Biochemistry (Moscow)

, Volume 81, Issue 6, pp 628–635 | Cite as

Connection between proliferation rate and temozolomide sensitivity of primary glioblastoma cell culture and expression of YB-1 and LRP/MVP

  • N. I. MoiseevaEmail author
  • O. Yu. Susova
  • A. A. Mitrofanov
  • D. Yu. Panteleev
  • G. V. Pavlova
  • N. A. Pustogarov
  • A. A. Stavrovskaya
  • E. Yu. RybalkinaEmail author


Glioblastomas (GBL) are the most common and aggressive brain tumors. They are distinguished by high resistance to radiation and chemotherapy. To find novel approaches for GBL classification, we obtained 16 primary GBL cell cultures and tested them with real-time PCR for mRNA expression of several genes (YB-1, MGMT, MELK, MVP, MDR1, BCRP) involved in controlling cell proliferation and drug resistance. The primary GBL cultures differed in terms of proliferation rate, wherein a group of GBL cell cultures with low proliferation rate demonstrated higher resistance to temozolomide. We found that GBL primary cell cultures characterized by high proliferation rate and lower resistance to temozolomide expressed higher mRNA level of the YB-1 and MDR1 genes, whereas upregulated expression of MVP/LRP mRNA was a marker in the group of GBL with low proliferation rate and high resistance. A moderate correlation between expression of YB-1 and MELK as well as YB-1 and MDR1 was found. In the case of YB-1 and MGMT expression, no correlation was found. A significant negative correlation was revealed between mRNA expression of MVP/LRP and MELK, MDR1, and BCRP. No correlation in expression of YB-1 and MVP/LRP genes was observed. It seems that mRNA expression of YB-1 and MVP/LRP may serve as a marker for GBL cell cultures belonging to distinct groups, each of which is characterized by a unique pattern of gene activity.

Key words

glioblastoma primary cell culture temozolomide drug resistance proliferation YB-1 MVP 


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  1. 1.
    Schwartzbaum, J. A., Fisher, J. L., Aldape, K. D., and Wrensch, M. (2006) Epidemiology and molecular pathology of glioma, Nat. Clin. Pract. Neurol., 2, 494–503.CrossRefPubMedGoogle Scholar
  2. 2.
    Stupp, R., Hegi, M. E., Mason, W. P., Van den Bent, M. J., Taphoornm, M. J., Janzer, R. C., Ludwin, S. K., Allgeierm, A., Fisher, B., Belanger, K., Hau, P., Brandes, A. A., Gijtenbeekm, J., Marosi, C., Vecht, C. J., Mokhtari, K., Wesseling, P., Villa, S., Eisenhauer, E., Gorlia, T., Weller, M., Lacombe, D., Cairncross, J. G., Mirimanoff, R. O., Stupp, R., Hegi, M. E., and Mason, W. P. (2009) Effects of radiotherapy with concomitant and adjuvant temozolomide versus radiotherapy alone on survival in glioblastoma in a randomized phase III study: 5-year analysis of the EORTC-NCIC trial, Lancet Oncol., 10, 459–466.CrossRefPubMedGoogle Scholar
  3. 3.
    Villalva, C., Cortes, U., Wager, M., Tourani, J. M., Rivet, P., Marquant, C., Martin, S., Turhan, A. G., and KarayanTapon, L. (2012) O6-Methylguanine-methyltransferase (MGMT) promoter methylation status in glioma stem-like cells is correlated to temozolomide sensitivity under differentiation-promoting conditions, Int. J. Mol. Sci., 13, 69836994.CrossRefGoogle Scholar
  4. 4.
    Parker, N. R., Khong, P., Parkinson, J. F., Howell, V. M., and Wheeler, H. R. (2015) Molecular heterogeneity in glioblastoma: potential clinical implications, Front. Oncol., 5, 1–9.Google Scholar
  5. 5.
    Stavrovskaya, A. A., and Gens, G. P. (2014) Some new aspects of investigating multi-drug resistance in tumor cells, Uspekhi Mol. Onkol., 1, 5–11.Google Scholar
  6. 6.
    Spiegl-Kreinecker, S., Buchroithner, J., Elbling, L., Steiner, E., Wurm, G., Bodenteich, A., Fischer, J., Micksche, M., and Berger, W. (2002) Expression and functional activity of the ABC-transporter proteins P-glycoprotein and multidrug-resistance protein 1 in human brain tumor cells and astrocytes, J. Neurooncol., 57, 27–36.CrossRefPubMedGoogle Scholar
  7. 7.
    Abe, T., Mori, T., Wakabayashi, Y., Nakagawa, M., Cole, S. P., Koike, K., Kuwano, M., and Hori, S. (1998) Expression of multidrug resistance protein gene in patients with glioma after chemotherapy, J. Neurooncol., 40, 11–18.CrossRefPubMedGoogle Scholar
  8. 8.
    Calatozzolo, C., Gelati, M., Ciusani, E., Sciacca, F. L., Pollo, B., Cajola, L., Marras, C., Silvani, A., VitellaroZuccarello, L., Croci, D., Boiardi, A., and Salmaggi, A. (2005) Expression of drug resistance proteins Pgp, MRP1, MRP3, MRP5 and GST-pi in human glioma, J. Neurooncol., 74, 113–121.Google Scholar
  9. 9.
    Scheffer, G. L., Wijngaard, P. L., Flens, M. J., Izquierdo, M. A., Slovak, M. L., Pinedo, H. M., Meijer, C. J., Clevers, H. C., and Scheper, R. J. (1995) The drug resistance-related protein LRP is the human major vault protein, Nat. Med., 1, 578–582.CrossRefPubMedGoogle Scholar
  10. 10.
    Lotsch, D., Steiner, E., Holzmann, K., Spiegl-Kreinecker, S., Pirker, C., Hlavaty, J., Petznek, H., Hegedus, B., Garay, T., Mohr, T., Sommergruber, W., Grusch, M., and Berger, W. (2013) Major vault protein supports glioblastoma survival and migration by upregulating the EGFR/PI3K signaling axis, Oncotarget, 4, 1904–1918.CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Lu, C., and Shervington, A. (2008) Chemoresistance in gliomas, Mol. Cell. Biochem., 312, 71–80.CrossRefPubMedGoogle Scholar
  12. 12.
    Navarro, L., Gil-Benso, R., Megias, J., Munoz-Hidalgo, L., San-Miguel, T., Callaghan, R. C., Gonzalez-Darder, J. M., Lopez-Gines, C., and Cerda-Nicolas, M. J. (2015) Alteration of major vault protein in human glioblastoma and its relation with EGFR and PTEN status, Neuroscience, 25, 243–251.CrossRefGoogle Scholar
  13. 13.
    Eliseeva, I. A., Kim, E. R., Gur’yanov, S. G., Ovchinnikov, L. P., and Lyabin, D. N. (2011) Y-box-binding protein 1 (YB-1) and its functions, Biochemistry (Moscow), 76, 14021433.CrossRefGoogle Scholar
  14. 14.
    Faury, D., Nantel, A., Dunn, S. E., Guiot, M. C., Haque, T., Hauser, P., Garami, M., Bognar, L., Hanzely, Z., Liberski, P. P., Lopez-Aguilar, E., Valera, E. T., Tone, L. G., Carret, A. S., Del Maestro, R. F., Gleave, M., Montes, J. L., Pietsch, T., Albrecht, S., and Jabado, N. (2007) Molecular profiling identifies prognostic subgroups of pediatric glioblastoma and shows increased YB-1 expression in tumors, J. Clin. Oncol., 25, 1196–1208.CrossRefPubMedGoogle Scholar
  15. 15.
    Gao, Y., Fotovati, A., Lee, C., Wang, M., Cote, G., Guns, E., Toyota, B., Faury, D., Jabado, N., and Dunn, S. E. (2009) Inhibition of Y-box binding protein-1 slows the growth of glioblastoma multiforme and sensitizes to temozolomide methyltransferase, Mol. Cancer Ther., 8, 32763284.Google Scholar
  16. 16.
    Nakano, I., Masterman-Smith, M., Saigusa, K., Paucar, A. A., Horvath, S., Shoemaker, L., Watanabe, M., Negro, A., Bajpai, R., Howes, A., Lelievre, V., Waschek, J. A., Lazareff, J. A., Freije, W. A., Liau, L. M., Gilbertson, R. J., Cloughesy, T. F., Geschwind, D. H., Nelson, S. F., Mischel, P. S., Terskikh, A. V., and Kornblum, H. I. (2008) Maternal embryonic leucine zipper kinase is a key regulator of the proliferation of malignant brain tumors, including brain tumor stem cells, J. Neurosci. Res., 86, 48–60.PubMedGoogle Scholar
  17. 17.
    Joshi, K., Banasavadi-Siddegowda, Y., Mo, X., Kim, S. H., Mao, P., Kig, C., Nardini, D., Sobol, R. W., Chow, L. M., Kornblum, H. I., Waclaw, R., Beullens, M., and Nakano, I. (2013) MELK-dependent FOXM1 phosphorylation is essential for proliferation of glioma stem cells, Stem Cells, 31, 1051–1063.CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Ganguly, R., Hong, C. S., Smith, L. G., Kornblum, H. I., and Nakano, I. (2014) Maternal embryonic leucine zipper kinase: key kinase for stem cell phenotype in glioma and other cancers, Mol. Cancer Ther., 13, 1393–1398.CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Stein, U., Bergmann, S., Scheffer, G. L., Scheper, R. J., Royer, H. D., Schlag, P. M., and Walther, W. (2005) YB-1 facilitates basal and 5-fluorouracil-inducible expression of the human major vault protein (MVP) gene, Oncogene, 24, 3606–3618.CrossRefPubMedGoogle Scholar
  20. 20.
    Oda, Y., Ohishi, Y., Basaki, Y., Kobayashi, H., Hirakawa, T., Wake, N., Ono, M., Nishio, K., Kuwano, M., and Tsuneyoshi, M. (2007) Prognostic implications of the nuclear localization of Y-box-binding protein-1 and CXCR4 expression in ovarian cancer: their correlation with activated Akt, LRP/MVP and P-glycoprotein expression, Cancer Sci., 98, 1020–1026.PubMedGoogle Scholar
  21. 21.
    Vayman, A. V., Stromskaya, T. P., Rybalkina, E. Yu., Sorokin, A. V., Gur’yanov, S. G., Zabotina, T. N., Mechetner, E. B., Ovchinnikov, L. P., and Stavrovskaya, A. A. (2006) Intracellular localization and content of YB-1 protein in multidrug resistant tumor cells, Biochemistry (Moscow), 71, 146–154.CrossRefGoogle Scholar
  22. 22.
    Kuwano, M., Oda, Y., Izumi, H., Yang, S. J., Uchiumi, T., Iwamoto, Y., Toi, M., Fujii, T., Yamana, H., Kinoshita, H., Kamura, T., Tsuneyoshi, M., Yasumoto, K., and Kohno, K. (2004) The role of nuclear Y-box binding protein 1 as a global marker in drug resistance, Mol. Cancer Ther., 3, 1485–1492.PubMedGoogle Scholar
  23. 23.
    Stavrovskaya, A. A., and Stromskaya, T. P. (2008) Transport proteins of the ABC family and multidrug resistance of tumor cells, Biochemistry (Moscow), 73, 735–750.CrossRefGoogle Scholar
  24. 24.
    Stavrovskaya, A., Stromskaya, T., Rybalkina, E., Moiseeva, N., Vaiman, A., Guryanov, S., Ovchinnikov, L., and Guens, G. (2012) YB-1 protein and multidrug resistance of tumor cells, Curr. Signal Trans. Ther., 7, 237–246.CrossRefGoogle Scholar
  25. 25.
    Fotovati, A., Abu-Ali, S., Wang, P. S., Deleyrolle, L. P., Lee, C., Triscott, J., Chen, J. Y., Franciosi, S., Nakamura, Y., Sugita, Y., Uchiumi, T., Kuwano, M., Leavitt, B. R., Singh, S. K., Jury, A., Jones, C., Wakimoto, H., Reynolds, B. A., Pallen, C. J., and Dunn, S. E. (2011) YB-1 bridges neural stem cells and brain tumor-initiating cells via its roles in differentiation and cell growth, Cancer Res., 71, 5569–5578.CrossRefPubMedGoogle Scholar
  26. 26.
    Steiner, E., Holzmann, K., Elbling, L., Micksche, M., and Berger, W. (2006) Cellular functions of vaults and their involvement in multidrug resistance, Curr. Drug Targets, 7, 923–934.CrossRefPubMedGoogle Scholar
  27. 27.
    Berger, W., Steiner, E., Grusch, M., Elbling, L., and Micksche, M. (2009) Vaults and the major vault protein: novel roles in signal pathway regulation and immunity, Cell. Mol. Life Sci., 66, 43–61.CrossRefPubMedGoogle Scholar
  28. 28.
    Lotsch, D., Steiner, E., Holzmann, K., Spiegl-Kreinecker, S., Pirker, C., Hlavaty, J., Petznek, H., Hegedus, B., Garay, T., Mohr, T., Sommergruber, W., Grusch, M., and Berger, W. (2013) Major vault protein supports glioblastoma survival and migration by upregulating the EGFR/PI3K signaling axis, Oncotarget, 11, 1904–1918.CrossRefGoogle Scholar
  29. 29.
    Henriquez-Hernandez, L. A., Moreno, M., Rey, A., Lloret, M., and Lara, P. C. (2012) MVP expression in the prediction of clinical outcome of locally advanced oral squamous cell carcinoma patients treated with radiotherapy, Radiat. Oncol., 7, doi: 10.1186/1748-717X-7-147.Google Scholar

Copyright information

© Pleiades Publishing, Ltd. 2016

Authors and Affiliations

  • N. I. Moiseeva
    • 1
    Email author
  • O. Yu. Susova
    • 1
  • A. A. Mitrofanov
    • 1
  • D. Yu. Panteleev
    • 2
  • G. V. Pavlova
    • 2
  • N. A. Pustogarov
    • 2
  • A. A. Stavrovskaya
    • 1
  • E. Yu. Rybalkina
    • 1
    Email author
  1. 1.Blokhin Russian Cancer Research CenterMinistry of Health of the Russian FederationMoscowRussia
  2. 2.Institute of Gene BiologyRussian Academy of SciencesMoscowRussia

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