Abstract
The multifunctional protein - tissue inhibitor of metalloproteinases-1 (TIMP-1) - has been associated with a poor prognosis in several types of cancers including glioblastomas. In addition, TIMP-1 has been associated with decreased response to chemotherapy, and especially the efficacy of the family of topoisomerase (TOP) inhibitors has been related to TIMP-1. As a second line treatment of glioblastomas, the vascular endothelial growth factor (VEGF) antibody bevacizumab is administered in combination with the TOP1 inhibitor irinotecan and glioblastoma cell levels of TIMP-1 could therefore potentially influence the efficacy of such treatment. In the present study, we aimed to investigate whether a high TIMP-1 expression in glioblastoma cell lines would affect the sensitivity to TOP inhibitors, and whether TIMP-1 overexpressing cells would have alterered growth and invasion. We established TIMP-1 overexpressing subclones from two human glioblastoma cell lines. TIMP-1 overexpressing U87MG cells were significantly more resistant than low TIMP-1 expressing clones and parental cells when exposed to SN-38 (TOP1 inhibitor) or epirubicin (TOP2 inhibitor). No significant differences were observed for the TIMP-1 transfected A172 cells. Implantation of both U87MG and A172 spheroids into organotypic brain slice cultures revealed a reduced growth of TIMP-1 overexpressing U87MG spheroids, however, no significant differences in invasion were observed. The present study suggests that TIMP-1 overexpression reduces the effect of TOP inhibitors in glioblastoma. TIMP-1 also appeared to reduce spheroid growth, but did not influence invasion. Whether TIMP-1 plays a role in irinotecan resistance and has a predictive potential in glioblastoma patients remains to be elucidated.
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References
Louis DNW, Cavenee WK (2007) WHO classification of tumors of the central nervous system, 4 edn. International Agency for Research on Cancer (IARC), Lyon
Agnihotri S, Gajadhar AS, Ternamian C, Gorlia T, Diefes KL, Mischel PS, Kelly J, McGown G, Thorncroft M, Carlson BL, Sarkaria JN, Margison GP, Aldape K, Hawkins C, Hegi M, Guha A (2012) Alkylpurine-DNA-N-glycosylase confers resistance to temozolomide in xenograft models of glioblastoma multiforme and is associated with poor survival in patients. J Clin Invest 122(1):253–266. https://doi.org/10.1172/JCI59334
Bao S, Wu Q, McLendon RE, Hao Y, Shi Q, Hjelmeland AB, Dewhirst MW, Bigner DD, Rich JN (2006) Glioma stem cells promote radioresistance by preferential activation of the DNA damage response. Nature 444(7120):756–760. https://doi.org/10.1038/nature05236
Dean M (2009) ABC transporters, drug resistance, and cancer stem cells. J Mammary Gland Biol Neoplasia 14(1):3–9. https://doi.org/10.1007/s10911-009-9109-9
Dean M, Fojo T, Bates S (2005) Tumour stem cells and drug resistance. Nat Rev Cancer 5(4):275–284. https://doi.org/10.1038/nrc1590
Esteller M, Garcia-Foncillas J, Andion E, Goodman SN, Hidalgo OF, Vanaclocha V, Baylin SB, Herman JG (2000) Inactivation of the DNA-repair gene MGMT and the clinical response of gliomas to alkylating agents. N Engl J Med 343(19):1350–1354. https://doi.org/10.1056/nejm200011093431901
Nakai E, Park K, Yawata T, Chihara T, Kumazawa A, Nakabayashi H, Shimizu K (2009) Enhanced MDR1 expression and chemoresistance of cancer stem cells derived from glioblastoma. Cancer Investig 27(9):901–908. https://doi.org/10.3109/07357900801946679
Salmaggi A, Boiardi A, Gelati M, Russo A, Calatozzolo C, Ciusani E, Sciacca FL, Ottolina A, Parati EA, La Porta C, Alessandri G, Marras C, Croci D, De Rossi M (2006) Glioblastoma-derived tumorospheres identify a population of tumor stem-like cells with angiogenic potential and enhanced multidrug resistance phenotype. Glia 54(8):850–860. https://doi.org/10.1002/glia.20414
Chirco R, Liu XW, Jung KK, Kim HR (2006) Novel functions of TIMPs in cell signaling. Cancer Metastasis Rev 25(1):99–113. https://doi.org/10.1007/s10555-006-7893-x
Egeblad M, Werb Z (2002) New functions for the matrix metalloproteinases in cancer progression. Nat Rev Cancer 2(3):161–174. https://doi.org/10.1038/nrc745
Gomez DE, Alonso DF, Yoshiji H, Thorgeirsson UP (1997) Tissue inhibitors of metalloproteinases: structure, regulation and biological functions. Eur J Cell Biol 74(2):111–122
Jiang Y, Goldberg ID, Shi YE (2002) Complex roles of tissue inhibitors of metalloproteinases in cancer. Oncogene 21(14):2245–2252. https://doi.org/10.1038/sj.onc.1205291
Bloomston M, Shafii A, Zervos E, Rosemurgy AS (2005) TIMP-1 antisense gene transfection attenuates the invasive potential of pancreatic cancer cells in vitro and inhibits tumor growth in vivo. Am J Surg 189(6):675–679. https://doi.org/10.1016/j.amjsurg.2005.03.008
Guedez L, McMarlin AJ, Kingma DW, Bennett TA, Stetler-Stevenson M, Stetler-Stevenson WG (2001) Tissue inhibitor of metalloproteinase-1 alters the tumorigenicity of Burkitt's lymphoma via divergent effects on tumor growth and angiogenesis. Am J Pathol 158(4):1207–1215. https://doi.org/10.1016/s0002-9440(10)64070-9
Jung KK, Liu XW, Chirco R, Fridman R, Kim HR (2006) Identification of CD63 as a tissue inhibitor of metalloproteinase-1 interacting cell surface protein. EMBO J 25(17):3934–3942. https://doi.org/10.1038/sj.emboj.7601281
Lambert E, Dasse E, Haye B, Petitfrere E (2004) TIMPs as multifacial proteins. Crit Rev Oncol Hematol 49(3):187–198. https://doi.org/10.1016/j.critrevonc.2003.09.008
Stetler-Stevenson WG (2008) Tissue inhibitors of metalloproteinases in cell signaling: metalloproteinase-independent biological activities. Sci Signal 1(27):re6. https://doi.org/10.1126/scisignal.127re6
Yoshiji H, Harris SR, Raso E, Gomez DE, Lindsay CK, Shibuya M, Sinha CC, Thorgeirsson UP (1998) Mammary carcinoma cells over-expressing tissue inhibitor of metalloproteinases-1 show enhanced vascular endothelial growth factor expression. International journal of cancer Journal international du cancer 75(1):81–87
Liu XW, Bernardo MM, Fridman R, Kim HR (2003) Tissue inhibitor of metalloproteinase-1 protects human breast epithelial cells against intrinsic apoptotic cell death via the focal adhesion kinase/phosphatidylinositol 3-kinase and MAPK signaling pathway. J Biol Chem 278(41):40364–40372. https://doi.org/10.1074/jbc.M302999200
Birgisson H, Nielsen HJ, Christensen IJ, Glimelius B, Brunner N (2010) Preoperative plasma TIMP-1 is an independent prognostic indicator in patients with primary colorectal cancer: a prospective validation study. Eur J Cancer 46(18):3323–3331. https://doi.org/10.1016/j.ejca.2010.06.009
Holten-Andersen MN, Stephens RW, Nielsen HJ, Murphy G, Christensen IJ, Stetler-Stevenson W, Brunner N (2000) High preoperative plasma tissue inhibitor of metalloproteinase-1 levels are associated with short survival of patients with colorectal cancer. Clin Cancer Res : an official journal of the American Association for Cancer Research 6(11):4292–4299
Kuvaja P, Wurtz SO, Talvensaari-Mattila A, Brunner N, Paakko P, Turpeenniemi-Hujanen T (2007) High serum TIMP-1 correlates with poor prognosis in breast carcinoma - a validation study. Cancer Biomark : section A of Disease markers 3(6):293–300
Pesta M, Kulda V, Kucera R, Pesek M, Vrzalova J, Liska V, Pecen L, Treska V, Safranek J, Prazakova M, Vycital O, Bruha J, Holubec L, Topolcan O (2011) Prognostic significance of TIMP-1 in non-small cell lung cancer. Anticancer Res 31(11):4031–4038
Rauvala M, Puistola U, Turpeenniemi-Hujanen T (2005) Gelatinases and their tissue inhibitors in ovarian tumors; TIMP-1 is a predictive as well as a prognostic factor. Gynecol Oncol 99(3):656–663. https://doi.org/10.1016/j.ygyno.2005.07.009
Schrohl AS, Holten-Andersen MN, Peters HA, Look MP, Meijer-van Gelder ME, Klijn JG, Brunner N, Foekens JA (2004) Tumor tissue levels of tissue inhibitor of metalloproteinase-1 as a prognostic marker in primary breast cancer. Clin Cancer Res : an official journal of the American Association for Cancer Research 10(7):2289–2298
Wang CS, Wu TL, Tsao KC, Sun CF (2006) Serum TIMP-1 in gastric cancer patients: a potential prognostic biomarker. Ann Clin Lab Sci 36(1):23–30
Aaberg-Jessen C, Christensen K, Offenberg H, Bartels A, Dreehsen T, Hansen S, Schroder HD, Brunner N, Kristensen BW (2009) Low expression of tissue inhibitor of metalloproteinases-1 (TIMP-1) in glioblastoma predicts longer patient survival. J Neuro-Oncol 95(1):117–128. https://doi.org/10.1007/s11060-009-9910-8
Ejlertsen B, Jensen MB, Nielsen KV, Balslev E, Rasmussen BB, Willemoe GL, Hertel PB, Knoop AS, Mouridsen HT, Brunner N (2010) HER2, TOP2A, and TIMP-1 and responsiveness to adjuvant anthracycline-containing chemotherapy in high-risk breast cancer patients. J Clin Oncol : official journal of the American Society of Clinical Oncology 28(6):984–990. https://doi.org/10.1200/jco.2009.24.1166
Frederiksen C, Qvortrup C, Christensen IJ, Glimelius B, Berglund A, Jensen BV, Nielsen SE, Keldsen N, Nielsen HJ, Brunner N, Pfeiffer P (2011) Plasma TIMP-1 levels and treatment outcome in patients treated with XELOX for metastatic colorectal cancer. Ann Oncol : official journal of the European Society for Medical Oncology / ESMO 22(2):369–375. https://doi.org/10.1093/annonc/mdq354
Klintman M, Ornbjerg Wurtz S, Christensen IJ, Braemer Hertel P, Ferno M, Malmberg M, Mouridsen H, Cold F, Schrohl AS, Foekens JA, Malmstrom P, Brunner N (2010) Association between tumor tissue TIMP-1 levels and objective response to first-line chemotherapy in metastatic breast cancer. Breast Cancer Res Treat 121(2):365–371. https://doi.org/10.1007/s10549-009-0483-1
Schrohl AS, Meijer-van Gelder ME, Holten-Andersen MN, Christensen IJ, Look MP, Mouridsen HT, Brunner N, Foekens JA (2006) Primary tumor levels of tissue inhibitor of metalloproteinases-1 are predictive of resistance to chemotherapy in patients with metastatic breast cancer. Clin Cancer Res : an official journal of the American Association for Cancer Research 12(23):7054–7058. https://doi.org/10.1158/1078-0432.ccr-06-0950
Sorensen NM, Bystrom P, Christensen IJ, Berglund A, Nielsen HJ, Brunner N, Glimelius B (2007) TIMP-1 is significantly associated with objective response and survival in metastatic colorectal cancer patients receiving combination of irinotecan, 5-fluorouracil, and folinic acid. Clin Cancer Res : an official journal of the American Association for Cancer Research 13(14):4117–4122. https://doi.org/10.1158/1078-0432.ccr-07-0186
Gruppe DNO (2012) Retningslinier for behandling af intrakranielle gliomer hos voksne. http://www.dnog.dk/assets/files/Retningslinier%20PDF/DNOG%20gliom%20retningslinje%202012.pdf
Champoux JJ (2000) Structure-based analysis of the effects of camptothecin on the activities of human topoisomerase I. Ann N Y Acad Sci 922:56–64
Hsiang YH, Liu LF (1988) Identification of mammalian DNA topoisomerase I as an intracellular target of the anticancer drug camptothecin. Cancer Res 48(7):1722–1726
Deng X, Fogh L, Lademann U, Jensen V, Stenvang J, Yang H, Brunner N, Schrohl AS (2013) TIMP-1 overexpression does not affect sensitivity to HER2-targeting drugs in the HER2-gene-amplified SK-BR-3 human breast cancer cell line. Tumour Biol : the journal of the International Society for Oncodevelopmental Biology and Medicine 34(2):1161–1170. https://doi.org/10.1007/s13277-013-0659-5
Holten-Andersen MN, Christensen IJ, Nielsen HJ, Lilja H, Murphy G, Jensen V, Brunner N, Piironen T (2002) Measurement of the noncomplexed free fraction of tissue inhibitor of metalloproteinases 1 in plasma by immunoassay. Clin Chem 48(8):1305–1313
Holten-Andersen MN, Murphy G, Nielsen HJ, Pedersen AN, Christensen IJ, Hoyer-Hansen G, Brunner N, Stephens RW (1999) Quantitation of TIMP-1 in plasma of healthy blood donors and patients with advanced cancer. Br J Cancer 80(3–4):495–503. https://doi.org/10.1038/sj.bjc.6690384
Aaberg-Jessen C, Norregaard A, Christensen K, Pedersen CB, Andersen C, Kristensen BW (2013) Invasion of primary glioma- and cell line-derived spheroids implanted into corticostriatal slice cultures. Int J Clin Exp Pathol 6(4):546–560
Hekmat O, Munk S, Fogh L, Yadav R, Francavilla C, Horn H, Wurtz SO, Schrohl AS, Damsgaard B, Romer MU, Belling KC, Jensen NF, Gromova I, Bekker-Jensen DB, Moreira JM, Jensen LJ, Gupta R, Lademann U, Brunner N, Olsen JV, Stenvang J (2013) TIMP-1 increases expression and phosphorylation of proteins associated with drug resistance in breast cancer cells. J Proteome Res. https://doi.org/10.1021/pr400457u
Fu ZY, Lv JH, Ma CY, Yang DP, Wang T (2011) Tissue inhibitor of metalloproteinase-1 decreased chemosensitivity of MDA-435 breast cancer cells to chemotherapeutic drugs through the PI3K/AKT/NF-small ka, CyrillicB pathway. Biomed Pharmacother = Biomedecine & pharmacotherapie 65(3):163–167. https://doi.org/10.1016/j.biopha.2011.02.004
Wang T, Lv JH, Zhang XF, Li CJ, Han X, Sun YJ (2010) Tissue inhibitor of metalloproteinase-1 protects MCF-7 breast cancer cells from paclitaxel-induced apoptosis by decreasing the stability of cyclin B1. Int J Cancer: Journal international du cancer 126(2):362–370. https://doi.org/10.1002/ijc.24753
Willemoe GL, Hertel PB, Bartels A, Jensen MB, Balslev E, Rasmussen BB, Mouridsen H, Ejlertsen B, Brunner N (2009) Lack of TIMP-1 tumour cell immunoreactivity predicts effect of adjuvant anthracycline-based chemotherapy in patients (n=647) with primary breast cancer. A Danish breast cancer cooperative group study. Eur J Cancer 45(14):2528–2536. https://doi.org/10.1016/j.ejca.2009.05.029
Galli R, Binda E, Orfanelli U, Cipelletti B, Gritti A, De Vitis S, Fiocco R, Foroni C, Dimeco F, Vescovi A (2004) Isolation and characterization of tumorigenic, stem-like neural precursors from human glioblastoma. Cancer Res 64(19):7011–7021. https://doi.org/10.1158/0008-5472.can-04-1364
Lee J, Kotliarova S, Kotliarov Y, Li A, Su Q, Donin NM, Pastorino S, Purow BW, Christopher N, Zhang W, Park JK, Fine HA (2006) Tumor stem cells derived from glioblastomas cultured in bFGF and EGF more closely mirror the phenotype and genotype of primary tumors than do serum-cultured cell lines. Cancer Cell 9(5):391–403. https://doi.org/10.1016/j.ccr.2006.03.030
Yu SC, ** YF, Yi L, Zhou ZH, Chen JH, Yao XH, Gao L, Wang JM, Bian XW (2008) Isolation and characterization of cancer stem cells from a human glioblastoma cell line U87. Cancer Lett 265(1):124–134. https://doi.org/10.1016/j.canlet.2008.02.010
Nakano A, Tani E, Miyazaki K, Yamamoto Y, Furuyama J (1995) Matrix metalloproteinases and tissue inhibitors of metalloproteinases in human gliomas. J Neurosurg 83(2):298–307. https://doi.org/10.3171/jns.1995.83.2.0298
Matsuzawa K, Fukuyama K, Hubbard SL, Dirks PB, Rutka JT (1996) Transfection of an invasive human astrocytoma cell line with a TIMP-1 cDNA: modulation of astrocytoma invasive potential. J Neuropathol Exp Neurol 55(1):88–96
Bigelow RL, Williams BJ, Carroll JL, Daves LK, Cardelli JA (2009) TIMP-1 overexpression promotes tumorigenesis of MDA-MB-231 breast cancer cells and alters expression of a subset of cancer promoting genes in vivo distinct from those observed in vitro. Breast Cancer Res Treat 117(1):31–44. https://doi.org/10.1007/s10549-008-0170-7
Acknowledgements
We would like to thank technicians Helle Wohlleben and Tanja Dreehsen Højgaard for assistance with the immunohistochemistry and Vibeke Jensen for assistance with ELISA.
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The work was supported by The Danish Cancer Society, Danish Cancer Research Foundation and Fonden til Laegevidenskabens Fremme.
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Aaberg-Jessen, C., Fogh, L., Sørensen, M.D. et al. Overexpression of TIMP-1 and Sensitivity to Topoisomerase Inhibitors in Glioblastoma Cell Lines. Pathol. Oncol. Res. 25, 59–69 (2019). https://doi.org/10.1007/s12253-017-0312-5
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DOI: https://doi.org/10.1007/s12253-017-0312-5