Abstract
Semaphorins are implicated in glioma progression, although little is known about the underlying mechanisms. We have reported plexin-B3 expression in human gliomas, which upon stimulation by Sema5A causes significant inhibition of cell migration and invasion. The concomitant inactivation of Rac1 is of mechanistic importance because forced expression of constitutively active Rac1 abolishes these inhibitory effects. Furthermore, Sema5A induces prominent cell collapse and ramification of processes reminiscent of astrocytic morphology, which temporally associate with extensive disassembly of actin stress fibers and disruption of focal adhesions, followed by accumulation of actin patches in protrusions. Mechanistically, Sema5A induces transient protein kinase C (PKC) phosphorylation of fascin-1, which can reduce its actin-binding/bundling activities and temporally parallels its translocation from cell body to extending processes. PKC inhibition or fascin-1 knockdown is sufficient to abrogate Sema5A-induced morphological differentiation, whereas the process is hastened by forced expression of fascin-1. Intriguingly, Sema5A induces re-expression of glial fibrillary acidic protein (GFAP), which when silenced restricts differentiation of glioma cells to bipolar instead of multipolar morphology. Therefore, we hypothesize complementary functions of fascin-1 and GFAP in the early and late phases of Sema5A-induced astrocytic differentiation of gliomas, respectively. In summary, Sema5A and plexin-B3 impede motility but promote differentiation of human gliomas. These effects are plausibly compromised in high-grade human astrocytomas in which Sema5A expression is markedly reduced, hence leading to infiltrative and anaplastic characteristics. This is evident by increased invasiveness of glioma cells when endogenous Sema5A is silenced. Therefore, Sema5A and plexin-B3 represent potential novel targets in counteracting glioma progression.
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References
Adams JC . (2004). Roles of fascin in cell adhesion and motility. Curr Opin Cell Biol 16: 590–596.
Anilkumar N, Parsons M, Monk R, Ng T, Adams JC . (2003). Interaction of fascin and protein kinase Calpha: a novel intersection in cell adhesion and motility. EMBO J 22: 5390–5402.
Artigiani S, Conrotto P, Fazzari P, Gilestro GF, Barberis D, Giordano S et al. (2004). Plexin-B3 is a functional receptor for semaphorin 5A. EMBO Rep 5: 710–714.
Bagci T, Wu JK, Pfannl R, Ilag LL, Jay DG . (2009). Autocrine semaphorin 3A signaling promotes glioblastoma dispersal. Oncogene 28: 3537–3550.
Balakrishnan A, Penachioni JY, Lamba S, Bleeker FE, Zanon C, Rodolfo M et al. (2009). Molecular profiling of the ‘plexinome’ in melanoma and pancreatic cancer. Hum Mutat 30: 1167–1174.
Banu N, Teichman J, Dunlap-Brown M, Villegas G, Tufro A . (2006). Semaphorin 3C regulates endothelial cell function by increasing integrin activity. FASEB J 20: 2150–2152.
Barberis D, Artigiani S, Casazza A, Corso S, Giordano S, Love CA et al. (2004). Plexin signaling hampers integrin-based adhesion, leading to Rho-kinase independent cell rounding, and inhibiting lamellipodia extension and cell motility. FASEB J 18: 592–594.
Basile JR, Castilho RM, Williams VP, Gutkind JS . (2006). Semaphorin 4D provides a link between axon guidance processes and tumor-induced angiogenesis. Proc Natl Acad Sci USA 103: 9017–9022.
Bellail AC, Hunter SB, Brat DJ, Tan C, Van Meir EG . (2004). Microregional extracellular matrix heterogeneity in brain modulates glioma cell invasion. Int J Biochem Cell Biol 36: 1046–1069.
Benard V, Bohl BP, Bokoch GM . (1999). Characterization of rac and cdc42 activation in chemoattractant-stimulated human neutrophils using a novel assay for active GTPases. J Biol Chem 274: 13198–13204.
Brun M, Coles JE, Monckton EA, Glubrecht DD, Bisgrove D, Godbout R . (2009). Nuclear factor I regulates brain fatty acid-binding protein and glial fibrillary acidic protein gene expression in malignant glioma cell lines. J Mol Biol 391: 282–300.
Castro-Rivera E, Ran S, Thorpe P, Minna JD . (2004). Semaphorin 3B (SEMA3B) induces apoptosis in lung and breast cancer, whereas VEGF165 antagonizes this effect. Proc Natl Acad Sci USA 101: 11432–11437.
Catalano A, Caprari P, Rodilossi S, Betta P, Castellucci M, Casazza A et al. (2004). Cross-talk between vascular endothelial growth factor and semaphorin-3A pathway in the regulation of normal and malignant mesothelial cell proliferation. FASEB J 18: 358–360.
Chan AY, Coniglio SJ, Chuang YY, Michaelson D, Knaus UG, Philips MR et al. (2005). Roles of the Rac1 and Rac3 GTPases in human tumor cell invasion. Oncogene 24: 7821–7829.
Chedotal A . (2007). Chemotropic axon guidance molecules in tumorigenesis. Prog Exp Tumor Res 39: 78–90.
Christensen C, Ambartsumian N, Gilestro G, Thomsen B, Comoglio P, Tamagnone L et al. (2005). Proteolytic processing converts the repelling signal Sema3E into an inducer of invasive growth and lung metastasis. Cancer Res 65: 6167–6177.
Coma S, Amin DN, Shimizu A, Lasorella A, Iavarone A, Klagsbrun M . (2010). Id2 promotes tumor cell migration and invasion through transcriptional repression of semaphorin 3F. Cancer Res 70: 3823–3832.
Conrotto P, Corso S, Gamberini S, Comoglio PM, Giordano S . (2004). Interplay between scatter factor receptors and B plexins controls invasive growth. Oncogene 23: 5131–5137.
Correa RG, Sasahara RM, Bengtson MH, Katayama ML, Salim AC, Brentani MM et al. (2001). Human semaphorin 6B [(HSA)SEMA6B], a novel human class 6 semaphorin gene: alternative splicing and all-trans-retinoic acid-dependent downregulation in glioblastoma cell lines. Genomics 73: 343–348.
Crowe DL . (2004). Overlapping functions of Ras and Rac GTPases in regulating cancer cell proliferation and invasion. Anticancer Res 24: 593–597.
Deck JH, Eng LF, Bigbee J, Woodcock SM . (1978). The role of glial fibrillary acidic protein in the diagnosis of central nervous system tumors. Acta Neuropathol 42: 183–190.
Deryugina EI, Bourdon MA, Luo GX, Reisfeld RA, Strongin A . (1997). Matrix metalloproteinase-2 activation modulates glioma cell migration. J Cell Sci 110: 2473–2482.
Dhanabal M, Wu F, Alvarez E, McQueeney KD, Jeffers M, MacDougall J et al. (2005). Recombinant semaphorin 6A-1 ectodomain inhibits in vivo growth factor and tumor cell line-induced angiogenesis. Cancer Biol Ther 4: 659–668.
Driessens MH, Hu H, Nobes CD, Self A, Jordens I, Goodman CS et al. (2001). Plexin-B semaphorin receptors interact directly with active Rac and regulate the actin cytoskeleton by activating Rho. Curr Biol 11: 339–344.
Driessens MH, Olivo C, Nagata K, Inagaki M, Collard JG . (2002). B plexins activate Rho through PDZ-RhoGEF. FEBS Lett 529: 168–172.
Duffy PE, Huang YY, Rapport MM . (1982). The relationship of glial fibrillary acidic protein to the shape, motility, and differentiation of human astrocytoma cells. Exp Cell Res 139: 145–157.
Elobeid A, Bongcam-Rudloff E, Westermark B, Nister M . (2000). Effects of inducible glial fibrillary acidic protein on glioma cell motility and proliferation. J Neurosci Res 60: 245–256.
Feldkamp MM, Lau N, Guha A . (1999). Growth inhibition of astrocytoma cells by farnesyl transferase inhibitors is mediated by a combination of anti-proliferative, pro-apoptotic and anti-angiogenic effects. Oncogene 18: 7514–7526.
Goldberg JL, Vargas ME, Wang JT, Mandemakers W, Oster SF, Sretavan DW et al. (2004). An oligodendrocyte lineage-specific semaphorin, Sema5A, inhibits axon growth by retinal ganglion cells. J Neurosci 24: 4989–4999.
Hilario JD, Rodino-Klapac LR, Wang C, Beattie CE . (2009). Semaphorin 5A is a bifunctional axon guidance cue for axial motoneurons in vivo. Dev Biol 326: 190–200.
Hu B, Guo P, Bar-Joseph I, Imanishi Y, Jarzynka MJ, Bogler O et al. (2007). Neuropilin-1 promotes human glioma progression through potentiating the activity of the HGF/SF autocrine pathway. Oncogene 26: 5577–5586.
Hu H, Marton TF, Goodman CS . (2001). Plexin B mediates axon guidance in Drosophila by simultaneously inhibiting active Rac and enhancing RhoA signaling. Neuron 32: 39–51.
Inagaki M, Gonda Y, Nishizawa K, Kitamura S, Sato C, Ando S et al. (1990). Phosphorylation sites linked to glial filament disassembly in vitro locate in a non-alpha-helical head domain. J Biol Chem 265: 4722–4729.
Iwadate Y, Sakaida T, Hiwasa T, Nagai Y, Ishikura H, Takiguchi M et al. (2004). Molecular classification and survival prediction in human gliomas based on proteome analysis. Cancer Res 64: 2496–2501.
Kantor DB, Chivatakarn O, Peer KL, Oster SF, Inatani M, Hansen MJ et al. (2004). Semaphorin 5A is a bifunctional axon guidance cue regulated by heparan and chondroitin sulfate proteoglycans. Neuron 44: 961–975.
Karayan-Tapon L, Wager M, Guilhot J, Levillain P, Marquant C, Clarhaut J et al. (2008). Semaphorin, neuropilin and VEGF expression in glial tumours: SEMA3G, a prognostic marker? Br J Cancer 99: 1153–1160.
Kureishy N, Sapountzi V, Prag S, Anilkumar N, Adams JC . (2002). Fascins, and their roles in cell structure and function. Bioessays 24: 350–361.
Kusy S, Nasarre P, Chan D, Potiron V, Meyronet D, Gemmill RM et al. (2005). Selective suppression of in vivo tumorigenicity by semaphorin SEMA3F in lung cancer cells. Neoplasia 7: 457–465.
Li X, Lee AY . (2010). Semaphorin 5A and plexin-B3 inhibit human glioma cell motility through RhoGDIalpha-mediated inactivation of Rac1 GTPase. J Biol Chem 285: 32436–32445.
Lu TP, Tsai MH, Lee JM, Hsu CP, Chen PC, Lin CW et al. (2010). Identification of a novel biomarker, SEMA5A, for non-small cell lung carcinoma in nonsmoking women. Cancer Epidemiol Biomarkers Prev 19: 2590–2597.
McDonough WS, Tran NL, Berens ME . (2005). Regulation of glioma cell migration by serine-phosphorylated P311. Neoplasia 7: 862–872.
Mondal S, Dirks P, Rutka JT . (2009). Immunolocalization of fascin, an actin-bundling protein and glial fibrillary acidic protein in human astrocytoma cells. Brain Pathol 20: 190–199.
Murakami M, Goto S, Yoshikawa M, Goto T, Hamasaki T, Rutka JT et al. (2001). The invasive features of glial and non-central nervous system tumor cells are different on organotypic brain slices from newborn rats. Int J Oncol 18: 721–727.
Nakada M, Drake KL, Nakada S, Niska JA, Berens ME . (2006). Ephrin-B3 ligand promotes glioma invasion through activation of Rac1. Cancer Res 66: 8492–8500.
Nasarre C, Koncina E, Labourdette G, Cremel G, Roussel G, Aunis D et al. (2009). Neuropilin-2 acts as a modulator of Sema3A-dependent glioma cell migration. Cell Adh Migr 3: 383–389.
Neufeld G, Shraga-Heled N, Lange T, Guttmann-Raviv N, Herzog Y, Kessler O . (2005). Semaphorins in cancer. Front Biosci 10: 751–760.
Oinuma I, Ishikawa Y, Katoh H, Negishi M . (2004). The semaphorin 4D receptor plexin-B1 is a GTPase activating protein for R-Ras. Science 305: 862–865.
Oinuma I, Katoh H, Negishi M . (2006). Semaphorin 4D/plexin-B1-mediated R-Ras GAP activity inhibits cell migration by regulating beta(1) integrin activity. J Cell Biol 173: 601–613.
Oster SF, Bodeker MO, He F, Sretavan DW . (2003). Invariant Sema5A inhibition serves an ensheathing function during optic nerve development. Development 130: 775–784.
Pan G, Lv H, Ren H, Wang Y, Liu Y, Jiang H et al. (2010). Elevated expression of semaphorin 5A in human gastric cancer and its implication in carcinogenesis. Life Sci 86: 139–144.
Pan GQ, Ren HZ, Zhang SF, Wang XM, Wen JF . (2009). Expression of semaphorin 5A and its receptor plexin B3 contributes to invasion and metastasis of gastric carcinoma. World J Gastroenterol 15: 2800–2804.
Restrepo A, Smith CA, Agnihotri S, Shekarforoush M, Kongkham PN, Seol HJ et al. (2010). Epigenetic regulation of glial fibrillary acidic protein by DNA methylation in human malignant gliomas. Neuro Oncol 13: 42–50.
Rich JN, Hans C, Jones B, Iversen ES, McLendon RE, Rasheed BK et al. (2005). Gene expression profiling and genetic markers in glioblastoma survival. Cancer Res 65: 4051–4058.
Rickman DS, Bobek MP, Misek DE, Kuick R, Blaivas M, Kurnit DM et al. (2001). Distinctive molecular profiles of high-grade and low-grade gliomas based on oligonucleotide microarray analysis. Cancer Res 61: 6885–6891.
Ridley AJ, Hall A . (1992). The small GTP-binding protein rho regulates the assembly of focal adhesions and actin stress fibers in response to growth factors. Cell 70: 389–399.
Rieger J, Wick W, Weller M . (2003). Human malignant glioma cells express semaphorins and their receptors, neuropilins and plexins. Glia 42: 379–389.
Roberts DD . (1996). Regulation of tumor growth and metastasis by thrombospondin-1. FASEB J 10: 1183–1191.
Rutka JT, Hubbard SL, Fukuyama K, Matsuzawa K, Dirks PB, Becker LE . (1994). Effects of antisense glial fibrillary acidic protein complementary DNA on the growth, invasion, and adhesion of human astrocytoma cells. Cancer Res 54: 3267–3272.
Rutka JT, Murakami M, Dirks PB, Hubbard SL, Becker LE, Fukuyama K et al. (1997). Role of glial filaments in cells and tumors of glial origin: a review. J Neurosurg 87: 420–430.
Rutka JT, Smith SL . (1993). Transfection of human astrocytoma cells with glial fibrillary acidic protein complementary DNA: analysis of expression, proliferation, and tumorigenicity. Cancer Res 53: 3624–3631.
Sadanandam A, Varney ML, Kinarsky L, Ali H, Mosley RL, Singh RK . (2007). Identification of functional cell adhesion molecules with a potential role in metastasis by a combination of in vivo phage display and in silico analysis. OMICS 11: 41–57.
Sadanandam A, Varney ML, Singh S, Ashour AE, Moniaux N, Deb S et al. (2010). High gene expression of semaphorin 5A in pancreatic cancer is associated with tumor growth, invasion and metastasis. Int J Cancer 127: 1373–1383.
Serini G, Valdembri D, Zanivan S, Morterra G, Burkhardt C, Caccavari F et al. (2003). Class 3 semaphorins control vascular morphogenesis by inhibiting integrin function. Nature 424: 391–397.
Swiercz JM, Kuner R, Offermanns S . (2004). Plexin-B1/RhoGEF-mediated RhoA activation involves the receptor tyrosine kinase ErbB-2. J Cell Biol 165: 869–880.
Swiercz JM, Worzfeld T, Offermanns S . (2008). ErbB-2 and met reciprocally regulate cellular signaling via plexin-B1. J Biol Chem 283: 1893–1901.
Tamagnone L, Artigiani S, Chen H, He Z, Ming GI, Song H et al. (1999). Plexins are a large family of receptors for transmembrane, secreted, and GPI-anchored semaphorins in vertebrates. Cell 99: 71–80.
Walzer T, Galibert L, De Smedt T . (2005). Poxvirus semaphorin A39R inhibits phagocytosis by dendritic cells and neutrophils. Eur J Immunol 35: 391–398.
Webb DJ, Brown CM, Horwitz AF . (2003). Illuminating adhesion complexes in migrating cells: moving toward a bright future. Curr Opin Cell Biol 15: 614–620.
Wick W, Platten M, Weller M . (2001). Glioma cell invasion: regulation of metalloproteinase activity by TGF-beta. J Neurooncol 53: 177–185.
Wong CC, Wong CM, Ko FC, Chan LK, Ching YP, Yam JW et al. (2008). Deleted in liver cancer 1 (DLC1) negatively regulates Rho/ROCK/MLC pathway in hepatocellular carcinoma. PLoS ONE 3: e2779.
Woodhouse EC, Fisher A, Bandle RW, Bryant-Greenwood B, Charboneau L, Petricoin III EF et al. (2003). Drosophila screening model for metastasis: semaphorin 5c is required for l(2)gl cancer phenotype. Proc Natl Acad Sci USA 100: 11463–11468.
Yamashiro S, Yamakita Y, Ono S, Matsumura F . (1998). Fascin, an actin-bundling protein, induces membrane protrusions and increases cell motility of epithelial cells. Mol Biol Cell 9: 993–1006.
Zhuge Y, Xu J . (2001). Rac1 mediates type I collagen-dependent MMP-2 activation. Role in cell invasion across collagen barrier. J Biol Chem 276: 16248–16256.
Acknowledgements
We are indebted to Dr David Srevatan for providing the Sema5A-Fc expression construct and to Dr Danijela Vignijevic for the pEGFP-fascin plasmid. This work was supported by research grants from the National University of Singapore and the Biomedical Research Council of Singapore.
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Li, X., Law, J. & Lee, A. Semaphorin 5A and plexin-B3 regulate human glioma cell motility and morphology through Rac1 and the actin cytoskeleton. Oncogene 31, 595–610 (2012). https://doi.org/10.1038/onc.2011.256
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