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STIM1 and Orai1 mediate CRAC channel activity and are essential for human glioblastoma invasion

  • Ion Channels, Receptors and Transporters
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Abstract

The Ca2+ sensor stromal interacting molecule 1 (STIM1) and the Ca2+ channel Orai1 mediate the ubiquitous store-operated Ca2+ entry (SOCE) pathway activated by depletion of internal Ca2+ stores and mediated through the highly Ca2+-selective, Ca2+ release-activated Ca2+ (CRAC) current. Furthermore, STIM1 and Orai1, along with Orai3, encode store-independent Ca2+ currents regulated by either arachidonate or its metabolite, leukotriene C4. Orai channels are emerging as important contributors to numerous cell functions, including proliferation, migration, differentiation, and apoptosis. Recent studies suggest critical involvement of STIM/Orai proteins in controlling the development of several cancers, including malignancies of the breast, prostate, and cervix. Here, we quantitatively compared the magnitude of SOCE and the expression levels of STIM1 and Orai1 in non-malignant human primary astrocytes (HPA) and in primary human cell lines established from surgical samples of the brain tumor glioblastoma multiforme (GBM). Using Ca2+ imaging, patch-clamp electrophysiology, pharmacological reagents, and gene silencing, we established that in GBM cells, SOCE and CRAC are mediated by STIM1 and Orai1. We further found that GBM cells show upregulation of SOCE and increased Orai1 levels compared to HPA. The functional significance of SOCE was evaluated by studying the effects of STIM1 and Orai1 knockdown on cell proliferation and invasion. Utilizing Matrigel assays, we demonstrated that in GBM, but not in HPA, downregulation of STIM1 and Orai1 caused a dramatic decrease in cell invasion. In contrast, the effects of STIM1 and Orai1 knockdown on GBM cell proliferation were marginal. Overall, these results demonstrate that STIM1 and Orai1 encode SOCE and CRAC currents and control invasion of GBM cells. Our work further supports the potential use of channels contributed by Orai isoforms as therapeutic targets in cancer.

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References

  1. Abdullaev IF, Bisaillon JM, Potier M, Gonzalez JC, Motiani RK, Trebak M (2008) Stim1 and Orai1 mediate CRAC currents and store-operated calcium entry important for endothelial cell proliferation. Circ Res 103(11):1289–1299. doi:10.1161/01.RES.0000338496.95579.56

    Article  PubMed  CAS  Google Scholar 

  2. Abdullaev IF, Rudkouskaya A, Mongin AA, Kuo YH (2010) Calcium-activated potassium channels BK and IK1 are functionally expressed in human gliomas but do not regulate cell proliferation. PLoS One 5(8):e12304. doi:10.1371/journal.pone.0012304

    Article  PubMed  Google Scholar 

  3. Barnholtz-Sloan JS, Sloan AE, Schwartz AG (2003) Relative survival rates and patterns of diagnosis analyzed by time period for individuals with primary malignant brain tumor, 1973–1997. J Neurosurg 99(3):458–466. doi:10.3171/jns.2003.99.3.0458

    Article  PubMed  Google Scholar 

  4. Behin A, Hoang-Xuan K, Carpentier AF, Delattre JY (2003) Primary brain tumours in adults. Lancet 361(9354):323–331. doi:10.1016/S0140-6736(03)12328-8

    Article  PubMed  Google Scholar 

  5. Berridge MJ (1993) Inositol trisphosphate and calcium signalling. Nature 361(6410):315–325

    Article  PubMed  CAS  Google Scholar 

  6. Bird GS, DeHaven WI, Smyth JT, Putney JW Jr (2008) Methods for studying store-operated calcium entry. Methods 46(3):204–212. doi:10.1016/j.ymeth.2008.09.009

    Article  PubMed  CAS  Google Scholar 

  7. Bisaillon JM, Motiani RK, Gonzalez-Cobos JC, Potier M, Halligan KE, Alzawahra WF, Barroso M, Singer HA, Jourd’heuil D, Trebak M (2010) Essential role for STIM1/Orai1-mediated calcium influx in PDGF-induced smooth muscle migration. Am j physiol Cell physiol 298(5):C993–C1005

    Article  PubMed  CAS  Google Scholar 

  8. Chen YF, Chiu WT, Chen YT, Lin PY, Huang HJ, Chou CY, Chang HC, Tang MJ, Shen MR (2011) Calcium store sensor stromal-interaction molecule 1-dependent signaling plays an important role in cervical cancer growth, migration, and angiogenesis. Proc Natl Acad Sci U S A 108(37):15225–15230. doi:10.1073/pnas.1103315108

    Article  PubMed  CAS  Google Scholar 

  9. Derler I, Schindl R, Fritsch R, Romanin C (2012) Gating and permeation of Orai channels. Front biosci j virtual libr 17:1304–1322

    Article  CAS  Google Scholar 

  10. Faouzi M, Hague F, Potier M, Ahidouch A, Sevestre H, Ouadid-Ahidouch H (2011) Down-regulation of Orai3 arrests cell-cycle progression and induces apoptosis in breast cancer cells but not in normal breast epithelial cells. J Cell Physiol 226(2):542–551. doi:10.1002/jcp.22363

    Article  PubMed  CAS  Google Scholar 

  11. Faouzi M, Kischel P, Hague F, Ahidouch A, Benzerdjeb N, Sevestre H, Penner R, Ouadid-Ahidouch H (2012) ORAI3 silencing alters cell proliferation and cell cycle progression via c-myc pathway in breast cancer cells. Biochim Biophys Acta. doi:10.1016/j.bbamcr.2012.12.009

  12. Feng M, Grice DM, Faddy HM, Nguyen N, Leitch S, Wang Y, Muend S, Kenny PA, Sukumar S, Roberts-Thomson SJ, Monteith GR, Rao R (2010) Store-independent activation of Orai1 by SPCA2 in mammary tumors. Cell 143(1):84–98. doi:10.1016/j.cell.2010.08.040

    Article  PubMed  CAS  Google Scholar 

  13. Flourakis M, Lehen’kyi V, Beck B, Raphael M, Vandenberghe M, Abeele FV, Roudbaraki M, Lepage G, Mauroy B, Romanin C, Shuba Y, Skryma R, Prevarskaya N (2010) Orai1 contributes to the establishment of an apoptosis-resistant phenotype in prostate cancer cells. Cell death dis 1:e75. doi:10.1038/cddis.2010.52

    Article  PubMed  CAS  Google Scholar 

  14. Furnari FB, Fenton T, Bachoo RM, Mukasa A, Stommel JM, Stegh A, Hahn WC, Ligon KL, Louis DN, Brennan C, Chin L, DePinho RA, Cavenee WK (2007) Malignant astrocytic glioma: genetics, biology, and paths to treatment. Genes Dev 21(21):2683–2710. doi:10.1101/gad.1596707

    Article  PubMed  CAS  Google Scholar 

  15. Gonzalez-Cobos JC, Zhang X, Zhang W, Ruhle BC, Motiani RK, Schindl R, Muik M, Spinelli AM, Bisaillon JM, Shinde AV, Fahrner M, Singer HA, Matrougui K, Barroso M, Romanin C, Trebak M (2013) Store-independent Orai1/3 channels activated by intracrine leukotriene C4: role in neointimal hyperplasia. Circ Res. doi:10.1161/CIRCRESAHA.111.300220

  16. Hogan PG, Rao A (2007) Dissecting ICRAC, a store-operated calcium current. Trends biochem sci 32(5):235–245

    Article  PubMed  CAS  Google Scholar 

  17. Hoth M, Penner R (1992) Depletion of intracellular calcium stores activates a calcium current in mast cells. Nature 355(6358):353–356

    Article  PubMed  CAS  Google Scholar 

  18. Liu H, Hughes JD, Rollins S, Chen B, Perkins E (2011) Calcium entry via ORAI1 regulates glioblastoma cell proliferation and apoptosis. Exp Mol Pathol 91(3):753–760. doi:10.1016/j.yexmp.2011.09.005

    Article  PubMed  CAS  Google Scholar 

  19. McAndrew D, Grice DM, Peters AA, Davis FM, Stewart T, Rice M, Smart CE, Brown MA, Kenny PA, Roberts-Thomson SJ, Monteith GR (2011) ORAI1-mediated calcium influx in lactation and in breast cancer. Mol Cancer Ther 10(3):448–460. doi:10.1158/1535-7163.MCT-10-0923

    Article  PubMed  CAS  Google Scholar 

  20. Mignen O, Thompson JL, Shuttleworth TJ (2007) STIM1 regulates Ca2+ entry via arachidonate-regulated Ca2+-selective (ARC) channels without store depletion or translocation to the plasma membrane. J Physiol 579(Pt 3):703–715

    Article  PubMed  CAS  Google Scholar 

  21. Mignen O, Thompson JL, Shuttleworth TJ (2008) Both Orai1 and Orai3 are essential components of the arachidonate-regulated Ca2+-selective (ARC) channels. J Physiol 586(1):185–195

    Article  PubMed  CAS  Google Scholar 

  22. Monteith GR, McAndrew D, Faddy HM, Roberts-Thomson SJ (2007) Calcium and cancer: targeting Ca2+ transport. Nat Rev Cancer 7(7):519–530

    Article  PubMed  CAS  Google Scholar 

  23. Motiani RK, Abdullaev IF, Trebak M (2010) A novel native store-operated calcium channel encoded by Orai3: selective requirement of Orai3 versus Orai1 in estrogen receptor-positive versus estrogen receptor-negative breast cancer cells. J Biol Chem 285(25):19173–19183

    Article  PubMed  CAS  Google Scholar 

  24. Motiani RK, Zhang X, Harmon KE, Keller RS, Matrougui K, Bennett JA, Trebak M (2013) Orai3 is an estrogen receptor alpha-regulated Ca2+ channel that promotes tumorigenesis. FASEB j off publ Fed Am Soc Exp Biol 27(1):63–75. doi:10.1096/fj.12-213801

    CAS  Google Scholar 

  25. Muik M, Schindl R, Fahrner M, Romanin C (2012) Ca(2+) release-activated Ca(2+) (CRAC) current, structure, and function. Cell mol life sci CMLS 69(24):4163–4176. doi:10.1007/s00018-012-1072-8

    Article  CAS  Google Scholar 

  26. Parekh AB, Putney JW Jr (2005) Store-operated calcium channels. Physiol Rev 85(2):757–810. doi:10.1152/physrev.00057.2003

    Article  PubMed  CAS  Google Scholar 

  27. Potier M, Gonzalez JC, Motiani RK, Abdullaev IF, Bisaillon JM, Singer HA, Trebak M (2009) Evidence for STIM1- and Orai1-dependent store-operated calcium influx through ICRAC in vascular smooth muscle cells: role in proliferation and migration. FASEB j off publ Fed Am Soc Exp Biol 23(8):2425–2437. doi:10.1096/fj.09-131128

    CAS  Google Scholar 

  28. Potier M, Trebak M (2008) New developments in the signaling mechanisms of the store-operated calcium entry pathway. Pflugers Arch - Eur j physiol 457(2):405–415. doi:10.1007/s00424-008-0533-2

    Article  CAS  Google Scholar 

  29. Prakriya M, Feske S, Gwack Y, Srikanth S, Rao A, Hogan PG (2006) Orai1 is an essential pore subunit of the CRAC channel. Nature 443(7108):230–233

    Article  PubMed  CAS  Google Scholar 

  30. Prakriya M, Lewis RS (2001) Potentiation and inhibition of Ca(2+) release-activated Ca(2+) channels by 2-aminoethyldiphenyl borate (2-APB) occurs independently of IP(3) receptors. J Physiol 536(Pt 1):3–19

    Article  PubMed  CAS  Google Scholar 

  31. Prakriya M, Lewis RS (2002) Separation and characterization of currents through store-operated CRAC channels and Mg2+-inhibited cation (MIC) channels. J Gen Physiol 119(5):487–507

    Article  PubMed  CAS  Google Scholar 

  32. Prevarskaya N, Skryma R, Shuba Y (2011) Calcium in tumour metastasis: new roles for known actors. Nat Rev Cancer 11(8):609–618. doi:10.1038/nrc3105

    Article  PubMed  CAS  Google Scholar 

  33. Putney JW Jr (1986) A model for receptor-regulated calcium entry. Cell calcium 7(1):1–12

    Article  PubMed  CAS  Google Scholar 

  34. Putney JW Jr (1990) Capacitative calcium entry revisited. Cell calcium 11(10):611–624

    Article  PubMed  CAS  Google Scholar 

  35. Schwartzbaum JA, Fisher JL, Aldape KD, Wrensch M (2006) Epidemiology and molecular pathology of glioma. Nat Clin Pract Neurol 2(9):494–503. doi:10.1038/ncpneuro0289, quiz 491 p following 516

    Article  PubMed  Google Scholar 

  36. Scrideli CA, Carlotti CG Jr, Okamoto OK, Andrade VS, Cortez MA, Motta FJ, Lucio-Eterovic AK, Neder L, Rosemberg S, Oba-Shinjo SM, Marie SK, Tone LG (2008) Gene expression profile analysis of primary glioblastomas and non-neoplastic brain tissue: identification of potential target genes by oligonucleotide microarray and real-time quantitative PCR. J Neurooncol 88(3):281–291. doi:10.1007/s11060-008-9579-4

    Article  PubMed  CAS  Google Scholar 

  37. Shuttleworth TJ (2009) Arachidonic acid, ARC channels, and Orai proteins. Cell calcium 45:602–610

    Article  PubMed  CAS  Google Scholar 

  38. Spinelli AM, Gonzalez-Cobos JC, Zhang X, Motiani RK, Rowan S, Zhang W, Garrett J, Vincent PA, Matrougui K, Singer HA, Trebak M (2012) Airway smooth muscle STIM1 and Orai1 are upregulated in asthmatic mice and mediate PDGF-activated SOCE, CRAC currents, proliferation, and migration. Pflugers Arch - Eur j physiol 464(5):481–492. doi:10.1007/s00424-012-1160-5

    Article  CAS  Google Scholar 

  39. Suyama E, Wadhwa R, Kaur K, Miyagishi M, Kaul SC, Kawasaki H, Taira K (2004) Identification of metastasis-related genes in a mouse model using a library of randomized ribozymes. J Biol Chem 279(37):38083–38086. doi:10.1074/jbc.C400313200

    Article  PubMed  CAS  Google Scholar 

  40. Trebak M, Lemonnier L, Smyth JT, Vazquez G, Putney JW Jr (2007) Phospholipase C-coupled receptors and activation of TRPC channels. Handb Exp Pharmacol 179:593–614. doi:10.1007/978-3-540-34891-7_35

    Article  PubMed  CAS  Google Scholar 

  41. Yang S, Zhang JJ, Huang XY (2009) Orai1 and STIM1 are critical for breast tumor cell migration and metastasis. Cancer cell 15(2):124–134. doi:10.1016/j.ccr.2008.12.019

    Article  PubMed  CAS  Google Scholar 

  42. Zhang W, Halligan KE, Zhang X, Bisaillon JM, Gonzalez-Cobos JC, Motiani RK, Hu G, Vincent PA, Zhou J, Barroso M, Singer HA, Matrougui K, Trebak M (2011) Orai1-mediated I (CRAC) is essential for neointima formation after vascular injury. Circ Res 109(5):534–542

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This work was supported by NIH grant R01 HL097111 (to MT), AMC translational grant 206-465247 (to AAM and YHK), and in part by NIH grants R01 NS061953 (to AAM) and R01 HL095566 (to KM). The authors are grateful to Dr. M.G. Kaplitt for the gift of U251-MG cell line.

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Author notes

  1. Yu-Hung Kuo

    Present address: Department of Neurosurgery, Stony Brook University Medical Center, Stony Brook, NY, 11794, USA

Authors and Affiliations

  1. The College of Nanoscale Science and Engineering (CNSE), University at Albany, State University of New York, 257 Fuller Rd., Albany, NY, 12203, USA

    Rajender K. Motiani, Xuexin Zhang, Matthew M. Henkel & Mohamed Trebak

  2. The Center for Cardiovascular Sciences, Albany Medical College, Albany, NY, 12208, USA

    Rajender K. Motiani, Xuexin Zhang, Matthew M. Henkel & Mohamed Trebak

  3. The Center for Neuropharmacology and Neuroscience, Albany Medical College, 47 New Scotland Ave. (MC-136), Albany, NY, 12208, USA

    María C. Hyzinski-García, Iskandar F. Abdullaev & Alexander A. Mongin

  4. Division of Neurosurgery, Department of Surgery, Albany Medical College, Albany, NY, 12208, USA

    Yu-Hung Kuo

  5. Department of Physiology, Eastern Virginia Medical School, Norfolk, VA, 23501, USA

    Khalid Matrougui

  6. UNESCO-Regional Center for Biotechnology, New Delhi, India

    Rajender K. Motiani

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  1. Rajender K. Motiani
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  2. María C. Hyzinski-García
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Correspondence to Alexander A. Mongin or Mohamed Trebak.

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Alexander A. Mongin and Mohamed Trebak share the senior authorship.

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Motiani, R.K., Hyzinski-García, M.C., Zhang, X. et al. STIM1 and Orai1 mediate CRAC channel activity and are essential for human glioblastoma invasion. Pflugers Arch - Eur J Physiol 465, 1249–1260 (2013). https://doi.org/10.1007/s00424-013-1254-8

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  • DOI: https://doi.org/10.1007/s00424-013-1254-8

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