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11.04.2017 | Original Paper

Dual treatment with shikonin and temozolomide reduces glioblastoma tumor growth, migration and glial-to-mesenchymal transition

verfasst von: Diana Matias, Joana Balça-Silva, Luiz Gustavo Dubois, Bruno Pontes, Valéria Pereira Ferrer, Luciane Rosário, Anália do Carmo, Juliana Echevarria-Lima, Ana Bela Sarmento-Ribeiro, Maria Celeste Lopes, Vivaldo Moura-Neto

Erschienen in: Cellular Oncology | Ausgabe 3/2017

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Abstract

Purpose

Glioblastomas (GBM) comprise 17% of all primary brain tumors. These tumors are extremely aggressive due to their infiltrative capacity and chemoresistance, with glial-to-mesenchymal transition (GMT) proteins playing a prominent role in tumor invasion. One compound that has recently been used to reduce the expression of these proteins is shikonin (SHK), a naphthoquinone with anti-tumor properties. Temozolomide (TMZ), the most commonly used chemotherapeutic agent in GBM treatment, has so far not been studied in combination with SHK. Here, we investigated the combined effects of these two drugs on the proliferation and motility of GBM-derived cells.

Methods

The cytotoxic and proliferative effects of SHK and TMZ on human GBM-derived cells were tested using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), Ki67 staining and BrdU incorporation assays. The migration capacities of these cells were evaluated using a scratch wound assay. The expression levels of β3 integrin, metalloproteinases (MMPs) and GMT-associated proteins were determined by Western blotting and immunocytochemistry.

Results

We found that GBM-derived cells treated with a combination of SHK and TMZ showed decreases in their proliferation and migration capacities. These decreases were followed by the suppression of GMT through a reduction of β3 integrin, MMP-2, MMP-9, Slug and vimentin expression via inactivation of PI3K/AKT signaling.

Conclusion

From our results we conclude that dual treatment with SHK and TMZ may constitute a powerful new tool for GBM treatment by reducing therapy resistance and tumor recurrence.

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F.B. Furnari, T. Fenton, R.M. Bachoo, A. Mukasa, J.M. Stommel, A. Stegh, W.C. Hahn, K.L. Ligon, D.N. Louis, C. Brennan, L. Chin, R.A. DePinho, W.K. Cavenee, Malignant astrocytic glioma: Genetics, biology, and paths to treatment. Genes. Dev. 21, 2683–2710 (2007). doi:10.1101/gad.1596707 CrossRefPubMed

F.R. Lima, S.A. Kahn, R.C. Soletti, D. Biasoli, T. Alves, A.C. da Fonseca, C. Garcia, L. Romao, J. Brito, R. Holanda-Afonso, J. Faria, H. Borges, V. Moura-Neto, Glioblastoma: Therapeutic challenges, what lies ahead. Biochim. Biophys. Acta. 1826, 338–349 (2012). doi:10.1016/j.bbcan.2012.05.004 PubMed

R. Stupp, M. E. Hegi, W. P. Mason, M. J. van den Bent, M. J. Taphoorn, R. C. Janzer, S. K. Ludwin, A. Allgeier, B. Fisher, K. Belanger, P. Hau, A. A. Brandes, J. Gijtenbeek, C. Marosi, C. J. Vecht, K. Mokhtari, P. Wesseling, S. Villa, E. Eisenhauer, T. Gorlia, M. Weller, D. Lacombe, J. G. Cairncross, R. O. Mirimanoff, European Organisation for Research, Treatment of Cancer Brain Tumour and Radiation Oncology Groups, National Cancer Institute of Canada Clinical Trials Group. Effects of radiotherapy with concomitant and adjuvant temozolomide versus radiotherapy alone on survival in glioblastoma in a randomised phase III study: 5-year analysis of the EORTC-NCIC trial. Lancet. Oncol. 10, 459–466 (2009) doi: 10.1016/S1470–2045(09)70025–7

S.A. Kahn, D. Biasoli, C. Garcia, L.H. Geraldo, B. Pontes, M. Sobrinho, A.C. Frauches, L. Romao, R.C. Soletti, S. Assuncao Fdos, F. Tovar-Moll, J.M. de Souza, F.R. Lima, G. Anderluh, V. Moura-Neto, Equinatoxin II potentiates temozolomide- and etoposide-induced glioblastoma cell death. Current. Top. Med. Chem. 12, 2082–2093 (2012)CrossRef

V. Moura-Neto, L. Campanati, D. Matias, C.M. Pereira, C. Freitas, J. M. Coelho-Aguiar, T.C.L.S. Spohr, A.L. Tavares-Gomes, D. Pinheiro-Aguiar, S.A. Kahn, J. Balça-Silva, B. Pontes, I. Porto-Carreiro, J. Faria, R.A.P. Martins, S. Lima-Costa, M. F. Dias-Costa, M.C. Lopes, F.R.S. Lima. Glioblastoma and the special role of adhesion molecules in their invasion, ed. by A. Sedo, R. Mentlein (Springer-Verlag Wien, 2014), p. 293. doi:10.1007/978–3–7091-1431-5

H.J. Anderson, D.S. Galileo, Small-molecule inhibitors of FGFR, integrins and FAK selectively decrease L1CAM-stimulated glioblastoma cell motility and proliferation. Cell. Oncol. 39, 229–242 (2016). doi:10.1007/s13402-016-0267-7

B.N. Smith, N.A. Bhowmick, Role of EMT in metastasis and therapy resistance. J. Clin. Med. 5, pii:E17 (2016). doi:10.3390/jcm5020017

J.S. Desgrosellier, D.A. Cheresh, Integrins in cancer: Biological implications and therapeutic opportunities. Nat. Rev. Cancer 10, 9–22 (2010). doi:10.1038/nrc2748

F.A. Mamuya, M.K. Duncan, aV integrins and TGF-β-induced EMT: A circle of regulation. J. Cell. Mol. Med. 16, 445–455 (2012). doi:10.1111/j.1582-4934.2011.01419.x CrossRefPubMedPubMedCentral

10.

C.Y. Liu, H.H. Lin, M.J. Tang, Y.K. Wang. Vimentin contributes to epithelial-mesenchymal transition cancer cell mechanics by mediating cytoskeletal organization and focal adhesion maturation. Oncotarget 6, 15966–15983 (2015) doi:10.18632/oncotarget.3862

11.

L.J. McCawley, L.M. Matrisian, Matrix metalloproteinases: Multifunctional contributors to tumor progression. Mol. Med. Today 6, 149–156 (2000). doi:10.1016/S1357-4310(00)01686-5

12.

E. Godefroy, A. Moreau-Aubry, E. Diez, B. Dreno, F. Jotereau, Y. Guilloux. alpha v beta3-dependent cross-presentation of matrix metalloproteinase-2 by melanoma cells gives rise to a new tumor antigen. J. Exp. Med. 202, 61–72 (2005) doi: 10.1084/jem.20042138

13.

R.C. Soletti, G.P. de Faria, J. Vernal, H. Terenzi, G. Anderluh, H.L. Borges, V. Moura-Neto, N.H. Gabilan, Potentiation of anticancer-drug cytotoxicity by sea anemone pore-forming proteins in human glioblastoma cells. Anti-Cancer Drugs 19, 517–525 (2008). doi:10.1097/CAD.0b013e3282faa704

14.

B.L. Santos, M.N. Oliveira, P.L. Coelho, B.P. Pitanga, A.B. da Silva, T. Adelita, V.D. Silva, F. Costa Mde. R.S. El-Bacha, M. Tardy, H. Chneiweiss, M.P. Junier, V. Moura-Neto, S.L. Costa. Flavonoids suppress human glioblastoma cell growth by inhibiting cell metabolism, migration, and by regulating extracellular matrix proteins and metalloproteinases expression. Chem. Biol. Interact. 242, 123–138 (2015) doi: 10.1016/j.cbi.2015.07.014

15.

J. Balca-Silva, D. Matias, A. do Carmo, H. Girao, V. Moura-Neto, A.B. Sarmento-Ribeiro, M.C. Lopes. Tamoxifen in combination with temozolomide induce a synergistic inhibition of PKC-pan in GBM cell lines. Biochim. Biophys. Acta 1850, 722–732 (2015) doi:10.1016/j.bbagen.2014.12.022

16.

X. Chen, L. Yang, J.J. Oppenheim, M.Z. Howard, Cellular pharmacology studies of shikonin derivatives. Phytother. Res. 16, 199–209 (2002). doi:10.1002/ptr.1100 CrossRefPubMed

17.

Y. Chen, L. Zheng, J. Liu, Z. Zhou, X. Cao, X. Lv, F. Chen, Shikonin inhibits prostate cancer cells metastasis by reducing matrix metalloproteinase-2/−9 expression via AKT/mTOR and ROS/ERK1/2 pathways. Int. Immunopharmacol. 21, 447–455 (2014). doi:10.1016/j.intimp.2014.05.026 CrossRefPubMed

18.

H. Wang, C. Wu, S. Wan, H. Zhang, S. Zhou, G. Liu, Shikonin attenuates lung cancer cell adhesion to extracellular matrix and metastasis by inhibiting integrin β1 expression and the ERK1/2 signaling pathway. Toxicology 308, 104–112 (2013). doi:10.1016/j.tox.2013.03.015

19.

Q. Zhao, N. Kretschmer, R. Bauer, T. Efferth, Shikonin and its derivatives inhibit the epidermal growth factor receptor signaling and synergistically kill glioblastoma cells in combination with erlotinib. Int. J. Cancer 137, 1446–1456 (2015). doi:10.1002/ijc.29483

20.

F.L. Zhang, P. Wang, Y.H. Liu, L.B. Liu, X.B. Liu, Z. Li, Y.X. Xue, Topoisomerase I inhibitors, shikonin and topotecan, inhibit growth and induce apoptosis of glioma cells and glioma stem cells. PLoS One 8, e81815 (2013). doi:10.1371/journal.pone.0081815

21.

D. Hong, S.Y. Jang, E.H. Jang, B. Jung, I.H. Cho, M.J. Park, S.Y. Jeong, J.H. Kim, Shikonin as an inhibitor of the LPS-induced epithelial-to-mesenchymal transition in human breast cancer cells. Int. J. Mol. Med. 36, 1601–1606 (2015). doi:10.3892/ijmm.2015.2373 PubMed

22.

R. Mahabir, M. Tanino, A. Elmansuri, L. Wang, T. Kimura, T. Itoh, Y. Ohba, H. Nishihara, H. Shirato, M. Tsuda, S. Tanaka, Sustained elevation of snail promotes glial-mesenchymal transition after irradiation in malignant glioma. Neuro-Oncology 16, 671–685 (2014). doi:10.1093/neuonc/not239

23.

J. Faria, L. Romao, S. Martins, T. Alves, F.A. Mendes, G.P. de Faria, R. Hollanda, C. Takiya, L. Chimelli, V. Morandi, J.M. de Souza, J.G. Abreu, V. Moura Neto, Interactive properties of human glioblastoma cells with brain neurons in culture and neuronal modulation of glial laminin organization. Differentiation 74, 562–572 (2006). doi:10.1111/j.1432-0436.2006.00090.x

24.

L.F. Romao, F.A. Mendes, N.M. Feitosa, J.C. Faria, J.M. Coelho-Aguiar, J.M. de Souza, V. Moura-Neto, J.G. Abreu, Connective tissue growth factor (CTGF/CCN2) is negatively regulated during neuron-glioblastoma interaction. PLoS One 8, e55605 (2013). doi:10.1371/journal.pone.0055605

25.

C.C. Liang, A.Y. Park, J.L. Guan, In vitro scratch assay: A convenient and inexpensive method for analysis of cell migration in vitro. Nat. Protoc. 2, 329–333 (2007). doi:10.1038/nprot.2007.30

26.

A. Boudaoud, A. Burian, D. Borowska-Wykret, M. Uyttewaal, R. Wrzalik, D. Kwiatkowska, O. Hamant, FibrilTool, an ImageJ plug-in to quantify fibrillar structures in raw microscopy images. Nat. Protoc. 9, 457–463 (2014). doi:10.1038/nprot.2014.024 CrossRefPubMed

27.

H. Towbin, T. Staehelin, J. Gordon, Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Biotechnology 24, 145–149 (1979)

28.

K.A. McDowell, G.J. Riggins, G.L. Gallia, Targeting the AKT pathway in glioblastoma. Curr. Pharm. Des. 17, 2411–2420 (2011)CrossRefPubMed

29.

N. Fenouille, M. Tichet, M. Dufies, A. Pottier, A. Mogha, J.K. Soo, S. Rocchi, A. Mallavialle, M.D. Galibert, A. Khammari, J.P. Lacour, R. Ballotti, M. Deckert, S. Tartare-Deckert, The epithelial-mesenchymal transition (EMT) regulatory factor SLUG (SNAI2) is a downstream target of SPARC and AKT in promoting melanoma cell invasion. PLoS One 7, e40378 (2012). doi:10.1371/journal.pone.0040378

30.

Y. Iwadate, Epithelial-mesenchymal transition in glioblastoma progression. Oncol. Lett. 11, 1615–1620 (2016). doi:10.3892/ol.2016.4113 PubMedPubMedCentral

31.

W.Y. Cheng, J.J. Kandel, D.J. Yamashiro, P. Canoll, D. Anastassiou, A multi-cancer mesenchymal transition gene expression signature is associated with prolonged time to recurrence in glioblastoma. PLoS One 7, e34705 (2012). doi:10.1371/journal.pone.0034705

32.

E.M. Goellner, B. Grimme, A.R. Brown, Y.C. Lin, X.H. Wang, K.F. Sugrue, L. Mitchell, R.N. Trivedi, J.B. Tang, R.W. Sobol, Overcoming temozolomide resistance in glioblastoma via dual inhibition of NAD+ biosynthesis and base excision repair. Cancer Res. 71, 2308–2317 (2011). doi:10.1158/0008-5472.CAN-10-3213

33.

H. Wu, J. Xie, Q. Pan, B. Wang, D. Hu, X. Hu, Anticancer agent shikonin is an incompetent inducer of cancer drug resistance. PLoS One 8, e52706 (2013). doi:10.1371/journal.pone.0052706

34.

W. Li, J. Liu, K. Jackson, R. Shi, Y. Zhao, Sensitizing the therapeutic efficacy of taxol with shikonin in human breast cancer cells. PLoS One 9, e94079 (2014). doi:10.1371/journal.pone.0094079 CrossRefPubMedPubMedCentral

35.

M.E. Hegi, A.C. Diserens, T. Gorlia, M.F. Hamou, N. de Tribolet, M. Weller, J.M. Kros, J.A. Hainfellner, W. Mason, L. Mariani, J.E. Bromberg, P. Hau, R.O. Mirimanoff, J.G. Cairncross, R.C. Janzer, R. Stupp, MGMT gene silencing and benefit from temozolomide in glioblastoma. N. Engl. J. Med. 352, 997–1003 (2005). doi:10.1056/NEJMoa043331 CrossRefPubMed

36.

C. Kubelt, K. Hattermann, S. Sebens, H.M. Mehdorn, J. Held-Feindt, Epithelial-to-mesenchymal transition in paired human primary and recurrent glioblastomas. Int. J. Oncol. 46, 2515–2525 (2015). doi:10.3892/ijo.2015.2944 PubMed

37.

Y.W. Heng, H.H. Lim, T. Mina, P. Utomo, S. Zhong, C.T. Lim, C.G. Koh, TPPP acts downstream of RhoA-ROCK-LIMK2 to regulate astral microtubule organization and spindle orientation. J. Cell. Sci. 125, 1579–1590 (2012). doi:10.1242/jcs.096818 CrossRefPubMed

38.

M. Silginer, M. Weller, U. Ziegler, P. Roth, Integrin inhibition promotes atypical anoikis in glioma cells. Cell. Death Dis. 5, e1012 (2014). doi:10.1038/cddis.2013.543

39.

M. Rolli, E. Fransvea, J. Pilch, A. Saven, B. Felding-Habermann, Activated integrin alphavbeta3 cooperates with metalloproteinase MMP-9 in regulating migration of metastatic breast cancer cells. Proc. Natl. Acad. Sci. U S A 100, 9482–9487 (2003). doi:10.1073/pnas.1633689100 CrossRefPubMedPubMedCentral

40.

S.S. Lakka, C.S. Gondi, N. Yanamandra, W.C. Olivero, D.H. Dinh, M. Gujrati, J.S. Rao, Inhibition of cathepsin B and MMP-9 gene expression in glioblastoma cell line via RNA interference reduces tumor cell invasion, tumor growth and angiogenesis. Oncogene 23, 4681–4689 (2004). doi:10.1038/sj.onc.1207616

41.

C. Chetty, S.S. Lakka, P. Bhoopathi, J.S. Rao, MMP-2 alters VEGF expression via alphaVbeta3 integrin-mediated PI3K/AKT signaling in A549 lung cancer cells. Int. J. Cancer 127, 1081–1095 (2010). doi:10.1002/ijc.25134

42.

A.V. Fonseca, D. Corbeil, The hematopoietic stem cell polarization and migration: A dynamic link between RhoA signaling pathway, microtubule network and ganglioside-based membrane microdomains. Commun. Integr. Biol. 4, 201–204 (2011). doi:10.4161/cib.4.2.14419 CrossRefPubMedPubMedCentral

43.

U. Jadhav, S. Chigurupati, S.S. Lakka, S. Mohanam, Inhibition of matrix metalloproteinase-9 reduces in vitro invasion and angiogenesis in human microvascular endothelial cells. Int. J. Oncol. 25, 1407–1414 (2004)PubMed

44.

Y. Jiao, X. Feng, Y. Zhan, R. Wang, S. Zheng, W. Liu, X. Zeng, Matrix metalloproteinase-2 promotes αvβ3 integrin-mediated adhesion and migration of human melanoma cells by cleaving fibronectin. PLoS One 7, e41591 (2012). doi:10.1371/journal.pone.0041591

45.

J.M. Yang, Z. Xu, H. Wu, H. Zhu, X. Wu, W.N. Hait, Overexpression of extracellular matrix metalloproteinase inducer in multidrug resistant cancer cells. Mol. Cancer Res. 1, 420–427 (2003)

46.

F.Y. Zhang, Y. Hu, Z.Y. Que, P. Wang, Y.H. Liu, Z.H. Wang, Y.X. Xue, Shikonin inhibits the migration and invasion of human glioblastoma cells by targeting phosphorylated β-catenin and phosphorylated PI3K/Akt: A potential mechanism for the anti-glioma efficacy of a traditional Chinese herbal medicine. Int. J. Mol. Sci. 16, 23823–23848 (2015). doi:10.3390/ijms161023823 CrossRefPubMedPubMedCentral

47.

P.L. Wei, C.C. Tu, C.H. Chen, Y.S. Ho, C.T. Wu, H.Y. Su, W.Y. Chen, J.J. Liu, Y.J. Chang, Shikonin suppresses the migratory ability of hepatocellular carcinoma cells. J. Agric. Food Chem. 61, 8191–8197 (2013). doi:10.1021/jf4009586

Titel: Dual treatment with shikonin and temozolomide reduces glioblastoma tumor growth, migration and glial-to-mesenchymal transition
verfasst von: Diana Matias
Joana Balça-Silva
Luiz Gustavo Dubois
Bruno Pontes
Valéria Pereira Ferrer
Luciane Rosário
Anália do Carmo
Juliana Echevarria-Lima
Ana Bela Sarmento-Ribeiro
Maria Celeste Lopes
Vivaldo Moura-Neto
Publikationsdatum: 11.04.2017
Verlag: Springer Netherlands
Erschienen in: Cellular Oncology / Ausgabe 3/2017
Print ISSN: 2211-3428
Elektronische ISSN: 2211-3436
DOI: https://doi.org/10.1007/s13402-017-0320-1

Springer Medizin

Dual treatment with shikonin and temozolomide reduces glioblastoma tumor growth, migration and glial-to-mesenchymal transition

Abstract

Purpose

Methods

Results

Conclusion

Neu im Fachgebiet Pathologie

Molekularpathologische Untersuchungen im Wandel der Zeit

Vergleichende Pathologie in der onkologischen Forschung

Gastrointestinale Stromatumoren

Personalisierte Medizin in der Onkologie

Springer Medizin

Abstract

Purpose

Methods

Results

Conclusion

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