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Open Access 01-12-2023 | Glioma | Research

Glioma angiogenesis is boosted by ELK3 activating the HIF-1\(\alpha\)/VEGF-A signaling axis

Authors: Mou Yueyang, Hu Yaqin, Xue Guolian, Zhao Wenjian, Jiao Yang, Li Chen, Cao Haiyan, Chao Min, Deng Jianping, Dai Penggao, Zhu Hongli, Wang Liang

Published in: BMC Cancer | Issue 1/2023

Abstract

Background

Clinical studies have shown that first-line use of anti-angiogenetic therapy can prolong progression-free survival but little progress has been made in extending the overall survival of the patients. We explored the role of ELK3 in glioma angiogenesis to improve and design more efficacious therapies.

Methods

A tissue microarray and immunohistochemistry analysis were used to determine the expression of ELK3 protein in 400 glioma patients. Cell proliferation, metastasis, cell cycle, and apoptosis were monitored in U87 and U251 cells using CCK-8, EdU, transwell assays, and flow cytometry. A tube-formation assay, a rat aorta ring sprouting assay, and a matrigel plug assay were performed to examine the antiangiogenic activity of ELK3. An ELISA, Western blot, and correlation analysis of the CGGA dataset were used to detect the association between ELK3 and VEGF-A or ELK3 and HIF-1\(\alpha\). Besides, orthotopic transplantation in nude mice and histopathological and immunological analysis of in vitro tumors were used to explore the effect of ELK3 on tumor progression and median survival.

Results

ELK3 was upregulated in glioma tissues and associated with a poor prognosis. In vitro, ELK3 promoted cell proliferation and cell cycle progression, induced metastasis, and suppressed apoptosis. Then, silencing ELK3 inhibited VEGF-A expression and secretion by facilitating HIF-1\(\alpha\) degradation via ubiquitination. Finally, knockdown ELK3 inhibited tumor progression and angiogenesis in vitro and in vivo, as well as prolonged nude mice’s median survival.

Conclusions

Our findings first evidenced that ELK3 is crucial for glioma because it promotes angiogenesis by activating the HIF-1\(\alpha\)/VEGF-A signaling axis. Therefore, we suggest that ELK3 is a prognostic marker with a great potential for glioma angiogenesis and ELK3-targeted therapeutic strategies might hold promise in improving the efficacy of anti-angiogenic therapies.

Ostrom QT, Bauchet L, Davis FG, Deltour I, Fisher JL, Langer CE, et al. The epidemiology of glioma in adults: a “state of the science’’ review. Neuro-Oncol. 2014;16(7):896–913. https://doi.org/10.1093/neuonc/nou087.CrossRefPubMedPubMedCentral

Kim BS, Seol HJ, Nam DH, Park CK, Kim IH, Kim TM, et al. Concurrent Chemoradiotherapy with Temozolomide Followed by Adjuvant Temozolomide for Newly Diagnosed Glioblastoma Patients: A Retrospective Multicenter Observation Study in Korea. Cancer Res Treat. 2017;49(1):193–203. https://doi.org/10.4143/crt.2015.473.CrossRefPubMed

Lang HL, Hu GW, Zhang B, Kuang W, Chen Y, Wu L, et al. Glioma cells enhance angiogenesis and inhibit endothelial cell apoptosis through the release of exosomes that contain long non-coding RNA CCAT2. Oncol Rep. 2017;38(2):785–98. https://doi.org/10.3892/or.2017.5742.CrossRefPubMedPubMedCentral

Gilbert MR, Dignam JJ, Armstrong TS, Wefel JS, Blumenthal DT, Vogelbaum MA, et al. A randomized trial of bevacizumab for newly diagnosed glioblastoma. New England J Med. 2014;370(8):699–708. https://doi.org/10.1056/nejmoa1308573.CrossRef

Rosińska S, Gavard J. Tumor Vessels Fuel the Fire in Glioblastoma. Int J Mol Sci. 2021;22(12):6514. https://doi.org/10.3390/ijms22126514.CrossRefPubMedPubMedCentral

Wang X, Li X, Dai X, Zhang X, Zhang J, Xu T, et al. Bioprinting of glioma stem cells improves their endotheliogenic potential. Colloids Surf B Biointerfaces. 2018;171:629–37. https://doi.org/10.1016/j.colsurfb.2018.08.006.CrossRefPubMed

Heng D, Mackenzie M, Vaishampayan U, Bjarnason G, Knox J, Tan M, et al. Primary anti-vascular endothelial growth factor (VEGF)-refractory metastatic renal cell carcinoma: clinical characteristics, risk factors, and subsequent therapy. Ann Oncol Off J Eur Soc Med Oncol. 2012;23(6):1549–55. https://doi.org/10.1093/annonc/mdr533.CrossRef

Hirota K, Semenza GL. Regulation of angiogenesis by hypoxia-inducible factor 1. Crit Rev Oncol Hematol. 2006;59(1):15–26. https://doi.org/10.1016/j.critrevonc.2005.12.003.CrossRefPubMed

Koukourakis MI, Giatromanolaki A, Sivridis E, Simopoulos C, Turley H, Talks K, et al. Hypoxia-inducible factor (HIF1A and HIF2A), angiogenesis, and chemoradiotherapy outcome of squamous cell head-and-neck cancer. Int J Radiat Oncol Biol Phys. 2002;53(5):1192–202. https://doi.org/10.1016/S0360-3016(02)02848-1.CrossRefPubMed

10.

Giovane A, Pintzas A, Maira S, Sobieszczuk P, Wasylyk B. Net, a new ets transcription factor that is activated by Ras. Genes Dev. 1994;8(13):1502–13. https://doi.org/10.1101/gad.8.13.1502.CrossRefPubMed

11.

Ayadi A, Suelves M, Dollé P, Wasylyk B. Net, an Ets ternary complex transcription factor, is expressed in sites of vasculogenesis, angiogenesis, and chondrogenesis during mouse development. Mech Dev. 2001;102(1):205–8. https://doi.org/10.1016/S0925-4773(01)00289-1.CrossRefPubMed

12.

Weinl C, Wasylyk C, Garcia Garrido M, Sothilingam V, Beck SC, Riehle H, et al. Elk3 deficiency causes transient impairment in post-natal retinal vascular development and formation of tortuous arteries in adult murine retinae. PLoS ONE. 2014;9(9):e107048. https://doi.org/10.1371/journal.pone.0107048.CrossRefPubMedPubMedCentral

13.

Park JI, Kim KS, Kong SY, Park KS. Novel function of E26 transformation-specific domain-containing protein ELK3 in lymphatic endothelial cells. Oncol Lett. 2018;15(1):55–60. https://doi.org/10.3892/ol.2017.7308.CrossRefPubMed

14.

Kim KS, Park JI, Oh N, Cho HJ, Park JH, Park KS. ELK3 expressed in lymphatic endothelial cells promotes breast cancer progression and metastasis through exosomal miRNAs. Sci Rep. 2019;9(1):8418. https://doi.org/10.1038/s41598-019-44828-6.CrossRefPubMedPubMedCentral

15.

Yang H, Schramek D, Adam RC, Keyes BE, Wang P, Zheng D, et al. ETS family transcriptional regulators drive chromatin dynamics and malignancy in squamous cell carcinomas. eLife. 2015;4:e10870. https://doi.org/10.7554/elife.10870.CrossRefPubMedPubMedCentral

16.

Acerbi I, Cassereau L, Dean I, Shi Q, Au A, Park C, et al. Human breast cancer invasion and aggression correlates with ECM stiffening and immune cell infiltration. Integr Biol Quant Biosci Nano Macro. 2015;7(10):1120–34. https://doi.org/10.1039/c5ib00040h.CrossRef

17.

Han B, Zhang H, Tian R, Liu H, Wang Z, Wang Z, et al. Exosomal EPHA2 derived from highly metastatic breast cancer cells promotes angiogenesis by activating the AMPK signaling pathway through Ephrin A1-EPHA2 forward signaling. Theranostics. 2022;12:4127–46. https://doi.org/10.7150/thno.72404.CrossRefPubMedPubMedCentral

18.

Zhi T, Jiang K, Zhang C, Xu X, Wu W, Nie E, et al. MicroRNA-1301 inhibits proliferation of human glioma cells by directly targeting N-Ras. Am J Cancer Res. 2017;7(4):982–98.PubMedPubMedCentral

19.

Fu S, Hou MM, Naing A, Janku F, Hess K, Zinner R, et al. Phase I study of pazopanib and vorinostat: a therapeutic approach for inhibiting mutant p53-mediated angiogenesis and facilitating mutant p53 degradation. Ann Oncol. 2015;26(5):1012–8. https://doi.org/10.1093/annonc/mdv066.CrossRefPubMedPubMedCentral

20.

Lee JH, Hur W, Hong SW, Kim JH, Kim SM, Lee EB, et al. ELK3 promotes the migration and invasion of liver cancer stem cells by targeting HIF-1\(\alpha\). Oncol Rep. 2017;37(2):813–22. https://doi.org/10.3892/or.2016.5293.

21.

Semenza GL. Targeting HIF-1 for cancer therapy. Nat Rev Cancer. 2003;3(10):721–32. https://doi.org/10.1038/nrc1187.CrossRefPubMed

22.

Liu Z, Ren Z, Zhang C, Qian R, Wang H, Wang J, et al. ELK3: A New Molecular Marker for the Diagnosis and Prognosis of Glioma. Front Oncol. 2021;11. https://doi.org/10.3389/fonc.2021.608748.

23.

Zhao Q, Ren Y, Xie H, Yu L, Lu J, Jiang W, et al. ELK3 Mediated by ZEB1 Facilitates the Growth and Metastasis of Pancreatic Carcinoma by Activating the Wnt/\(\beta\)-Catenin Pathway. Front Dev Biol. 2021;9. https://doi.org/10.3389/fcell.2021.700192.

24.

Meng L, Xing Z, Guo Z, Liu Z. LINC01106 post-transcriptionally regulates ELK3 and HOXD8 to promote bladder cancer progression. Cell Death Dis. 2020;11(12):1063. https://doi.org/10.1038/s41419-020-03236-9.CrossRefPubMedPubMedCentral

25.

Heo SH, Lee JY, Yang KM, Park KS. ELK3 Expression Correlates With Cell Migration, Invasion, and Membrane Type 1-Matrix Metalloproteinase Expression in MDA-MB-231 Breast Cancer Cells. Gene Expr. 2015;16(4):197–203. https://doi.org/10.3727/105221615x14399878166276.CrossRefPubMedPubMedCentral

26.

Plate K, Breier G, Weich H, Risau W. Vascular endothelial growth factor is a potential tumour angiogenesis factor in human gliomas in vivo. Nature. 1992;359(6398):845–8. https://doi.org/10.1038/359845a0.CrossRefPubMed

27.

Wick W, Platten M, Wick A, Berberich A, Radbruch A, Bendszus M, et al. Current status and future directions of anti-angiogenic therapy for gliomas. Neuro-Oncol. 2015;18. https://doi.org/10.1093/neuonc/nov180.

28.

Maira S, Wurtz J, Wasylyk B. Net (ERP/SAP2) one of the Ras-inducible TCFs, has a novel inhibitory domain with resemblance to the helix-loop-helix motif. EMBO J. 1996;15(21):5849–65. https://doi.org/10.1002/j.1460-2075.1996.tb00972.x.CrossRefPubMedPubMedCentral

29.

Chinot OL, Wick W, Mason W, Henriksson R, Saran F, Nishikawa R, et al. Bevacizumab plus Radiotherapy-Temozolomide for Newly Diagnosed Glioblastoma. N Engl J Med. 2014;370(8):709–22. https://doi.org/10.1056/NEJMoa1308345.CrossRefPubMed

30.

Sandmann T, Bourgon R, Garcia J, Li C, Cloughesy T, Chinot OL, et al. Patients With Proneural Glioblastoma May Derive Overall Survival Benefit From the Addition of Bevacizumab to First-Line Radiotherapy and Temozolomide: Retrospective Analysis of the AVAglio Trial. J Clin Oncol Off J Am Soc Clin Oncol. 2015;33(25):2735–44. https://doi.org/10.1200/jco.2015.61.5005.CrossRef

31.

Lai A, Tran A, Nghiemphu PL, Pope WB, Solis OE, Selch M, et al. Phase II study of bevacizumab plus temozolomide during and after radiation therapy for patients with newly diagnosed glioblastoma multiforme. J Clin Oncol Off J Am Soc Clin Oncol. 2011;29(2):142–8. https://doi.org/10.1200/jco.2010.30.2729.CrossRef

32.

Goldbrunner R, Wagner S, Roosen K, Tonn J. Models for assessment of angiogenesis in gliomas. J Neuro-Oncol. 2000;50(1–2):53–62. https://doi.org/10.1023/a:1006462504447.CrossRef

33.

Baker GJ, Yadav VN, Motsch S, Koschmann C, Calinescu AA, Mineharu Y, et al. Mechanisms of Glioma Formation: Iterative Perivascular Glioma Growth and Invasion Leads to Tumor Progression, VEGF-Independent Vascularization, and Resistance to Antiangiogenic Therapy. Neoplasia. 2014;16(7):543–61. https://doi.org/10.1016/j.neo.2014.06.003.CrossRefPubMedPubMedCentral

34.

Ribatti D. Novel angiogenesis inhibitors: addressing the issue of redundancy in the angiogenic signaling pathway. Cancer Treat Rev. 2011;37(5):344–52. https://doi.org/10.1016/j.ctrv.2011.02.002.CrossRefPubMed

35.

Osterberg N, Ferrara N, Vacher J, Gaedicke S, Niedermann G, Weyerbrock A, et al. Decrease of VEGF-A in myeloid cells attenuates glioma progression and prolongs survival in an experimental glioma model. Neuro-Oncol. 2016;18(7):939–49. https://doi.org/10.1093/neuonc/now005.CrossRefPubMedPubMedCentral

36.

Ma X, Li Z, Li T, Zhu L, Li Z, Tian N. Long non-coding RNA HOTAIR enhances angiogenesis by induction of VEGFA expression in glioma cells and transmission to endothelial cells via glioma cell derived-extracellular vesicles. Am J Transl Res. 2017;9(11):5012–21.PubMedPubMedCentral

37.

Masoud GN, Li W. HIF-1\(\alpha\) pathway: role, regulation and intervention for cancer therapy. Acta Pharm Sin B. 2015;5(5):378–89. https://doi.org/10.1016/j.apsb.2015.05.007.

38.

Ravi R, Mookerjee B, Bhujwalla Z, Sutter C, Artemov D, Zeng Q, et al. Regulation of tumor angiogenesis by p53-induced degradation of hypoxia-inducible factor 1alpha. Genes Dev. 2000;14(1):34–44.CrossRefPubMedPubMedCentral

39.

Palazon A, Tyrakis PA, Macias D, Veliça P, Rundqvist H, Fitzpatrick S, et al. An HIF-1\(\alpha\)/VEGF-A Axis in Cytotoxic T Cells Regulates Tumor Progression. Cancer Cell. 2017;32(5):669-683.e5. https://doi.org/10.1016/j.ccell.2017.10.003.

40.

Liu Z, Tian Y, Wu H, Ouyang S, Kuang W. LncRNA H19 promotes glioma angiogenesis through miR-138/HIF-1\(\alpha\)/VEGF axis. Neoplasma. 2019;67(1):111–8.

Title: Glioma angiogenesis is boosted by ELK3 activating the HIF-1/VEGF-A signaling axis
Authors: Mou Yueyang
Hu Yaqin
Xue Guolian
Zhao Wenjian
Jiao Yang
Li Chen
Cao Haiyan
Chao Min
Deng Jianping
Dai Penggao
Zhu Hongli
Wang Liang
Publication date: 01-12-2023
Publisher: BioMed Central
Keywords: Glioma
Glioma
Glioma
Published in: BMC Cancer / Issue 1/2023
Electronic ISSN: 1471-2407
DOI: https://doi.org/10.1186/s12885-023-11069-w

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