Journal of Korean Neurosurgical Society

The Korean Neurosurgical Society (대한신경외과학회)
권호 표지
DOI QR코드

DOI QR Code

Bacitracin Inhibits the Migration of U87-MG Glioma Cells via Interferences of the Integrin Outside-in Signaling Pathway

  • Li, Songyuan (Brain Tumor Research Laboratory and Department of Neurosurgery Chonnam National University Research Institute of Medical Sciences, Chonnam National University Hwasun Hopital and Medical School) ;
  • Li, Chunhao (Brain Tumor Research Laboratory and Department of Neurosurgery Chonnam National University Research Institute of Medical Sciences, Chonnam National University Hwasun Hopital and Medical School) ;
  • Ryu, Hyang-Hwa (Brain Tumor Research Laboratory and Department of Neurosurgery Chonnam National University Research Institute of Medical Sciences, Chonnam National University Hwasun Hopital and Medical School) ;
  • Lim, Sa-Hoe (Brain Tumor Research Laboratory and Department of Neurosurgery Chonnam National University Research Institute of Medical Sciences, Chonnam National University Hwasun Hopital and Medical School) ;
  • Jang, Woo-Youl (Brain Tumor Research Laboratory and Department of Neurosurgery Chonnam National University Research Institute of Medical Sciences, Chonnam National University Hwasun Hopital and Medical School) ;
  • Jung, Shin (Brain Tumor Research Laboratory and Department of Neurosurgery Chonnam National University Research Institute of Medical Sciences, Chonnam National University Hwasun Hopital and Medical School)
  • Received : 2015.09.23
  • Accepted : 2016.01.21
  • Published : 2016.03.01
Download PDF
⟨ Previous Next ⟩

Abstract

Objective : Protein disulfide isomerase (PDI) acts as a chaperone on the cell surface, and it has been reported that PDI is associated with the tumor cell migration and invasion. The aims of this study are to investigate the anti-migration effect of bacitracin, which is an inhibitor of PDI, and the associated factor in this process. Methods : U87-MG glioma cells were treated with bacitracin in 1.25, 2.5, 3.75, and 5.0 mM concentrations. Western blot with caspase-3 was applied to evaluate the cytotoxicity of bacitracin. Adhesion, morphology, migration assays, and organotypic brain-slice culture were performed to evaluate the effect of bacitracin to the tumor cell. Western blot, PCR, and gelatin zymography were performed to investigate the associated factors. Thirty glioma tissues were collected following immunohistochemistry and Western blot. Results : Bacitracin showed a cytotoxicity in 3rd (p<0.05) and 4th (p<0.001) days, in 5.0 Mm concentration. The cell adhesion significantly decreased and the cells became a round shape after treated with bacitracin. The migration ability, the expression of phosphorylated focal adhesion kinase (p-FAK) and matrix metalloproteinase-2 (MMP-2) decreased in a bacitracin dose- and time-dependent manner. The U87-MG cells exhibited low-invasiveness in the 2.5 mM, compared with the untreated in organotypic brain-slice culture. PDI was expressed in the tumor margin, and significantly increased with histological glioma grades (p<0.001). Conclusion : Bacitracin, as a functional inhibitor of PDI, decreased the phosphorylated FAK and the secreted MMP-2, which are the downstream of integrin and play a major role in cell migration and invasion, might become one of the feasible therapeutic strategies for glioblastoma.

Keywords

References

  1. Benham AM : The protein disulfide isomerase family : key players in health and disease. Antioxid Redox Signal 16 : 781-789, 2012 https://doi.org/10.1089/ars.2011.4439
  2. Deep G, Kumar R, Jain AK, Agarwal C, Agarwal R : Silibinin inhibits fibronectin induced motility, invasiveness and survival in human prostate carcinoma PC3 cells via targeting integrin signaling. Mutat Res 768 : 35-46, 2014 https://doi.org/10.1016/j.mrfmmm.2014.05.002
  3. Di Santo N, Ehrisman J : Research perspective : potential role of nitazoxanide in ovarian cancer treatment. Old drug, new purpose? Cancers (Basel) 5 : 1163-1176, 2013 https://doi.org/10.3390/cancers5031163
  4. Dickerhof N, Kleffmann T, Jack R, McCormick S : Bacitracin inhibits the reductive activity of protein disulfide isomerase by disulfide bond formation with free cysteines in the substrate-binding domain. FEBS J 278 : 2034-2043, 2011 https://doi.org/10.1111/j.1742-4658.2011.08119.x
  5. Essex DW : The role of thiols and disulfides in platelet function. Antioxid Redox Signal 6 : 736-746, 2004 https://doi.org/10.1089/1523086041361622
  6. Essex DW, Chen K, Swiatkowska M : Localization of protein disulfide isomerase to the external surface of the platelet plasma membrane. Blood 86 : 2168-2173, 1995
  7. Goplen D, Wang J, Enger PO, Tysnes BB, Terzis AJ, Laerum OD, et al. : Protein disulfide isomerase expression is related to the invasive properties of malignant glioma. Cancer Res 66 : 9895-9902, 2006 https://doi.org/10.1158/0008-5472.CAN-05-4589
  8. Hatahet F, Ruddock LW : Substrate recognition by the protein disulfide isomerases. FEBS J 274 : 5223-5234, 2007 https://doi.org/10.1111/j.1742-4658.2007.06058.x
  9. Huang G, Ho B, Conroy J, Liu S, Qiang H, Golubovskaya V : The microarray gene profiling analysis of glioblastoma cancer cells reveals genes affected by FAK inhibitor Y15 and combination of Y15 and temozolomide. Anticancer Agents Med Chem 14 : 9-17, 2014 https://doi.org/10.2174/18715206113139990141
  10. Janouskova H, Maglott A, Leger DY, Bossert C, Noulet F, Guerin E, et al. : Integrin ${\alpha}5{\beta}1$ plays a critical role in resistance to temozolomide by interfering with the p53 pathway in high-grade glioma. Cancer Res 72 : 3463-3470, 2012 https://doi.org/10.1158/0008-5472.CAN-11-4199
  11. Jiang XM, Fitzgerald M, Grant CM, Hogg PJ : Redox control of exofacial protein thiols/disulfides by protein disulfide isomerase. J Biol Chem 274 : 2416-2423, 1999 https://doi.org/10.1074/jbc.274.4.2416
  12. Johnson GG, White MC, Grimaldi M : Stressed to death : targeting endoplasmic reticulum stress response induced apoptosis in gliomas. Curr Pharm Des 17 : 284-292, 2011 https://doi.org/10.2174/138161211795049660
  13. Johnson GG, White MC, Wu JH, Vallejo M, Grimaldi M : The deadly connection between endoplasmic reticulum, Ca2+, protein synthesis, and the endoplasmic reticulum stress response in malignant glioma cells. Neuro Oncol 16 : 1086-1099, 2014 https://doi.org/10.1093/neuonc/nou012
  14. Jung S, Kim HW, Lee JH, Kang SS, Rhu HH, Jeong YI, et al. : Brain tumor invasion model system using organotypic brain-slice culture as an alternative to in vivo model. J Cancer Res Clin Oncol 128 : 469-476, 2002 https://doi.org/10.1007/s00432-002-0366-x
  15. Jung TY, Jung S, Ryu HH, Jeong YI, Jin YH, Jin SG, et al. : Role of galectin-1 in migration and invasion of human glioblastoma multiforme cell lines. J Neurosurg 109 : 273-284, 2008 https://doi.org/10.3171/JNS/2008/109/8/0273
  16. Karala AR, Ruddock LW : Bacitracin is not a specific inhibitor of protein disulfide isomerase. FEBS J 277 : 2454-2462, 2010 https://doi.org/10.1111/j.1742-4658.2010.07660.x
  17. Kim CS, Jung S, Jung TY, Jang WY, Sun HS, Ryu HH : Characterization of invading glioma cells using molecular analysis of leading-edge tissue. J Korean Neurosurg Soc 50 : 157-165, 2011 https://doi.org/10.3340/jkns.2011.50.3.157
  18. Lahav J, Gofer-Dadosh N, Luboshitz J, Hess O, Shaklai M : Protein disulfide isomerase mediates integrin-dependent adhesion. FEBS Lett 475 : 89-92, 2000 https://doi.org/10.1016/S0014-5793(00)01630-6
  19. Lahav J, Wijnen EM, Hess O, Hamaia SW, Griffiths D, Makris M, et al. : Enzymatically catalyzed disulfide exchange is required for platelet adhesion to collagen via integrin alpha2beta1. Blood 102 : 2085-2092, 2003 https://doi.org/10.1182/blood-2002-06-1646
  20. Li X, Ishihara S, Yasuda M, Nishioka T, Mizutani T, Ishikawa M, et al. : Lung cancer cells that survive ionizing radiation show increased integrin ${\alpha}2{\beta}1$- and EGFR-dependent invasiveness. PLoS One 8 : e70905, 2013 https://doi.org/10.1371/journal.pone.0070905
  21. Liddington RC, Ginsberg MH : Integrin activation takes shape. J Cell Biol 158 : 833-839, 2002 https://doi.org/10.1083/jcb.200206011
  22. Lovat PE, Corazzari M, Armstrong JL, Martin S, Pagliarini V, Hill D, et al. : Increasing melanoma cell death using inhibitors of protein disulfide isomerases to abrogate survival responses to endoplasmic reticulum stress. Cancer Res 68 : 5363-5369, 2008 https://doi.org/10.1158/0008-5472.CAN-08-0035
  23. Ma J, Cui W, He SM, Duan YH, Heng LJ, Wang L, et al. : Human U87 astrocytoma cell invasion induced by interaction of ${\beta}ig-h3$ with integrin ${\alpha}5{\beta}1$ involves calpain-2. PLoS One 7 : e37297, 2012 https://doi.org/10.1371/journal.pone.0037297
  24. Mandel R, Ryser HJ, Ghani F, Wu M, Peak D : Inhibition of a reductive function of the plasma membrane by bacitracin and antibodies against protein disulfide-isomerase. Proc Natl Acad Sci U S A 90 : 4112-4116, 1993 https://doi.org/10.1073/pnas.90.9.4112
  25. Ryu HH, Jung S, Jung TY, Moon KS, Kim IY, Jeong YI, et al. : Role of metallothionein 1E in the migration and invasion of human glioma cell lines. Int J Oncol 41 : 1305-1313, 2012 https://doi.org/10.3892/ijo.2012.1570
  26. Shi Q, Hjelmeland AB, Keir ST, Song L, Wickman S, Jackson D, et al. : A novel low-molecular weight inhibitor of focal adhesion kinase, TAE226, inhibits glioma growth. Mol Carcinog 46 : 488-496, 2007 https://doi.org/10.1002/mc.20297
  27. Shin BK, Wang H, Yim AM, Le Naour F, Brichory F, Jang JH, et al. : Global profiling of the cell surface proteome of cancer cells uncovers an abundance of proteins with chaperone function. J Biol Chem 278 : 7607-7616, 2003 https://doi.org/10.1074/jbc.M210455200
  28. Sun S, Lee D, Ho AS, Pu JK, Zhang XQ, Lee NP, et al. : Inhibition of prolyl 4-hydroxylase, beta polypeptide (P4HB) attenuates temozolomide resistance in malignant glioma via the endoplasmic reticulum stress response (ERSR) pathways. Neuro Oncol 15 : 562-577, 2013 https://doi.org/10.1093/neuonc/not005
  29. Terada K, Manchikalapudi P, Noiva R, Jauregui HO, Stockert RJ, Schilsky ML : Secretion, surface localization, turnover, and steady state expression of protein disulfide isomerase in rat hepatocytes. J Biol Chem 270 : 20410-20416, 1995 https://doi.org/10.1074/jbc.270.35.20410
  30. Tian F, Zhou X, Wikstrom J, Karlsson H, Sjoland H, Gan LM, et al. : Protein disulfide isomerase increases in myocardial endothelial cells in mice exposed to chronic hypoxia : a stimulatory role in angiogenesis. Am J Physiol Heart Circ Physiol 297 : H1078-H1086, 2009 https://doi.org/10.1152/ajpheart.00937.2008
  31. Tysnes BB, Mahesparan R : Biological mechanisms of glioma invasion and potential therapeutic targets. J Neurooncol 53 : 129-147, 2001 https://doi.org/10.1023/A:1012249216117
  32. Wang EJ, Snyder RD, Fielden MR, Smith RJ, Gu YZ : Validation of putative genomic biomarkers of nephrotoxicity in rats. Toxicology 246 : 91-100, 2008 https://doi.org/10.1016/j.tox.2007.12.031
  33. Weston BS, Wahab NA, Roberts T, Mason RM : Bacitracin inhibits fibronectin matrix assembly by mesangial cells in high glucose. Kidney Int 60 : 1756-1764, 2001 https://doi.org/10.1046/j.1523-1755.2001.00991.x
  34. Wild-Bode C, Weller M, Rimner A, Dichgans J, Wick W : Sublethal irradiation promotes migration and invasiveness of glioma cells : implications for radiotherapy of human glioblastoma. Cancer Res 61 : 2744-2750, 2001
  35. Xu S, Sankar S, Neamati N : Protein disulfide isomerase : a promising target for cancer therapy. Drug Discov Today 19 : 222-240, 2014 https://doi.org/10.1016/j.drudis.2013.10.017
  36. Yu SJ, Yoon JH, Yang JI, Cho EJ, Kwak MS, Jang ES, et al. : Enhancement of hexokinase II inhibitor-induced apoptosis in hepatocellular carcinoma cells via augmenting ER stress and anti-angiogenesis by protein disulfide isomerase inhibition. J Bioenerg Biomembr 44 : 101-115, 2012 https://doi.org/10.1007/s10863-012-9416-5
  37. Zhang K, Ding W, Sun W, Sun XJ, Xie YZ, Zhao CQ, et al. : Beta1 integrin inhibits apoptosis induced by cyclic stretch in annulus fibrosus cells via ERK1/2 MAPK pathway. Apoptosis, 2015 [Epub ahead of print]

Cited by

  1. Enhancement of radiosensitivity by inhibition of c-Jun N-terminal kinase activity in a Lewis lung carcinoma-bearing subcutaneous tumor mouse model vol.36, pp.6, 2016, https://doi.org/10.3892/or.2016.5204
  2. Upregulated expression of MMP-9 in gingival epithelial cells induced by prolonged stimulation with arecoline vol.14, pp.1, 2016, https://doi.org/10.3892/ol.2017.6194
  3. Ganoderma lucidum polysaccharide inhibits prostate cancer cell migration via the protein arginine methyltransferase 6 signaling pathway vol.17, pp.1, 2016, https://doi.org/10.3892/mmr.2017.7904
  4. P4HB and PDIA3 are associated with tumor progression and therapeutic outcome of diffuse gliomas vol.39, pp.2, 2016, https://doi.org/10.3892/or.2017.6134
  5. Design, Synthesis, and Biological Evaluation of Novel Allosteric Protein Disulfide Isomerase Inhibitors vol.62, pp.7, 2016, https://doi.org/10.1021/acs.jmedchem.8b01951
  6. Curcumin inhibits the expression of proinflammatory mediators and MMP-9 in gingival epithelial cells stimulated for a prolonged period with lipopolysaccharides derived from Porphyromonas gingivalis vol.108, pp.1, 2020, https://doi.org/10.1007/s10266-019-00432-8
  7. Protein disulfide isomerases are promising targets for predicting the survival and tumor progression in glioma patients vol.12, pp.3, 2016, https://doi.org/10.18632/aging.102748
  8. DNA hypermethylation of sirtuin 1 ( SIRT1 ) caused by betel quid chewing—a possible predictive biomarker for malignant transformation vol.12, pp.None, 2016, https://doi.org/10.1186/s13148-019-0806-y
  9. ceRNA Network Analysis Shows That lncRNA CRNDE Promotes Progression of Glioblastoma Through Sponge mir-9-5p vol.12, pp.None, 2021, https://doi.org/10.3389/fgene.2021.617350
  10. Zafirlukast is a broad‐spectrum thiol isomerase inhibitor that inhibits thrombosis without altering bleeding times vol.178, pp.3, 2016, https://doi.org/10.1111/bph.15291
  11. Differential expression of protein disulfide isomerase (PDI) in regulating endometrial receptivity in humans vol.21, pp.2, 2021, https://doi.org/10.1016/j.repbio.2021.100498
  12. Bacitracin is a non-competitive inhibitor of porcine M1 family neutral and glutamyl aminopeptidases vol.35, pp.17, 2016, https://doi.org/10.1080/14786419.2019.1678611