MicroRNA‑20a promotes the proliferation and cell cycle of human osteosarcoma cells by suppressing early growth response 2 expression

Zhuo,Wenkun; Ge,Weiming; Meng,Guolin; Jia,Shuaijun; Zhou,Xiang; Liu,Jian

doi:10.3892/mmr.2015.4098

MicroRNA‑20a promotes the proliferation and cell cycle of human osteosarcoma cells by suppressing early growth response 2 expression

Authors:
- Wenkun Zhuo
- Weiming Ge
- Guolin Meng
- Shuaijun Jia
- Xiang Zhou
- Jian Liu
View Affiliations

Affiliations: Department of Orthopedics and Traumatology, Institute of Orthopedics and Traumatology, Xijing Hospital, The Fourth Military Medical University, Xi'an, Shanxi 710000, P.R. China, Department of Orthopedics and Traumatology, The PLA Fifth Thirty Four Hospital, Luoyang, Henan 471000, P.R. China, Department of Orthopedics, Shanxi Hospital of Chinese Armed Police Forces, Xi'an, Shanxi 710000, P.R. China
Published online on: July 20, 2015 https://doi.org/10.3892/mmr.2015.4098
Pages: 4989-4994
Copyright: © Zhuo 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

MicroRNAs (miRNAs) are crucial in cancer development. However, the underlying mechanisms of miRNAs in osteosarcoma (OS) remain largely uncharacterized. The present study investigated the role of miR‑20a in OS cell proliferation. It was determined that miR‑20a expression is markedly upregulated in OS tissues and cells compared with the matched adjacent normal tissues and h‑FOB human osteoblast cell lines. Ectopic expression of miR‑20a promoted the proliferation and anchorage‑independent growth of OS cells, whereas inhibition of miR‑20a reduced this effect. Bioinformatics analysis further revealed early growth response 2 (EGR2), as a potential target of miR‑20a. Data from luciferase reporter assays showed that miR‑20a directly binds to the 3'‑untranslated region (3'‑UTR) of EGR2 mRNA and represses expression at the transcriptional and translational levels. In functional assays, miR‑20a promoted OS cell proliferation and the cell cycle, which could be suppressed by an inhibitor of miR‑20a. In conclusion, the data provide compelling evidence that miR‑20a functions as an onco‑miRNA, which is important in promoting cell proliferation in OS, and its oncogenic effect is mediated primarily through direct suppression of EGR2 expression.

View Figures

View References

1	Ottaviani G and Jaffe N: The epidemiology of osteosarcoma. Cancer Treat Res. 152:3–13. 2009. View Article : Google Scholar
2	Ahmed AA, Zia H and Wagner L: Therapy resistance mechanisms in Ewing's sarcoma family tumors. Cancer Chemother Pharmacol. 73:657–663. 2014. View Article : Google Scholar : PubMed/NCBI
3	Constantinidou A, Pollack S, Loggers E, Rodler E and Jones RL: The evolution of systemic therapy in sarcoma. Expert Rev Anticancer Ther. 13:211–223. 2013. View Article : Google Scholar : PubMed/NCBI
4	Miao J, Wu S, Peng Z, Tania M and Zhang C: MicroRNAs in osteosarcoma: Diagnostic and therapeutic aspects. Tumour Biol. 34:2093–2098. 2013. View Article : Google Scholar : PubMed/NCBI
5	Esquela-Kerscher A and Slack FJ: Oncomirs - microRNAs with a role in cancer. Nat Rev Cancer. 6:259–269. 2006. View Article : Google Scholar : PubMed/NCBI
6	Calin GA and Croce CM: MicroRNA signatures in human cancers. Nat Rev Cancer. 6:857–866. 2006. View Article : Google Scholar : PubMed/NCBI
7	Sun K and Lai EC: Adult-specific functions of animal microRNAs. Nat Rev Genet. 14:535–548. 2013. View Article : Google Scholar : PubMed/NCBI
8	Xiong Y, Zhang L and Kebebew E: MiR-20a is upregulated in anaplastic thyroid cancer and targets LIMK1. PLoS One. 9:e961032014. View Article : Google Scholar : PubMed/NCBI
9	Zhao S, Yao D, Chen J and Ding N: Circulating miRNA-20a and miRNA-203 for screening lymph node metastasis in early stage cervical cancer. Genet Test Mol Biomarkers. 17:631–636. 2013. View Article : Google Scholar : PubMed/NCBI
10	Chang Y, Liu C, Yang J, Liu G, Feng F, Tang J, Hu L, Li L, Jiang F, Chen C, et al: MiR-20a triggers metastasis of gallbladder carcinoma. J Hepatol. 59:518–527. 2013. View Article : Google Scholar : PubMed/NCBI
11	Joseph LJ, Le Beau MM, Jamieson GA Jr, Acharya S, Shows TB, Rowley JD and Sukhatme VP: Molecular cloning, sequencing, and mapping of EGR2, a human early growth response gene encoding a protein with “zinc-binding finger” structure. Proc Natl Acad Sci USA. 85:7164–7168. 1988. View Article : Google Scholar
12	Li X, Zhang Z, Yu M, Li L, Du G, Xiao W and Yang H: Involvement of miR-20a in promoting gastric cancer progression by targeting early growth response 2 (EGR2). Int J Mol Sci. 14:16226–16239. 2013. View Article : Google Scholar : PubMed/NCBI
13	Sumitomo S, Fujio K, Okamura T and Yamamoto K: Egr2 and Egr3 are the unique regulators for systemic autoimmunity. JAK-STAT. 2:e239522013. View Article : Google Scholar : PubMed/NCBI
14	Yu Z, Wang C, Wang M, Li Z, Casimiro MC, Liu M, Wu K, Whittle J, Ju X, Hyslop T, et al: A cyclin D1/microRNA 17/20 regulatory feedback loop in control of breast cancer cell proliferation. J Cell Biol. 182:509–517. 2008. View Article : Google Scholar : PubMed/NCBI
15	Wang Z, Liu M, Zhu H, Zhang W, He S, Hu C, Quan L, Bai J and Xu N: Suppression of p21 by c-Myc through members of miR-17 family at the post-transcriptional level. Int J Oncol. 37:1315–1321. 2010.PubMed/NCBI
16	Arabi L, Gsponer JR, Smida J, Nathrath M, Perrina V, Jundt G, Ruiz C, Quagliata L and Baumhoer D: Upregulation of the miR-17-92 cluster and its two paraloga in osteosarcoma - reasons and consequences. Genes Cancer. 5:56–63. 2014.PubMed/NCBI
17	Tian Q, Jia J, Ling S, Liu Y, Yang S and Shao Z: A causal role for circulating miR-34b in osteosarcoma. Eur J Surg Oncol. 40:67–72. 2014. View Article : Google Scholar
18	Xu JQ, Liu P, Si MJ and Ding XY: MicroRNA-126 inhibits osteosarcoma cells proliferation by targeting Sirt1. Tumour Biol. 34:3871–3877. 2013. View Article : Google Scholar : PubMed/NCBI
19	Cao ZQ, Shen Z and Huang WY: MicroRNA-802 promotes osteosarcoma cell proliferation by targeting p27. Asian Pac J Cancer Prev. 14:7081–7084. 2013. View Article : Google Scholar
20	Tang M, Lin L, Cai H, Tang J and Zhou Z: MicroRNA-145 downregulation associates with advanced tumor progression and poor prognosis in patients suffering osteosarcoma. Onco Targets Ther. 6:833–838. 2013.PubMed/NCBI
21	van Kouwenhove M, Kedde M and Agami R: MicroRNA regulation by RNA-binding proteins and its implications for cancer. Nat Rev Cancer. 11:644–656. 2011. View Article : Google Scholar : PubMed/NCBI
22	Unoki M and Nakamura Y: Growth-suppressive effects of BPOZ and EGR2, two genes involved in the PTEN signaling pathway. Oncogene. 20:4457–4465. 2001. View Article : Google Scholar : PubMed/NCBI
23	Baldin V, Lukas J, Marcote MJ, Pagano M and Draetta G: Cyclin D1 is a nuclear protein required for cell cycle progression in G1. Genes Dev. 7:812–821. 1993. View Article : Google Scholar : PubMed/NCBI
24	Masamha CP and Benbrook DM: Cyclin D1 degradation is sufficient to induce G1 cell cycle arrest despite constitutive expression of cyclin E2 in ovarian cancer cells. Cancer Res. 69:6565–6572. 2009. View Article : Google Scholar : PubMed/NCBI
25	Yang M, Zhong J, Zhao M, Wang J, Gu Y, Yuan X, Sang J and Huang C: Overexpression of nuclear apoptosis-inducing factor 1 altered the proteomic profile of human gastric cancer cell MKN45 and induced cell cycle arrest at G1/S phase. PLoS One. 9:e1002162014. View Article : Google Scholar : PubMed/NCBI