suppressed U251 cell proliferation, promoted apoptosis by inducing G2/M phase arrest, and impaired the invasive potential of U251 cells in vitro. In addition, miR‑27a expression was downregulated in glioma tissues. A PHB‑3'‑untranslated region luciferase reporter assay confirmed PHB as a direct target gene of miR‑27a. PHB mRNA expression was reversely correlated with levels of miR‑27a in U251 cells. Overexpression of miR‑27a in U251 cells at 72 h and 96 h post‑transfection with miR‑27a mimics significantly decreased PHB protein expression by 17.2% and 43.9%, respectively. In conclusion, miR‑27a was shown to be a significant tumor suppressor by targeting and decreasing PHB protein expression in glioma U251 cells. miR‑27a targeting of PHB may be a novel potential therapeutic strategy for glioma.

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1	Wen PY and Kesari S: Malignant gliomas in adults. N Engl J Med. 359:492–507. 2008. View Article : Google Scholar : PubMed/NCBI
2	Gabayan AJ, Green SB, Sanan A, et al: GliaSite brachytherapy for treatment of recurrent malignant gliomas: a retrospective multi-institutional analysis. Neurosurgery. 58:701–709. 2006. View Article : Google Scholar : PubMed/NCBI
3	Bartel DP: MicroRNAs: target recognition and regulatory functions. Cell. 136:215–233. 2009. View Article : Google Scholar : PubMed/NCBI
4	Cho WC: OncomiRs: the discovery and progress of microRNAs in cancers. Mol Cancer. 6:602007. View Article : Google Scholar : PubMed/NCBI
5	Anderson E, Grant R, Lewis SC and Whittle IR: Randomized Phase III controlled trials of therapy in malignant glioma: where are we after 40 years? Br J Neurosurg. 22:339–349. 2008. View Article : Google Scholar : PubMed/NCBI
6	Auffinger B, Thaci B, Ahmed A, et al: MicroRNA targeting as a therapeutic strategy against glioma. Curr Mol Med. 13:535–542. 2013. View Article : Google Scholar
7	Chhabra R, Adlakha YK, Hariharan M, Scaria V and Saini N: Upregulation of miR-23a-27a-24-2 cluster induces caspase-dependent and -independent apoptosis in human embryonic kidney cells. PLoS One. 4:e58482009. View Article : Google Scholar : PubMed/NCBI
8	Chhabra R, Dubey R and Saini N: Cooperative and individualistic functions of the microRNAs in the miR-23a~27a~24-2 cluster and its implication in human diseases. Mol Cancer. 9:2322010. View Article : Google Scholar : PubMed/NCBI
9	Feng DD, Zhang H, Zhang P, et al: Down-regulated miR-331-5p and miR-27a are associated with chemotherapy resistance and relapse in leukaemia. J Cell Mol Med. 15:2164–2175. 2011. View Article : Google Scholar
10	Ballabio E, Armesto M, Breeze CE, et al: Bortezomib action in multiple myeloma: microRNA-mediated synergy (and miR-27a/CDK5 driven sensitivity)? Blood Cancer J. 2:e832012. View Article : Google Scholar : PubMed/NCBI
11	Chow TF, Youssef YM, Lianidou E, et al: Differential expression profiling of microRNAs and their potential involvement in renal cell carcinoma pathogenesis. Clin Biochem. 43:150–158. 2010. View Article : Google Scholar
12	Guttilla IK and White BA: Coordinate regulation of FOXO1 by miR-27a, miR-96 and miR-182 in breast cancer cells. J Biol Chem. 284:23204–23216. 2009. View Article : Google Scholar : PubMed/NCBI
13	Huang S, He X, Ding J, et al: Upregulation of miR-23a approximately 27a approximately 24 decreases transforming growth factor-beta-induced tumor-suppressive activities in human hepatocellular carcinoma cells. Int J Cancer. 123:972–978. 2008. View Article : Google Scholar : PubMed/NCBI
14	Prueitt RL, Yi M, Hudson RS, et al: Expression of microRNAs and protein-coding genes associated with perineural invasion in prostate cancer. Prostate. 68:1152–1164. 2008. View Article : Google Scholar : PubMed/NCBI
15	Saumet A, Vetter G, Bouttier M, et al: Transcriptional repression of microRNA genes by PML-RARA increases expression of key cancer proteins in acute promyelocytic leukemia. Blood. 113:412–421. 2009. View Article : Google Scholar
16	Xi Y, Shalgi R, Fodstad O, Pilpel Y and Ju J: Differentially regulated micro-RNAs and actively translated messenger RNA transcripts by tumor suppressor p53 in colon cancer. Clin Cancer Res. 12(7 Pt 1): 2014–2024. 2006. View Article : Google Scholar : PubMed/NCBI
17	Wang W, Cheng B, Miao L, Mei Y and Wu M: Mutant p53-R273H gains new function in sustained activation of EGFR signaling via suppressing miR-27a expression. Cell Death Dis. 4:e5742013. View Article : Google Scholar : PubMed/NCBI
18	Deighton RF, McGregor R, Kemp J, McCulloch J and Whittle IR: Glioma pathophysiology: insights emerging from proteomics. Brain Pathol. 20:691–703. 2010. View Article : Google Scholar : PubMed/NCBI
19	Chumbalkar VC, Subhashini C, Dhople VM, et al: Differential protein expression in human gliomas and molecular insights. Proteomics. 5:1167–1177. 2005. View Article : Google Scholar : PubMed/NCBI
20	Hiratsuka M, Inoue T, Toda T, et al: Proteomics-based identification of differentially expressed genes in human gliomas: down-regulation of SIRT2 gene. Biochem Biophys Res Commun. 309:558–566. 2003. View Article : Google Scholar : PubMed/NCBI
21	Iwadate Y, Sakaida T, Hiwasa T, et al: Molecular classification and survival prediction in human gliomas based on proteome analysis. Cancer Res. 64:2496–2501. 2004. View Article : Google Scholar : PubMed/NCBI
22	Zhou JQ, Wang JT, Liu QH, Guo Xb, Zhou J and Song LJ: Proteomic profiling and identification of malignant grade related proteins in human brain astrocytoma. Chin J Neuromed. 11:780–783. 2012.
23	Mishra S, Murphy LC and Murphy LJ: The Prohibitins: emerging roles in diverse functions. J Cell Mol Med. 10:353–363. 2006. View Article : Google Scholar : PubMed/NCBI
24	Mishra S, Murphy LC, Nyomba BL and Murphy LJ: Prohibitin: a potential target for new therapeutics. Trends Mol Med. 11:192–197. 2005. View Article : Google Scholar : PubMed/NCBI
25	Liu T, Tang H, Lang Y, Liu M and Li X: MicroRNA-27a functions as an oncogene in gastric adenocarcinoma by targeting prohibitin. Cancer Lett. 273:233–242. 2009. View Article : Google Scholar
26	Mertens-Talcott SU, Chintharlapalli S, Li X and Safe S: The oncogenic microRNA-27a targets genes that regulate specificity protein transcription factors and the G2-M checkpoint in MDA-MB-231 breast cancer cells. Cancer Res. 67:11001–11011. 2007. View Article : Google Scholar : PubMed/NCBI
27	Scott GK, Mattie MD, Berger CE, Benz SC and Benz CC: Rapid alteration of microRNA levels by histone deacetylase inhibition. Cancer Res. 66:1277–1281. 2006. View Article : Google Scholar : PubMed/NCBI
28	Lerner M, Lundgren J, Akhoondi S, et al: MiRNA-27a controls FBW7/hCDC4-dependent cyclin E degradation and cell cycle progression. Cell Cycle. 10:2172–2183. 2011. View Article : Google Scholar : PubMed/NCBI
29	Belair C, Darfeuille F and Staedel C: Helicobacter pylori and gastric cancer: possible role of microRNAs in this intimate relationship. Clin Microbiol Infect. 15:806–812. 2009. View Article : Google Scholar : PubMed/NCBI
30	Chintharlapalli S, Papineni S, Abdelrahim M, et al: Oncogenic microRNA-27a is a target for anticancer agent methyl 2-cyano-3,11-dioxo-18beta-olean-1,12-dien-30-oate in colon cancer cells. Int J Cancer. 125:1965–1974. 2009. View Article : Google Scholar : PubMed/NCBI
31	Volinia S, Calin GA, Liu CG, et al: A microRNA expression signature of human solid tumors defines cancer gene targets. Proc Natl Acad Sci USA. 103:2257–2261. 2006. View Article : Google Scholar : PubMed/NCBI
32	Schultz J, Lorenz P, Gross G, Ibrahim S and Kunz M: MicroRNA let-7b targets important cell cycle molecules in malignant melanoma cells and interferes with anchorage-independent growth. Cell Res. 18:549–557. 2008. View Article : Google Scholar : PubMed/NCBI
33	Kozaki K, Imoto I, Mogi S, Omura K and Inazawa J: Exploration of tumor-suppressive microRNAs silenced by DNA hypermethylation in oral cancer. Cancer Res. 68:2094–2105. 2008. View Article : Google Scholar : PubMed/NCBI
34	Scheibner KA, Teaboldt B, Hauer MC, et al: MiR-27a functions as a tumor suppressor in acute leukemia by regulating 14-3-3theta. PLoS One. 7:e508952012. View Article : Google Scholar
35	Zhou TB and Qin YH: Signaling pathways of prohibitin and its role in diseases. J Recept Signal Transduct Res. 33:28–36. 2013. View Article : Google Scholar : PubMed/NCBI
36	Nijtmans LG, de Jong L, Artal Sanz M, et al: Prohibitins act as a membrane-bound chaperone for the stabilization of mitochondrial proteins. EMBO J. 19:2444–2451. 2000. View Article : Google Scholar : PubMed/NCBI
37	Chowdhury I, Thompson WE, Welch C, Thomas K and Matthews R: Prohibitin (PHB) inhibits apoptosis in rat granulosa cells (GCs) through the extracellular signal-regulated kinase 1/2 (ERK1/2) and the Bcl family of proteins. Apoptosis. 18:1513–1525. 2013. View Article : Google Scholar : PubMed/NCBI
38	Kasashima K, Ohta E, Kagawa Y and Endo H: Mitochondrial functions and estrogen receptor-dependent nuclear translocation of pleiotropic human prohibitin 2. J Biol Chem. 281:36401–36410. 2006. View Article : Google Scholar : PubMed/NCBI
39	Merkwirth C, Dargazanli S, Tatsuta T, et al: Prohibitins control cell proliferation and apoptosis by regulating OPA1-dependent cristae morphogenesis in mitochondria. Genes Dev. 22:476–488. 2008. View Article : Google Scholar : PubMed/NCBI
40	Ross JA, Nagy ZS and Kirken RA: The PHB1/2 phosphocomplex is required for mitochondrial homeostasis and survival of human T cells. J Biol Chem. 283:4699–4713. 2008. View Article : Google Scholar
41	Rajalingam K, Wunder C, Brinkmann V, et al: Prohibitin is required for Ras-induced Raf-MEK-ERK activation and epithelial cell migration. Nat Cell Biol. 7:837–843. 2005. View Article : Google Scholar : PubMed/NCBI
42	Chiu CF, Ho MY, Peng JM, et al: Raf activation by Ras and promotion of cellular metastasis require phosphorylation of prohibitin in the raft domain of the plasma membrane. Oncogene. 32:777–787. 2013. View Article : Google Scholar
43	Muraguchi T, Kawawa A and Kubota S: Prohibitin protects against hypoxia-induced H9c2 cardiomyocyte cell death. Biomed Res. 31:113–122. 2010. View Article : Google Scholar : PubMed/NCBI
44	Fusaro G, Wang S and Chellappan S: Differential regulation of Rb family proteins and prohibitin during camptothecin-induced apoptosis. Oncogene. 21:4539–4548. 2002. View Article : Google Scholar : PubMed/NCBI
45	Phillips AC and Vousden KH: E2F-1 induced apoptosis. Apoptosis. 6:173–182. 2001. View Article : Google Scholar : PubMed/NCBI
46	Fusaro G, Dasgupta P, Rastogi S, Joshi B and Chellappan S: Prohibitin induces the transcriptional activity of p53 and is exported from the nucleus upon apoptotic signaling. J Biol Chem. 278:47853–47861. 2003. View Article : Google Scholar : PubMed/NCBI
47	Clark MJ, Homer N, O’Connor BD, et al: U87MG decoded: the genomic sequence of a cytogenetically aberrant human cancer cell line. PLoS Genet. 6:e10008322010. View Article : Google Scholar : PubMed/NCBI