MicroRNA‑145‑5p inhibits osteosarcoma cell proliferation by targeting E2F transcription factor 3

Li,Hao; Pan,Runsang; Lu,Qiaoying; Ren,Chong; Sun,Junkang; Wu,Huaping; Wen,Jing; Chen,Houping

doi:10.3892/ijmm.2020.4504

MicroRNA‑145‑5p inhibits osteosarcoma cell proliferation by targeting E2F transcription factor 3

Authors:
- Hao Li
- Runsang Pan
- Qiaoying Lu
- Chong Ren
- Junkang Sun
- Huaping Wu
- Jing Wen
- Houping Chen
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Affiliations: Department of Orthopedics, Guiyang Maternal and Child Health-Care Hospital, Guiyang, Guizhou 550000, P.R. China, Department of Pathology, Renmin Hospital of Guizhou, Guiyang, Guizhou 550000, P.R. China
Published online on: February 17, 2020 https://doi.org/10.3892/ijmm.2020.4504
Pages: 1317-1326
Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Osteosarcoma is a common type of bone tumor that primarily occurs in children and young adults. MicroRNA (miRNA/miR) dysregulation is associated with the progression of osteosarcoma; therefore, the aim of the present study was to investigate the biological functions and molecular mechanisms of miR‑145‑5p in osteosarcoma. The expression of miR‑145‑5p in osteosarcoma tissues and cell lines was quantified using reverse transcription‑quantitative PCR (RT‑qPCR). The effect of miR‑145‑5p on the proliferation of osteosarcoma cells was detected using Cell Counting Kit‑8 and colony formation assays, as well as cell cycle distribution analysis. The effect of miR‑145‑5p on tumor growth was further investigated in vivo using a subcutaneous tumor model in nude mice. The interaction between miR‑145‑5p and E2F transcription factor 3 (E2F3) was determined using bioinformatics analysis, a luciferase assay, RT‑qPCR and western blotting. The results revealed that miR‑145‑5p expression was decreased in osteosarcoma cell lines and tissues compared with the corresponding normal controls. Increased miR‑145‑5p expression inhibited the proliferation and colony formation ability of osteosarcoma cells, and induced G1 phase arrest. Furthermore, mice injected with tumor cells overexpressing miR‑145‑5p exhibited smaller tumors than those in the control group. Further investigation revealed that miR‑145‑5p binds to and decreases the expression of E2F3. In addition, the mRNA levels of E2F3 were negatively associated with miR‑145‑5p in osteosarcoma tissues, and increasing E2F3 expression abrogated the inhibitory effects of miR‑145‑5p on osteosarcoma cells. Collectively, the results obtained in the present study suggest that miR‑145‑5p may suppress the progression of osteosarcoma, and may serve as a useful biomarker for the diagnosis of osteosarcoma, as well as a therapeutic target.

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1	Fox E, Levin K, Zhu Y, Segers B, Balamuth N, Womer R, Bagatell R and Balis F: Pantoprazole, an inhibitor of the organic cation transporter 2, does not ameliorate cisplatin-related ototoxicity or nephrotoxicity in children and adolescents with newly diagnosed osteosarcoma treated with methotrexate, doxorubicin, and cisplatin. Oncologist. 23:e762–e779. 2018. View Article : Google Scholar
2	Endicott AA, Morimoto LM, Kline CN, Wiemels JL, Metayer C and Walsh KM: Perinatal factors associated with clinical presentation of osteosarcoma in children and adolescents. Pediatr Blood Cancer. 64:e263492017. View Article : Google Scholar
3	Kebudi R, Ozger H, Kizilocak H, Bay SB and Bilgic B: Osteosarcoma after hematopoietic stem cell transplantation in children and adolescents: Case report and review of the literature. Pediatr Blood Cancer. 63:1664–1666. 2016. View Article : Google Scholar : PubMed/NCBI
4	Lee JA, Paik EK, Seo J, Kim DH, Lim JS, Yoo JY and Kim MS: Radiotherapy and gemcitabine-docetaxel chemotherapy in children and adolescents with unresectable recurrent or refractory osteosarcoma. Jpn J Clin Oncol. 46:138–143. 2016.
5	Han Q, Li C, Cao Y, Bao J, Li K, Song R, Chen X, Li J and Wu X: CBX2 is a functional target of miRNA let-7a and acts as a tumor promoter in osteosarcoma. Cancer Med. 8:3981–3991. 2019.PubMed/NCBI
6	Huang C, Wang Q, Ma S, Sun Y, Vadamootoo AS and Jin C: A four serum-miRNA panel serves as a potential diagnostic biomarker of osteosarcoma. Int J Clin Oncol. 24:976–982. 2019. View Article : Google Scholar : PubMed/NCBI
7	Wang H, Zhao F, Cai S and Pu Y: MiR-193a regulates chemoresistance of human osteosarcoma cells via repression of IRS2. J Bone Oncol. 17:1002412019. View Article : Google Scholar : PubMed/NCBI
8	Liu XG, Xu J, Li F, Li MJ and Hu T: Down-regulation of miR-377 contributes to cisplatin resistance by targeting XIAP in osteosarcoma. Eur Rev Med Pharmacol Sci. 22:1249–1257. 2018.PubMed/NCBI
9	Hu XH, Dai J, Shang HL, Zhao ZX and Hao YD: MiR-1285-3p is a potential prognostic marker in human osteosarcoma and functions as a tumor suppressor by targeting YAP1. Cancer Biomark. 25:1–10. 2019. View Article : Google Scholar : PubMed/NCBI
10	Chu Y, Hu X, Wang G, Wang Z and Wang Y: Downregulation of miR-136 promotes the progression of osteosarcoma and is associated with the prognosis of patients with osteosarcoma. Oncol Lett. 17:5210–5218. 2019.PubMed/NCBI
11	Wang JY, Lu SB and Wang JQ: MiR-628-5p promotes the growth and migration of osteosarcoma by targeting IFI44L. Biochem Cell Biol. Apr 24–2019.(Epub ahead of print).
12	Ouyang L, Liu P, Yang S, Ye S, Xu W and Liu X: A three-plasma miRNA signature serves as novel biomarkers for osteosarcoma. Med Oncol. 30:3402013. View Article : Google Scholar
13	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
14	Bertoli C, Skotheim JM and de Bruin RA: Control of cell cycle transcription during G1 and S phases. Nat Rev Mol Cell Biol. 14:518–528. 2013. View Article : Google Scholar : PubMed/NCBI
15	Crncec A and Hochegger H: Triggering mitosis. FEBS Lett. 593:2868–2888. 2019. View Article : Google Scholar : PubMed/NCBI
16	Schwentner R, Papamarkou T, Kauer MO, Stathopoulos V, Yang F, Bilke S, Meltzer PS, Girolami M and Kovar H: EWS-FLI1 employs an E2F switch to drive target gene expression. Nucleic Acids Res. 43:2780–2789. 2015. View Article : Google Scholar : PubMed/NCBI
17	Zhang H, Jiang M, Liu Q, Han Z, Zhao Y and Ji S: MiR-145-5p inhibits the proliferation and migration of bladder cancer cells by targeting TAGLN2. Oncol Lett. 16:6355–6360. 2018.PubMed/NCBI
18	Zhuang S, Liu F and Wu P: Upregulation of long noncoding RNA TUG1 contributes to the development of laryngocarcinoma by targeting miR-145-5p/ROCK1 axis. J Cell Biochem. 120:13392–13402. 2019. View Article : Google Scholar : PubMed/NCBI
19	Goeppert B, Truckenmueller F, Ori A, Fritz V, Albrecht T, Fraas A, Scherer D, Silos RG, Sticht C, Gretz N, et al: Profiling of gallbladder carcinoma reveals distinct miRNA profiles and activation of STAT1 by the tumor suppressive miRNA-145-5p. Sci Rep. 9:47962019. View Article : Google Scholar : PubMed/NCBI
20	Wang B, Dong W and Li X: MiR-145-5p acts as a novel tumor suppressor in hepatocellular carcinoma through targeting RAB18. Technol Cancer Res Treat. 18:1533033819850189. 2019. View Article : Google Scholar
21	Tang W, Zhang X, Tan W, Gao J, Pan L, Ye X, Chen L and Zheng W: MiR-145-5p suppresses breast cancer progression by inhibiting SOX2. J Surg Res. 236:278–287. 2019. View Article : Google Scholar : PubMed/NCBI
22	Jian M, Du Q, Zhu D, Mao Z, Wang X, Feng Y, Xiao Z, Wang H and Zhu Y: Tumor suppressor miR-145-5p sensitizes prolactinoma to bromocriptine by downregulating TPT1. J Endocrinol Invest. 42:639–652. 2019. View Article : Google Scholar
23	Zhou T, Chen S and Mao X: MiR-145-5p affects the differentiation of gastric cancer by targeting KLF5 directly. J Cell Physiol. 234:7634–7644. 2019. View Article : Google Scholar
24	Wu Z, Huang W, Wang X, Wang T, Chen Y, Chen B, Liu R, Bai P and Xing J: Circular RNA CEP128 acts as a sponge of miR-145-5p in promoting the bladder cancer progression via regulating SOX11. Mol Med. 24:402018. View Article : Google Scholar : PubMed/NCBI
25	Shen ZG, Liu XZ, Chen CX and Lu JM: Knockdown of E2F3 inhibits proliferation, migration, and invasion and increases apoptosis in glioma cells. Oncol Res. 25:1555–1566. 2017. View Article : Google Scholar : PubMed/NCBI
26	Dong D, Gong Y, Zhang D, Bao H and Gu G: MiR-874 suppresses the proliferation and metastasis of osteosarcoma by targeting E2F3. Tumour Biol. 37:6447–6455. 2016. View Article : Google Scholar
27	Jin Y, Wei J, Xu S, Guan F, Yin L and Zhu H: MiR210-3p regulates cell growth and affects cisplatin sensitivity in human ovarian cancer cells via targeting E2F3. Mol Med Rep. 19:4946–4954. 2019.PubMed/NCBI
28	Liu J, Si L and Tian H: MicroRNA-148a inhibits cell proliferation and cell cycle progression in lung adenocarcinoma via directly targeting transcription factor E2F3. Exp Ther Med. 16:5400–5409. 2018.PubMed/NCBI
29	Feng Z, Peng C, Li D, Zhang D, Li X, Cui F, Chen Y and He Q: E2F3 promotes cancer growth and is overexpressed through copy number variation in human melanoma. Onco Targets Ther. 11:5303–5313. 2018. View Article : Google Scholar : PubMed/NCBI
30	Han R, Chen X, Li Y, Zhang S, Li R and Lu L: MicroRNA-34a suppresses aggressiveness of hepatocellular carcinoma by modulating E2F1, E2F3, and Caspase-3. Cancer Manag Res. 11:2963–2976. 2019. View Article : Google Scholar :
31	Sun FB, Lin Y, Li SJ, Gao J, Han B and Zhang CS: MiR-210 knockdown promotes the development of pancreatic cancer via upregulating E2F3 expression. Eur Rev Med Pharmacol Sci. 22:8640–8648. 2018.PubMed/NCBI
32	Yang J, Zhang HF and Qin CF: MicroRNA-217 functions as a prognosis predictor and inhibits pancreatic cancer cell proliferation and invasion via targeting E2F3. Eur Rev Med Pharmacol Sci. 21:4050–4057. 2017.PubMed/NCBI