Abstract
Aim:
To investigate whether human multiple myeloma (MM) cells secrete microvesicles (MVs) and whether the MVs secreted from MM cells (MM-MVs) promote angiogenesis.
Methods:
RPMI8226 human MM cells and EA.hy926 human umbilical vein cells were used. MVs isolated from RPMI8226 cells were characterized under laser confocal microscopy, electron microscopy and with flow cytometry. The fusion of MM-MVs and EA.hy926 cells was studied under confocal microscopy, and the transfer of CD138 to EA.hy926 cells was demonstrated with flow cytometry. The proliferation, invasion and tube formation of EA.hy926 cells in vitro were evaluated using MTT, transwell migration and tube formation assays, respectively. The vasculization of EA.hy926 cells in vivo was studied using Matrigel plug assay. The expression of IL-6 and VEGF was analyzed with PCR and ELISA.
Results:
MM-MVs from the RPMI 8226 cells had the characteristic cup-shape with diameter of 100–1000 nm. Most of the MM-MVs expressed phosphatidylserine and the myeloma cell marker CD138, confirming that they were derived from myeloma cells. After added to EA.hy926 cells, the MM-MVs transferred CD138 to the endothelial cells and significantly stimulated the endothelial cells to proliferate, invade, secrete IL-6 and VEGF, two key angiogenic factors of myeloma, and form tubes in vitro and in vivo.
Conclusion:
Our results confirm the presence of MVs in MM cells and support the idea that MM-MVs are newfound mediators for myeloma angiogenesis and may serve as a therapeutic target to treat MM.
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References
Strobeck M . Multiple myeloma therapies. Nat Rev Drug Discov 2007; 6: 181–2.
Singh AV, Bandi M, Raje N, Richardson P, Palladino MA, Chauhan D, et al. A novel vascular disrupting agent plinabulin triggers JNK-mediated apoptosis and inhibits angiogenesis in multiple myeloma cells. Blood 2011; 117: 5692–700.
Podar K, Chauhan D, Anderson KC . Bone marrow microenvironment and the identification of new targets for myeloma therapy. Leukemia 2008; 23: 10–24.
Purushothaman A, Uyama T, Kobayashi F, Yamada S, Sugahara K, Rapraeger AC, et al. Heparanase-enhanced shedding of syndecan-1 by myeloma cells promotes endothelial invasion and angiogenesis. Blood 2010; 115: 2449–57.
Hose D, Moreaux J, Meissner T, Seckinger A, Goldschmidt H, Benner A, et al. Induction of angiogenesis by normal and malignant plasma cells. Blood 2009; 114: 128–43.
Van der Pol E, Boing AN, Harrison P, Sturk A, Nieuwland R . Classification, functions, and clinical relevance of extracellular vesicles. Pharmacol Rev 2012; 64: 676–705.
Szczepanski MJ, Szajnik M, Welsh A, Whiteside TL, Boyiadzis M . Blast-derived microvesicles in sera from patients with acute myeloid leukemia suppress natural killer cell function via membrane-associated transforming growth factor-1. Haematologica 2011; 96: 1302–9.
Al-Nedawi K, Meehan B, Rak J . Microvesicles: Messengers and mediators of tumor progression. Cell Cycle 2009; 8: 2014–8.
Al-Nedawi K, Meehan B, Micallef J, Lhotak V, May L, Guha A, et al. Intercellular transfer of the oncogenic receptor EGFRvIII by microvesicles derived from tumour cells. Nat Cell Biol 2008; 10: 619–24.
Ratajczak J, Miekus K, Kucia M, Zhang J, Reca R, Dvorak P, et al. Embryonic stem cell-derived microvesicles reprogram hematopoietic progenitors: evidence for horizontal transfer of mRNA and protein delivery. Leukemia 2006; 20: 847–56.
Bebawy M, Combes V, Lee E, Jaiswal R, Gong J, Bonhoure A, et al. Membrane microparticles mediate transfer of P-glycoprotein to drug sensitive cancer cells. Leukemia 2009; 23: 1643–9.
Wysoczynski M, Ratajczak MZ . Lung cancer secreted microvesicles: Underappreciated modulators of microenvironment in expanding tumors. Int J Cancer 2009; 125: 1595–603.
Ghosh AK, Secreto CR, Knox TR, Ding W, Mukhopadhyay D, Kay NE . Circulating microvesicles in B-cell chronic lymphocytic leukemia can stimulate marrow stromal cells: implications for disease progression. Blood 2010; 115: 1755–64.
Kawamoto T, Ohga N, Akiyama K, Hirata N, Kitahara S, Maishi N, et al. Tumor-derived microvesicles induce proangiogenic phenotype in endothelial cells via endocytosis. PLoS One 2012; 7: e34045.
Antonyak MA, Li B, Boroughs LK, Johnson JL, Druso JE, Bryant KL, et al. Cancer cell-derived microvesicles induce transformation by transferring tissue transglutaminase and fibronectin to recipient cells. Proc Natl Acad Sci U S A 2011; 108: 4852–7.
Stoorvogel W . Functional transfer of microRNA by exosomes. Blood 2012; 119: 646–8.
Jaiswal R, Gong J, Sambasivam S, Combes V, Mathys JM, Davey R, et al. Microparticle-associated nucleic acids mediate trait dominance in cancer. FASEB J 2012; 26: 420–29.
D'Souza-Schorey C, Clancy JW . Tumor-derived microvesicles: shedding light on novel microenvironment modulators and prospective cancer biomarkers. Genes Dev 2012; 26: 1287–99.
Umezu T, Ohyashiki K, Kuroda M, Ohyashiki JH . Leukemia cell to endothelial cell communication via exosomal miRNAs. Oncogene 2013; 32: 2747–55.
Thompson CA, Purushothaman A, Ramani VC, Vlodavsky I, Sanderson RD . Heparanase regulates secretion, composition, and function of tumor cell-derived exosomes. J Biol Chem 2013; 288: 10093–9.
Mineo M, Garfield SH, Taverna S, Flugy A, De Leo G, Alessandro R, et al. Exosomes released by K562 chronic myeloid leukemia cells promote angiogenesis in a src-dependent fashion. Angiogenesis 2011; 15: 33–45.
Grange C, Tapparo M, Collino F, Vitillo L, Damasco C, Deregibus MC, et al. Microvesicles released from human renal cancer stem cells stimulate angiogenesis and formation of lung premetastatic niche. Cancer Res 2011; 71: 5346–56.
Millimaggi D, Mari M, D'Ascenzo S, Carosa E, Jannini EA, Zucker S, et al. Tumor vesicle-associated CD147 modulates the angiogenic capability of endothelial cells. Neoplasia 2007; 9: 349–57.
Castellana D, Zobairi F, Martinez MC, Panaro MA, Mitolo V, Freyssinet JM, et al. Membrane microvesicles as actors in the establishment of a favorable prostatic tumoral niche: a role for activated fibroblasts and CX3CL1-CX3CR1 axis. Cancer Res 2009; 69: 785–93.
Chai C, Zheng S, Feng J, Wu X, Yang J, Wei M . A novel method for establishment and characterization of extrahepatic bile duct epithelial cells from mice. In Vitro Cell Dev Biol Anim 2010; 46: 820–3.
Pang X, Yi T, Yi Z, Cho SG, Qu W, Pinkaew D, et al. Morelloflavone, a biflavonoid, inhibits tumor angiogenesis by targeting Rho GTPases and extracellular signal-regulated kinase signaling pathways. Cancer Res 2009; 69: 518–25.
Beyer C, Pisetsky DS . The role of microparticles in the pathogenesis of rheumatic diseases. Nat Rev Rheumatol 2010; 6: 21–9.
Matsui W, Huff CA, Wang Q, Malehorn MT, Barber J, Tanhehco Y, et al. Characterization of clonogenic multiple myeloma cells. Blood 2004; 103: 2332–6.
Pap E, Pállinger É, Falus A . The role of membrane vesicles in tumorigenesis. Crit Rev Oncol Hematol 2011; 79: 213–23.
Dankbar B, Padró T, Leo R, Feldmann B, Kropff M, Mesters RM, et al. Vascular endothelial growth factor and interleukin-6 in paracrine tumor-stromal cell interactions in multiple myeloma. Blood 2000; 95: 2630–6.
Fan F, Schimming A, Jaeger D, Podar K . Targeting the tumor microenvironment: focus on angiogenesis. J Oncol 2012; 2012: 1–16.
Nguyen DH, Oketch-Rabah HA, Illa-Bochaca I, Geyer FC, Reis-Filho JS, Mao JH, et al. Radiation acts on the microenvironment to affect breast carcinogenesis by distinct mechanisms that decrease cancer latency and affect tumor type. Cancer Cell 2011; 19: 640–51.
Li B, Antonyak MA, Zhang J, Cerione RA . RhoA triggers a specific signaling pathway that generates transforming microvesicles in cancer cells. Oncogene 2012; 31: 4740–9.
Roccaro AM, Sacco A, Maiso P, Azab AK, Tai YT, Reagan M, et al. BM mesenchymal stromal cell-derived exosomes facilitate multiple myeloma progression. J Clin Invest 2013; 123: 1542–55.
Camussi G, Deregibus MC, Bruno S, Grange C, Fonsato V, Tetta C . Exosome/microvesicle-mediated epigenetic reprogramming of cells. Am J Cancer Res 2011; 1: 98–110.
Heijnen HFG, Schiel AE, Fijnheer R, Geuze HJ, Sixma JJ . Activated platelets release two types of membrane vesicles: microvesicles by surface shedding and exosomes derived from exocytosis of multivesicular bodies and alpha-granules. Blood 1999; 94: 3791–9.
Gonzalez G, Vituret C, Di Pietro A, Chanson M, Boulanger P, Hong SS . Microparticle-mediated transfer of the viral receptors CAR and CD46, and the CFTR channel in a CHO cell model confers new functions to target cells. PLoS One 2012; 7: e52326.
Khotskaya YB, Dai Y, Ritchie JP, MacLeod V, Yang Y, Zinn K, et al. Syndecan-1 is required for robust growth, vascularization, and metastasis of myeloma tumors in vivo. J Biol Chem 2009; 284: 26085–95.
Yang Y, Yaccoby S, Liu W, Langford JK, Pumphrey CY, Theus A, et al. Soluble syndecan-1 promotes growth of myeloma tumors in vivo. Blood 2002; 100: 610–7.
Ritchie JP, Ramani VC, Ren Y, Naggi A, Torri G, Casu B, et al. SST0001, a chemically modified heparin, inhibits myeloma growth and angiogenesis via disruption of the heparanase/syndecan-1 axis. Clin Cancer Res 2011; 17: 1382–93.
Kumar S, Witzig TE, Timm M, Haug J, Wellik L, Fonseca R, et al. Expression of VEGF and its receptors by myeloma cells. Leukemia 2003; 17: 2025–31.
Ria R, Todoerti K, Berardi S, Coluccia AML, De Luisi A, Mattioli M, et al. Gene expression profiling of bone marrow endothelial cells in patients with multiple myeloma. Clin Cancer Res 2009; 15: 5369–78.
De Luisi A, Ferrucci A, Coluccia AML, Ria R, Moschetta M, de Luca E, et al. Lenalidomide restrains motility and overangiogenic potential of bone marrow endothelial cells in patients with active multiple myeloma. Clin Cancer Res 2011; 17: 1935–46.
Peinado H, Lavotshkin S, Lyden D . The secreted factors responsible for pre-metastatic niche formation: old sayings and new thoughts. Semin Cancer Biol 2011; 21: 139–46.
Hergenreider E, Heydt S, Tréguer K, Boettger T, Horrevoets AJG, Zeiher AM, et al. Atheroprotective communication between endothelial cells and smooth muscle cells through miRNAs. Nat Cell Biol 2012; 14: 249–56.
Yang M, Chen J, Su F, Yu B, Su F, Lin L, et al. Microvesicles secreted by macrophages shuttle invasion-potentiating microRNAs into breast cancer cells. Mol Cancer 2011; 10: 117.
Balaj L, Lessard R, Dai L, Cho YJ, Pomeroy SL, Breakefield XO, et al. Tumour microvesicles contain retrotransposon elements and amplified oncogene sequences. Nat Commun 2011; 2: 180.
Piccin A, Murphy WG, Smith OP . Circulating microparticles: pathophysiology and clinical implications. Blood Rev 2007; 21: 157–71.
Shao H, Chung J, Balaj L, Charest A, Bigner DD, Carter BS, et al. Protein typing of circulating microvesicles allows real-time monitoring of glioblastoma therapy. Nat Med 2012; 18: 1835–40.
Acknowledgements
This research was supported by grants from the National Natural Science Foundation of China (81272624 and 81071943). We would like to thank the Flow Cytometry Lab of Tongji Medical College for the detection and the analysis of the surface markers of the MM-MVs.
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Liu, Y., Zhu, Xj., Zeng, C. et al. Microvesicles secreted from human multiple myeloma cells promote angiogenesis. Acta Pharmacol Sin 35, 230–238 (2014). https://doi.org/10.1038/aps.2013.141
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DOI: https://doi.org/10.1038/aps.2013.141
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