FOXN3 is downregulated in osteosarcoma and transcriptionally regulates SIRT6, and suppresses migration and invasion in osteosarcoma

Xue,Wentao; Ma,Linjie; Wang,Zhiqian; Zhang,Weidong; Zhang,Xiugong

doi:10.3892/or.2018.6878

FOXN3 is downregulated in osteosarcoma and transcriptionally regulates SIRT6, and suppresses migration and invasion in osteosarcoma

Authors:
- Wentao Xue
- Linjie Ma
- Zhiqian Wang
- Weidong Zhang
- Xiugong Zhang
View Affiliations

Affiliations: Department of Orthopedics, Yidu Central Hospital of Weifang, Weifang, Shandong 262500, P.R. China, Department of Orthopedics, Yantai Hospital of Traditional Chinese Medicine, Yantai, Shandong 264000, P.R. China, Department of Orthopedics, Qingdao Central Hospital, Qingdao, Shandong 266042, P.R. China
Published online on: November 21, 2018 https://doi.org/10.3892/or.2018.6878
Pages: 1404-1414

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

Forkhead box N3 (FOXN3) has been reported to be downregulated in numerous cancers, including laryngeal, oral squamous cell and hepatocellular carcinomas, and diffuse large B‑cell lymphoma. FOXN3 was proposed to serve as a tumor suppressor; however, the function of FOXN3 in osteosarcoma (OS) remains unknown. The present study suggested that FOXN3 was notably downregulated in OS tissues compared with in adjacent normal tissues, and the expression of FOXN3 was negatively correlated with tumor size, metastasis and tumor, node and metastasis stage. Additionally, low expression levels of FOXN3 predicted a poor prognosis of patients with OS. Additionally, the present study revealed that FOXN3 was also downregulated in OS cells. Numerous functional experiments, including colony formation, Cell Counting Kit‑8, wound healing and Transwell invasion assays, were performed. The results of the present study revealed that FOXN3 suppressed the proliferation, migration and invasion of OS cells. SIRT6 has been reported to serve a key role in OS; chromatin‑immunoprecipitation (ChIP) and quantitative ChIP, as well as a luciferase reporter assay, demonstrated that SIRT6 was transcriptionally regulated by FOXN3. Furthermore, FOXN3 also regulated matrix metalloproteinase‑9 secretion via the regulation of SIRT6 expression. The findings of the present study indicated that FOXN3 serves as a tumor suppressor in OS and proposed FOXN3 as a prognostic predictor and a therapeutic target for patients with OS.

View Figures

View References

1	Bielack S, Carrle D and Casali PG; ESMO Guidelines Working Group, : Osteosarcoma: ESMO clinical recommendations for diagnosis, treatment and follow-up. Ann Oncol. 4 Suppl 20:S137–S139. 2009.
2	Damron TA, Ward WG and Stewart A: Osteosarcoma, chondrosarcoma, and Ewing's sarcoma: National cancer data base report. Clin Orthop Relat Res. 459:40–47. 2007. View Article : Google Scholar : PubMed/NCBI
3	Ottaviani G and Jaffe N: The epidemiology of osteosarcoma. Cancer Treat Res. 152:3–13. 2009. View Article : Google Scholar : PubMed/NCBI
4	Tan ML, Choong PF and Dass CR: Osteosarcoma: Conventional treatment vs. gene therapy. Cancer Biol Ther. 8:106–117. 2009. View Article : Google Scholar : PubMed/NCBI
5	Bacci G, Briccoli A, Rocca M, Ferrari S, Donati D, Longhi A, Bertoni F, Bacchini P, Giacomini S, Forni C, et al: Neoadjuvant chemotherapy for osteosarcoma of the extremities with metastases at presentation: Recent experience at the Rizzoli Institute in 57 patients treated with cisplatin, doxorubicin, and a high dose of methotrexate and ifosfamide. Ann Oncol. 14:1126–1134. 2003. View Article : Google Scholar : PubMed/NCBI
6	Rainusso N, Wang LL and Yustein JT: The adolescent and young adult with cancer: State of the art-bone tumors. Curr Oncol Rep. 15:296–307. 2013. View Article : Google Scholar : PubMed/NCBI
7	Bielack SS, Kempf-Bielack B, Delling G, Exner GU, Flege S, Helmke K, Kotz R, Salzer-Kuntschik M, Werner M, Winkelmann W, et al: Prognostic factors in high-grade osteosarcoma of the extremities or trunk: An analysis of 1,702 patients treated on neoadjuvant cooperative osteosarcoma study group protocols. J Clin Oncol. 20:776–790. 2002. View Article : Google Scholar : PubMed/NCBI
8	Scott KL and Plon SE: CHES1/FOXN3 interacts with Ski-interacting protein and acts as a transcriptional repressor. Gene. 359:119–126. 2005. View Article : Google Scholar : PubMed/NCBI
9	Sun J, Li H, Huo Q, Cui M, Ge C, Zhao F, Tian H, Chen T, Yao M and Li J: The transcription factor FOXN3 inhibits cell proliferation by downregulating E2F5 expression in hepatocellular carcinoma cells. Oncotarget. 7:43534–43545. 2016.PubMed/NCBI
10	Balciunaite G, Keller MP, Balciunaite E, Piali L, Zuklys S, Mathieu YD, Gill J, Boyd R, Sussman DJ and Holländer GA: Wnt glycoproteins regulate the expression of FoxN1, the gene defective in nude mice. Nat Immunol. 3:1102–1108. 2002. View Article : Google Scholar : PubMed/NCBI
11	Li C, Lusis AJ, Sparkes R, Tran SM and Gaynor R: Characterization and chromosomal mapping of the gene encoding the cellular DNA binding protein HTLF. Genomics. 13:658–664. 1992. View Article : Google Scholar : PubMed/NCBI
12	Chang JT, Wang HM, Chang KW, Chen WH, Wen MC, Hsu YM, Yung BY, Chen IH, Liao CT, Hsieh LL and Cheng AJ: Identification of differentially expressed genes in oral squamous cell carcinoma (OSCC): Overexpression of NPM, CDK1 and NDRG1 and underexpression of CHES1. Int J Cancer. 114:942–949. 2005. View Article : Google Scholar : PubMed/NCBI
13	Markowski J, Tyszkiewicz T, Jarzab M, Oczko-Wojciechowska M, Gierek T, Witkowska M, Paluch J, Kowalska M, Wygoda Z, Lange D and Jarzab B: Metal-proteinase ADAM12, kinesin 14 and checkpoint suppressor 1 as new molecular markers of laryngeal carcinoma. Eur Arch Otorhinolaryngol. 266:1501–1507. 2009. View Article : Google Scholar : PubMed/NCBI
14	Li S, Mo Z, Yang X, Price SM, Shen MM and Xiang M: Foxn4 controls the genesis of amacrine and horizontal cells by retinal progenitors. Neuron. 43:795–807. 2004. View Article : Google Scholar : PubMed/NCBI
15	Katoh M: Identification and characterization of human FOXN5 and rat Foxn5 genes in silico. Int J Oncol. 24:1339–1344. 2004.PubMed/NCBI
16	Wang X, He B, Gao Y and Li Y: FOXR2 contributes to cell proliferation and malignancy in human hepatocellular carcinoma. Tumour Biol. 37:10459–10467. 2016. View Article : Google Scholar : PubMed/NCBI
17	Huot G, Vernier M, Bourdeau V, Doucet L, Saint-Germain E, Gaumont-Leclerc MF, Moro A and Ferbeyre G: CHES1/FOXN3 regulates cell proliferation by repressing PIM2 and protein biosynthesis. Mol Biol Cell. 25:554–565. 2014. View Article : Google Scholar : PubMed/NCBI
18	Gertler AA and Cohen HY: SIRT6, a protein with many faces. Biogerontology. 14:629–639. 2013. View Article : Google Scholar : PubMed/NCBI
19	Kim HS, Xiao C, Wang RH, Lahusen T, Xu X, Vassilopoulos A, Vazquez-Ortiz G, Jeong WI, Park O, Ki SH, et al: Hepatic-specific disruption of SIRT6 in mice results in fatty liver formation due to enhanced glycolysis and triglyceride synthesis. Cell Metab. 12:224–236. 2010. View Article : Google Scholar : PubMed/NCBI
20	Lin H, Hao Y, Zhao Z and Tong Y: Sirtuin 6 contributes to migration and invasion of osteosarcoma cells via the ERK1/2/MMP9 pathway. FEBS Open Bio. 7:1291–1301. 2017. View Article : Google Scholar : PubMed/NCBI
21	Baumhoer D: Pathogenesis and genetics of osteosarcoma: Current concepts and developments. Der Pathologe. 39:139–145. 2018.(In German). View Article : Google Scholar : PubMed/NCBI
22	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 : PubMed/NCBI
23	Descamps FJ, Martens E and Opdenakker G: Analysis of gelatinases in complex biological fluids and tissue extracts. Lab Invest. 82:1607–1608. 2002. View Article : Google Scholar : PubMed/NCBI
24	Padala C, Tupurani MA, Puranam K, Gantala S, Shyamala N, Kondapalli MS, Gundapaneni KK, Mudigonda S, Galimudi RK, Kupsal K, et al: Synergistic effect of collagenase-1 (MMP1), stromelysin-1 (MMP3) and gelatinase-B (MMP9) gene polymorphisms in breast cancer. PLoS One. 12:e01844482017. View Article : Google Scholar : PubMed/NCBI
25	Cai X, Zhu H and Li Y: PKCzeta, MMP2 and MMP9 expression in lung adenocarcinoma and association with a metastatic phenotype. Mol Med Rep. 16:8301–8306. 2017. View Article : Google Scholar : PubMed/NCBI
26	Jia ZH, Jia Y, Guo FJ, Chen J, Zhang XW and Cui MH: Phosphorylation of STAT3 at Tyr705 regulates MMP-9 production in epithelial ovarian cancer. PLoS One. 12:e01836222017. View Article : Google Scholar : PubMed/NCBI
27	Bai L, Lin G, Sun L, Liu Y, Huang X, Cao C, Guo Y and Xie C: Upregulation of SIRT6 predicts poor prognosis and promotes metastasis of non-small cell lung cancer via the ERK1/2/MMP9 pathway. Oncotarget. 7:40377–40386. 2016. View Article : Google Scholar : PubMed/NCBI
28	Schuff M, Rossner A, Wacker SA, Donow C, Gessert S and Knochel W: FoxN3 is required for craniofacial and eye development of Xenopus laevis. Dev Dyn. 236:226–239. 2007. View Article : Google Scholar : PubMed/NCBI
29	Samaan G, Yugo D, Rajagopalan S, Wall J, Donnell R, Goldowitz D, Gopalakrishnan R and Venkatachalam S: Foxn3 is essential for craniofacial development in mice and a putative candidate involved in human congenital craniofacial defects. Biochem Biophys Res Commun. 400:60–65. 2010. View Article : Google Scholar : PubMed/NCBI
30	Nagel S, Meyer C, Kaufmann M, Drexler HG and MacLeod RA: Deregulated FOX genes in Hodgkin lymphoma. Genes Chromosomes Cancer. 53:917–933. 2014. View Article : Google Scholar : PubMed/NCBI
31	Cai Y, Mohseny AB, Karperien M, Hogendoorn PC, Zhou G and Cleton-Jansen AM: Inactive Wnt/beta-catenin pathway in conventional high-grade osteosarcoma. J Pathol. 220:24–33. 2010. View Article : Google Scholar : PubMed/NCBI
32	Hendrix ND, Wu R, Kuick R, Schwartz DR, Fearon ER and Cho KR: Fibroblast growth factor 9 has oncogenic activity and is a downstream target of Wnt signaling in ovarian endometrioid adenocarcinomas. Cancer Res. 66:1354–1362. 2006. View Article : Google Scholar : PubMed/NCBI
33	Du Z, Li F, Wang L, Huang H and Xu S: Regulatory effects of microRNA184 on osteosarcoma via the Wnt/β-catenin signaling pathway. Mol Med Rep. 18:1917–1924. 2018.PubMed/NCBI
34	Zhao X, Sun S, Xu J, Luo Y, Xin Y and Wang Y: MicroRNA-152 inhibits cell proliferation of osteosarcoma by directly targeting Wnt/β-catenin signaling pathway in a DKK1-dependent manner. Oncol Rep. 40:767–774. 2018.PubMed/NCBI
35	Yu M, Guo D, Cao Z, Xiao L and Wang G: Inhibitory effect of microRNA-107 on osteosarcoma malignancy through regulation of Wnt/β-catenin signaling in vitro. Cancer Invest. 36:175–184. 2018. View Article : Google Scholar : PubMed/NCBI