Role of MYC-miR-29-B7-H3 in Medulloblastoma Growth and Angiogenesis
Abstract
:1. Introduction
2. Materials and Methods
2.1. Ethics Statement
2.2. In Silico Analysis
2.3. Cell Culture and Transfection
2.4. Plasmids, Antibodies, and Chemical Inhibitors
2.5. ELISA, Human Angiogenesis Array, Immunoblotting and Gelatin Zymography
2.6. F-actin Staining and Immunostaining
2.7. Real-Time PCR and RNA-Seq
2.8. IPAD Analysis
2.9. Chromatin Immunoprecipitation (ChIP) and DNA Sequencing
2.10. Fluorescence-Activated Cell Sorting (FACS) Analysis
2.11. In-Vitro Angiogenesis Assay
2.12. Chick Chorioallantoic Membrane Assay (CAM)
2.13. Statistical Analysis
3. Results
3.1. High Expression of B7-H3 Is Associated with Poor Survival in MB Patients
3.2. B7-H3 Can Be Regulated via Myc-miR-29 Axis in MB Cells
3.3. MYC May Regulate B7-H3 Transcription
3.4. B7-H3 Promotes Angiogenesis in MB Cells
3.5. Role of sB7-H3 in Angiogenesis
3.6. miR-29 Exhibits Global Anti-Tumor Functions and Promotes STAT1 Activation
3.7. STAT1 Activity Downregulates MYC and B7-H3 Expression in MB Cells
3.8. miR-29 in Combination with MYC Inhibition Induces Apoptosis
4. Discussion
5. Conclusion
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
- Cassia, G.S.E.; Alves, C.A.P.F.; Taranath, A.; López, N.S.; Oztekin, O.; Gonçalves, F.G.; Patay, Z. Childhood Medulloblastoma Revisited. Top Magn. Reson. Imaging. 2018, 27, 479–502. [Google Scholar] [CrossRef] [PubMed]
- Kuzan-Fischer, C.M.; Juraschka, K.; Taylor, M.D. Medulloblastoma in the Molecular Era. J. Korean Neurosurg. Soc. 2018, 61, 292–301. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Schwalbe, E.C.; Lindsey, J.C.; Nakjang, S.; Crosier, S.; Smith, A.J.; Hicks, D.; Rafiee, G.; Hill, R.M.; Iliasova, A.; Stone, T.; et al. Novel molecular subgroups for clinical classification and outcome prediction in childhood medulloblastoma: A cohort study. Lancet. Oncol. 2017, 18, 958–971. [Google Scholar] [CrossRef]
- Roussel, M.F.; Robinson, G.W. Role of MYC in Medulloblastoma. Cold Spring Harb. Perspect Med. 2013, 3. [Google Scholar] [CrossRef] [PubMed]
- Pfister, S.; Remke, M.; Benner, A.; Mendrzyk, F.; Toedt, G.; Felsberg, J.; Wittmann, A.; Devens, F.; Gerber, N.U.; Joos, S.; et al. Outcome prediction in pediatric medulloblastoma based on DNA copy-number aberrations of chromosomes 6q and 17q and the MYC and MYCN loci. J. Clin. Oncol. 2009, 27, 1627–1636. [Google Scholar] [CrossRef] [PubMed]
- Staal, J.A.; Pei, Y.; Rood, B.R. A Proteogenomic Approach to Understanding MYC Function in Metastatic Medulloblastoma Tumors. Int. J. Mol. Sci. 2016, 17, 1744. [Google Scholar] [CrossRef] [PubMed]
- Mazzoccoli, L.; Robaina, M.C.; Apa, A.G.; Bonamino, M.; Pinto, L.W.; Queiroga, E.; Bacchi, C.E.; Klumb, C.E. MiR-29 silencing modulates the expression of target genes related to proliferation, apoptosis and methylation in Burkitt lymphoma cells. J. Cancer Res. Clin. Oncol. 2018, 144, 483–497. [Google Scholar] [CrossRef] [PubMed]
- Hashimoto, M.; Suizu, F.; Tokuyama, W.; Noguchi, H.; Hirata, N.; Matsuda-Lennikov, M.; Edamura, T.; Masuzawa, M.; Gotoh, N.; Tanaka, S.; et al. Protooncogene TCL1b functions as an Akt kinase co-activator that exhibits oncogenic potency in vivo. Oncogenesis 2013, 2, e70. [Google Scholar] [CrossRef] [PubMed]
- Zhu, K.; Liu, L.; Zhang, J.; Wang, Y.; Liang, H.; Fan, G.; Jiang, Z.; Zhang, C.Y.; Chen, X.; Zhou, G. MiR-29b suppresses the proliferation and migration of osteosarcoma cells by targeting CDK6. Protein Cell 2016, 7, 434–444. [Google Scholar] [CrossRef] [Green Version]
- Zhang, X.; Zhao, X.; Fiskus, W.; Lin, J.; Lwin, T.; Rao, R.; Zhang, Y.; Chan, J.C.; Fu, K.; Marquez, V.E.; et al. Coordinated silencing of MYC-mediated miR-29 by HDAC3 and EZH2 as a therapeutic target of histone modification in aggressive B-Cell lymphomas. Cancer Cell 2012, 22, 506–523. [Google Scholar] [CrossRef]
- Garzon, R.; Liu, S.; Fabbri, M.; Liu, Z.; Heaphy, C.E.; Callegari, E.; Schwind, S.; Pang, J.; Yu, J.; Muthusamy, N.; et al. MicroRNA-29b induces global DNA hypomethylation and tumor suppressor gene reexpression in acute myeloid leukemia by targeting directly DNMT3A and 3B and indirectly DNMT1. Blood 2009, 113, 6411–6418. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Shin, J.; Shim, H.G.; Hwang, T.; Kim, H.; Kang, S.H.; Dho, Y.S.; Park, S.H.; Kim, S.J.; Park, C.K. Restoration of miR-29b exerts anti-cancer effects on glioblastoma. Cancer Cell Int. 2017, 17, 104. [Google Scholar] [CrossRef] [PubMed]
- Gregorio, A.; Corrias, M.V.; Castriconi, R.; Dondero, A.; Mosconi, M.; Gambini, C.; Moretta, A.; Moretta, L.; Bottino, C. Small round blue cell tumours: Diagnostic and prognostic usefulness of the expression of B7-H3 surface molecule. Histopathology 2008, 53, 73–80. [Google Scholar] [CrossRef] [PubMed]
- Ma, J.; Ma, P.; Zhao, C.; Xue, X.; Han, H.; Liu, C.; Tao, H.; Xiu, W.; Cai, J.; Zhang, M. B7-H3 as a promising target for cytotoxicity T cell in human cancer therapy. Oncotarget 2016, 7, 29480–29491. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Chen, Y.W.; Tekle, C.; Fodstad, O. The immunoregulatory protein human B7H3 is a tumor-associated antigen that regulates tumor cell migration and invasion. Curr. Cancer Drug Targets 2008, 8, 404–413. [Google Scholar] [CrossRef] [PubMed]
- Vigdorovich, V.; Ramagopal, U.A.; Lazar-Molnar, E.; Sylvestre, E.; Lee, J.S.; Hofmeyer, K.A.; Zang, X.; Nathenson, S.G.; Almo, S.C. Structure and T cell inhibition properties of B7 family member, B7-H3. Structure 2013, 21, 707–717. [Google Scholar] [CrossRef] [PubMed]
- Wang, S.; Mou, J.; Cui, L.; Wang, X.; Zhang, Z. Astragaloside IV inhibits cell proliferation of colorectal cancer cell lines through down-regulation of B7-H3. Biomed. Pharmacother 2018, 102, 1037–1044. [Google Scholar] [CrossRef] [PubMed]
- Li, Y.; Zhang, J.; Han, S.; Qian, Q.; Chen, Q.; Liu, L.; Zhang, Y. B7-H3 promotes the proliferation, migration and invasiveness of cervical cancer cells and is an indicator of poor prognosis. Oncol. Rep. 2017, 38, 1043–1050. [Google Scholar] [CrossRef]
- Majzner, R.G.; Theruvath, J.L.; Nellan, A.; Heitzeneder, S.; Cui, Y.; Mount, C.W.; Rietberg, S.P.; Linde, M.H.; Xu, P.; Rota, C.; et al. CAR T Cells Targeting B7-H3, a Pan-Cancer Antigen, Demonstrate Potent Preclinical Activity Against Pediatric Solid Tumors and Brain Tumors. Clin. Cancer Res. 2019, 25, 2560–2574. [Google Scholar] [CrossRef]
- Asuthkar, S.; Elustondo, P.A.; Demirkhanyan, L.; Sun, X.; Baskaran, P.; Velpula, K.K.; Thyagarajan, B.; Pavlov, E.V.; Zakharian, E. The TRPM8 protein is a testosterone receptor: I. Biochemical evidence for direct TRPM8-testosterone interactions. J. Biol. Chem. 2015, 290, 2659–2669. [Google Scholar] [CrossRef]
- Asuthkar, S.; Velpula, K.K.; Nalla, A.K.; Gogineni, V.R.; Gondi, C.S.; Rao, J.S. Irradiation-induced angiogenesis is associated with an MMP-9-miR-494-syndecan-1 regulatory loop in medulloblastoma cells. Oncogene 2014, 33, 1922–1933. [Google Scholar] [CrossRef] [PubMed]
- Asuthkar, S.; Gondi, C.S.; Nalla, A.K.; Velpula, K.K.; Gorantla, B.; Rao, J.S. Urokinase-type plasminogen activator receptor (uPAR)-mediated regulation of WNT/beta-catenin signaling is enhanced in irradiated medulloblastoma cells. J. Biol. Chem. 2012, 287, 20576–20589. [Google Scholar] [CrossRef] [PubMed]
- Gorantla, B.; Asuthkar, S.; Rao, J.S.; Patel, J.; Gondi, C.S. Suppression of the uPAR-uPA system retards angiogenesis, invasion, and in vivo tumor development in pancreatic cancer cells. Mol. Cancer Res. 2011, 9, 377–389. [Google Scholar] [CrossRef] [PubMed]
- Boyineni, J.; Tanpure, S.; Gnanamony, M.; Antony, R.; Fernández, K.S.; Lin, J.; Pinson, D.; Gondi, C.S. SPARC overexpression combined with radiation retards angiogenesis by suppressing VEGF-A via miR410 in human neuroblastoma cells. Int. J. Oncol. 2016, 49, 1394–1406. [Google Scholar] [CrossRef] [PubMed]
- Castellanos, J.R.; Purvis, I.J.; Labak, C.M.; Guda, M.R.; Tsung, A.J.; Velpula, K.K.; Asuthkar, S. B7-H3 role in the immune landscape of cancer. Am. J. Clin. Exp. Immunol. 2017, 6, 66–75. [Google Scholar] [PubMed]
- Delmore, J.E.; Issa, G.C.; Lemieux, M.E.; Rahl, P.B.; Shi, J.; Jacobs, H.M.; Kastritis, E.; Gilpatrick, T.; Paranal, R.M.; Qi, J.; et al. BET bromodomain inhibition as a therapeutic strategy to target c-Myc. Cell 2011, 146, 904–917. [Google Scholar] [CrossRef]
- McKeown, M.R.; Bradner, J.E. Therapeutic strategies to inhibit MYC. Cold Spring Harb. Perspect. Med. 2014, 4. [Google Scholar] [CrossRef] [PubMed]
- Xu, H.; Cheung, I.Y.; Guo, H.F.; Cheung, N.K. MicroRNA miR-29 modulates expression of immunoinhibitory molecule B7-H3: potential implications for immune based therapy of human solid tumors. Cancer Res. 2009, 69, 6275–6281. [Google Scholar] [CrossRef]
- Nygren, M.K.; Tekle, C.; Ingebrigtsen, V.A.; Mäkelä, R.; Krohn, M.; Aure, M.R.; Nunes-Xavier, C.E.; Perälä, M.; Tramm, T.; Alsner, J.; et al. Identifying microRNAs regulating B7-H3 in breast cancer: The clinical impact of microRNA-29c. Br. J. Cancer 2014, 110, 2072–2080. [Google Scholar] [CrossRef]
- Chiew, G.G.Y.; Fu, A.; Low, K.P.; Luo, K.Q. Physical supports from liver cancer cells are essential for differentiation and remodeling of endothelial cells in a HepG2-HUVEC co-culture model. Sci Rep. 2015, 5, 10801. [Google Scholar] [CrossRef]
- Chen, W.; Liu, P.; Wang, Y.; Nie, W.; Li, Z.; Xu, W.; Li, F.; Zhou, Z.; Zhao, M.; Liu, H. Characterization of a soluble B7-H3 (sB7-H3) spliced from the intron and analysis of sB7-H3 in the sera of patients with hepatocellular carcinoma. PLoS ONE 2013, 8, e76965. [Google Scholar] [CrossRef] [PubMed]
- Wang, L.; Kang, F.B.; Zhang, G.C.; Wang, J.; Xie, M.F.; Zhang, Y.Z. Clinical significance of serum soluble B7-H3 in patients with osteosarcoma. Cancer Cell Int. 2018, 18, 115. [Google Scholar] [CrossRef] [PubMed]
- Xie, C.; Liu, D.; Chen, Q.; Yang, C.; Wang, B.; Wu, H. Soluble B7-H3 promotes the invasion and metastasis of pancreatic carcinoma cells through the TLR4/NF-kappaB pathway. Sci. Rep. 2016, 6, 27528. [Google Scholar] [CrossRef] [PubMed]
- Li, Y.; Guo, G.; Song, J.; Cai, Z.; Yang, J.; Chen, Z.; Wang, Y.; Huang, Y.; Gao, Q. B7-H3 Promotes the Migration and Invasion of Human Bladder Cancer Cells via the PI3K/Akt/STAT3 Signaling Pathway. J. Cancer 2017, 8, 816–824. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Wang, L.; Zhang, Q.; Chen, W.; Shan, B.; Ding, Y.; Zhang, G.; Cao, N.; Liu, L.; Zhang, Y. B7-H3 is overexpressed in patients suffering osteosarcoma and associated with tumor aggressiveness and metastasis. PLoS ONE 2013, 8, e70689. [Google Scholar] [CrossRef] [PubMed]
- Chen, Y.; Fang, L.; Zhang, J.; Li, G.; Ma, M.; Li, C.; Lyu, J.; Meng, Q. Blockage of Glyoxalase I Inhibits Colorectal Tumorigenesis and Tumor Growth via Upregulation of STAT1, p53, and Bax and Downregulation of c-Myc and Bcl-2. Int. J. Mol. Sci. 2017, 18, 570. [Google Scholar] [CrossRef]
- Zheng, J.; van de Veerdonk, F.L.; Crossland, K.L.; Smeekens, S.P.; Chan, C.M.; Al Shehri, T.; Abinun, M.; Gennery, A.R.; Mann, J.; Lendrem, D.W.; et al. Gain-of-function STAT1 mutations impair STAT3 activity in patients with chronic mucocutaneous candidiasis (CMC). Eur. J. Immunol. 2015, 45, 2834–2846. [Google Scholar] [CrossRef] [PubMed]
- Levy, D.E.; Darnell, J.E., Jr. Signalling: Stats: Transcriptional control and biological impact. Nat. Rev. Mol. Cell Biol. 2002, 3, 651–662. [Google Scholar] [CrossRef]
- Blum, W.; Schwind, S.; Tarighat, S.S.; Geyer, S.; Eisfeld, A.K.; Whitman, S.; Walker, A.; Klisovic, R.; Byrd, J.C.; Santhanam, R.; et al. Clinical and pharmacodynamic activity of bortezomib and decitabine in acute myeloid leukemia. Blood 2012, 119, 6025–6031. [Google Scholar] [CrossRef]
- Mihailovich, M.; Bremang, M.; Spadotto, V.; Musiani, D.; Vitale, E.; Varano, G.; Zambelli, F.; Mancuso, F.M.; Cairns, D.A.; Pavesi, G.; et al. miR-17-92 fine-tunes MYC expression and function to ensure optimal B cell lymphoma growth. Nat. Commun. 2015, 6, 8725. [Google Scholar] [CrossRef]
- Jiang, H.; Zhang, G.; Wu, J.H.; Jiang, C.P. Diverse roles of miR-29 in cancer (review). Oncol. Rep. 2014, 31, 1509–1516. [Google Scholar] [CrossRef] [PubMed]
- Xu, L.; Ding, X.; Tan, H.; Qian, J. Correlation between B7-H3 expression and matrix metalloproteinases 2 expression in pancreatic cancer. Cancer Cell Int. 2013, 13, 81. [Google Scholar] [CrossRef] [PubMed]
- Mesri, M.; Birse, C.; Heidbrink, J.; McKinnon, K.; Brand, E.; Bermingham, C.L.; Feild, B.; FitzHugh, W.; He, T.; Ruben, S.; et al. Identification and characterization of angiogenesis targets through proteomic profiling of endothelial cells in human cancer tissues. PLoS ONE 2013, 8, e78885. [Google Scholar] [CrossRef] [PubMed]
- Wang, F.; Ke, Z.F.; Wang, R.; Wang, Y.F.; Huang, L.L.; Wang, L.T. Astrocyte elevated gene-1 (AEG-1) promotes osteosarcoma cell invasion through the JNK/c-Jun/MMP-2 pathway. Biochem. Biophys. Res. Commun. 2014, 452, 933–939. [Google Scholar] [CrossRef] [PubMed]
- Chen, X.; Quinn, E.M.; Ni, H.; Wang, J.; Blankson, S.; Redmond, H.P.; Wang, J.H.; Feng, X. B7-H3 participates in the development of experimental pneumococcal meningitis by augmentation of the inflammatory response via a TLR2-dependent mechanism. J. Immunol. 2012, 189, 347–355. [Google Scholar] [CrossRef] [PubMed]
- Cantley, L.C.; Neel, B.G. New insights into tumor suppression: PTEN suppresses tumor formation by restraining the phosphoinositide 3-kinase/AKT pathway. Proc. Natl. Acad. Sci. USA 1999, 96, 4240–4245. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Palmbos, P.L.; Wang, L.; Yang, H.; Wang, Y.; Leflein, J.; Ahmet, M.L.; Wilkinson, J.E.; Kumar-Sinha, C.; Ney, G.M.; Tomlins, S.A.; et al. ATDC/TRIM29 Drives Invasive Bladder Cancer Formation through miRNA-Mediated and Epigenetic Mechanisms. Cancer Res. 2015, 75, 5155–5166. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Yang, N.; Gilman, P.; Mirzayans, R.; Sun, X.; Touret, N.; Weinfeld, M.; Goping, S. Characterization of the apoptotic response induced by the cyanine dye D112: A potentially selective anti-cancer compound. PLoS ONE 2015, 10, e0125381. [Google Scholar] [CrossRef] [PubMed]
- Yu, S.W.; Andrabi, S.A.; Wang, H.; Kim, N.S.; Poirier, G.G.; Dawson, T.M.; Dawson, V.L. Apoptosis-inducing factor mediates poly(ADP-ribose) (PAR) polymer-induced cell death. Proc. Natl. Acad. Sci. USA 2006, 103, 18314–18319. [Google Scholar] [CrossRef] [Green Version]
- Perkins, N.D. Integrating cell-signalling pathways with NF-kappaB and IKK function. Nat. Rev. Mol. Cell Biol. 2007, 8, 49–62. [Google Scholar] [CrossRef]
- Bretones, G.; Delgado, M.D.; Leon, J. Myc and cell cycle control. Biochim. Biophys. Acta. 2015, 1849, 506–516. [Google Scholar] [CrossRef] [PubMed]
- Lin, W.; Kemper, A.; McCarthy, K.D.; Pytel, P.; Wang, J.P.; Campbell, I.L.; Utset, M.F.; Popko, B. Interferon-gamma induced medulloblastoma in the developing cerebellum. J. Neurosci. 2004, 24, 10074–10083. [Google Scholar] [CrossRef] [PubMed]
- Meister, N.; Shalaby, T.; Von Bueren, A.O.; Rivera, P.; Patti, R.; Oehler, C.; Pruschy, M.; Grotzer, M.A. Interferon-gamma mediated up-regulation of caspase-8 sensitizes medulloblastoma cells to radio- and chemotherapy. Eur. J. Cancer. 2007, 43, 1833–1841. [Google Scholar] [CrossRef] [PubMed]
- Laner-Plamberger, S.; Wolff, F.; Kaser-Eichberger, A.; Swierczynski, S.; Hauser-Kronberger, C.; Frischauf, A.M.; Eichberger, T. Hedgehog/GLI signaling activates suppressor of cytokine signaling 1 (SOCS1) in epidermal and neural tumor cells. PLoS ONE 2013, 8, e75317. [Google Scholar] [CrossRef] [PubMed]
- Schmitt, M.J.; Philippidou, D.; Reinsbach, S.E.; Margue, C.; Wienecke-Baldacchino, A.; Nashan, D.; Behrmann, I.; Kreis, S. Interferon-gamma-induced activation of Signal Transducer and Activator of Transcription 1 (STAT1) up-regulates the tumor suppressing microRNA-29 family in melanoma cells. Cell Commun. Signal. 2012, 10, 41. [Google Scholar] [CrossRef] [PubMed]
- Ahmed, M.; Cheng, M.; Zhao, Q.; Goldgur, Y.; Cheal, S.M.; Guo, H.F.; Larson, S.M.; Cheung, N.K.V. Humanized Affinity-matured Monoclonal Antibody 8H9 Has Potent Antitumor Activity and Binds to FG Loop of Tumor Antigen B7-H3. J. Biol. Chem. 2015, 290, 30018–30029. [Google Scholar] [CrossRef] [Green Version]
© 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
Share and Cite
Purvis, I.J.; Avilala, J.; Guda, M.R.; Venkataraman, S.; Vibhakar, R.; Tsung, A.J.; Velpula, K.K.; Asuthkar, S. Role of MYC-miR-29-B7-H3 in Medulloblastoma Growth and Angiogenesis. J. Clin. Med. 2019, 8, 1158. https://doi.org/10.3390/jcm8081158
Purvis IJ, Avilala J, Guda MR, Venkataraman S, Vibhakar R, Tsung AJ, Velpula KK, Asuthkar S. Role of MYC-miR-29-B7-H3 in Medulloblastoma Growth and Angiogenesis. Journal of Clinical Medicine. 2019; 8(8):1158. https://doi.org/10.3390/jcm8081158
Chicago/Turabian StylePurvis, Ian J., Janardhan Avilala, Maheedhara R. Guda, Sujatha Venkataraman, Rajeev Vibhakar, Andrew J. Tsung, Kiran K. Velpula, and Swapna Asuthkar. 2019. "Role of MYC-miR-29-B7-H3 in Medulloblastoma Growth and Angiogenesis" Journal of Clinical Medicine 8, no. 8: 1158. https://doi.org/10.3390/jcm8081158