Increased invasive capacity of connexin43-overexpressing malignant glioma cells

Wei Zhang M.D., Ph.D.1, Chiedozie Nwagwu M.D.1, Duc Minh Le Ph.D.1, V. Wee Yong Ph.D.1, Hua Song M.D.1, and William T. Couldwell M.D., Ph.D.1
View More View Less
  • 1 Departments of Neurosurgery and Cell Biology & Anatomy, New York Medical College, Valhalla and New York, New York; Department of Neurosurgery, University of Utah, Salt Lake City, Utah; Neuro-Oncology Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland; and Departments of Oncology and Clinical Neurosciences, University of Calgary, Alberta, Canada
Restricted access

Purchase Now

USD  $45.00

JNS + Pediatrics - 1 year subscription bundle (Individuals Only)

USD  $515.00

JNS + Pediatrics + Spine - 1 year subscription bundle (Individuals Only)

USD  $612.00
Print or Print + Online

Object. Malignant glioma cells, similar to astrocytes, express connexin43 (Cx43) universally but at widely varied levels. Data from previous studies have demonstrated that malignant glioma cells form functional gap junction channels among themselves as well as with astrocytes and that such a communication has the potential to modulate the phenotypic characteristics of astrocytes. Recently, gap junctions have been demonstrated to play a role in the invasive phenotype of malignant gliomas. In this study, the authors have further investigated the motility and invasion ability of Cx43-overexpressing and Cx43-deficient malignant glioma cells.

Methods. Using a standard invasion system of a Matrigel transwell invasion chamber, the authors found that the number of Cx43-transfected C6 glioma cells (C6-Cx43 cells) migrating through the Matrigel-coated membrane was similar to that of mock-transfected control cells (C6-mock cells) during the first 24 hours, but increased significantly thereafter. When these cells were cocultured with astrocytes, the number of invading C6-Cx43 cells was more than threefold greater than the number of invading C6-mock cells. Results of an in vitro cell motility assay also demonstrated that C6-Cx43 cells were more motile and scatter-active than C6-mock cells. Furthermore, zymographic analysis of MMPs, an important determinant in glioma invasion, demonstrated that the amounts of MMP-2 and MMP-9 in culture medium collected from C6-Cx43 cells were orders of magnitude higher than those from C6-mock cells. In addition, BB-94, a synthetic MMP inhibitor, significantly inhibited C6-Cx43 cell invasion.

Conclusions. The overexpression of gap junction proteins in glioma cells and the intercellular communication between tumor and nontumor glia cells may play important roles in the facilitation of glioma cell invasion.

JNS + Pediatrics - 1 year subscription bundle (Individuals Only)

USD  $515.00

JNS + Pediatrics + Spine - 1 year subscription bundle (Individuals Only)

USD  $612.00
  • 1.

    Amberger VR, , Hensel T, & Ogata N, et al: Spreading and migration of human glioma and rat C6 cells on central nervous system myelin in vitro is correlated with tumor malignancy and involves a metalloproteolytic activity. Cancer Res 58:149158, 1998 Amberger VR, Hensel T, Ogata N, et al: Spreading and migration of human glioma and rat C6 cells on central nervous system myelin in vitro is correlated with tumor malignancy and involves a metalloproteolytic activity. Cancer Res 58:149–158, 1998

    • Search Google Scholar
    • Export Citation
  • 2.

    Bernstein JJ, , Goldberg WJ, & Laws ER Jr, et al: C6 glioma cell invasion and migration of rat brain after neural homografting: ultrastructure. Neurosurgery 26:622628, 1990 Bernstein JJ, Goldberg WJ, Laws ER Jr, et al: C6 glioma cell invasion and migration of rat brain after neural homografting: ultrastructure. Neurosurgery 26:622–628, 1990

    • Search Google Scholar
    • Export Citation
  • 3.

    Bernstein LR, & Liotta LA: Molecular mediators of interactions with extracellular matrix components in metastasis and angiogenesis. Curr Opin Oncol 6:106113, 1994 Bernstein LR, Liotta LA: Molecular mediators of interactions with extracellular matrix components in metastasis and angiogenesis. Curr Opin Oncol 6:106–113, 1994

    • Search Google Scholar
    • Export Citation
  • 4.

    Burger PC, & Vollmer RT: Histologic factors of prognostic significance in the glioblastoma multiforme. Cancer 46:11791186, 1980 Burger PC, Vollmer RT: Histologic factors of prognostic significance in the glioblastoma multiforme. Cancer 46:1179–1186, 1980

    • Search Google Scholar
    • Export Citation
  • 5.

    Chicoine MR, & Silbergeld DL: Assessment of brain tumor cell motility in vivo and in vitro. J Neurosurg 82:615622, 1995 Chicoine MR, Silbergeld DL: Assessment of brain tumor cell motility in vivo and in vitro. J Neurosurg 82:615–622, 1995

    • Search Google Scholar
    • Export Citation
  • 6.

    Chintala SK, , Fueyo J, & Gomez-Manzano C, et al: Adenovirus-mediated p16/CDKN2 gene transfer suppresses glioma invasion in vitro. Oncogene 15:20492057, 1997 Chintala SK, Fueyo J, Gomez-Manzano C, et al: Adenovirus-mediated p16/CDKN2 gene transfer suppresses glioma invasion in vitro. Oncogene 15:2049–2057, 1997

    • Search Google Scholar
    • Export Citation
  • 7.

    Chintala SK, , Sawaya R, & Aggarwal BB, et al: Induction of matrix metalloproteinase-9 requires a polymerized actin cytoskeleton in human malignant glioma cells. J Biol Chem 273:1354513551, 1998 Chintala SK, Sawaya R, Aggarwal BB, et al: Induction of matrix metalloproteinase-9 requires a polymerized actin cytoskeleton in human malignant glioma cells. J Biol Chem 273:13545–13551, 1998

    • Search Google Scholar
    • Export Citation
  • 8.

    Cotrina ML, , Lin JH, & Nedergaard M: Cytoskeletal assembly and ATP release regulate astrocytic calcium signaling. J Neurosci 18:87948804, 1998 Cotrina ML, Lin JH, Nedergaard M: Cytoskeletal assembly and ATP release regulate astrocytic calcium signaling. J Neurosci 18:8794–8804, 1998

    • Search Google Scholar
    • Export Citation
  • 9.

    Dermietzel R, & Spray DC: Gap junctions in the brain: where, what type, how many and why? Trends Neurosci 16:186192, 1993 Dermietzel R, Spray DC: Gap junctions in the brain: where, what type, how many and why? Trends Neurosci 16:186–192, 1993

    • Search Google Scholar
    • Export Citation
  • 10.

    Frosch BA, & Sloane BF: The role of proteolytic enzymes in brain tumor infiltration, in Mikkelsen T, , Bjerkvig R, & Laerum OD, et al (eds): Brain Tumor Invasion: Biological, Clinical and Therapeutic Considerations. New York: Wiley-Liss, Inc: 1998, pp 275300 Frosch BA, Sloane BF: The role of proteolytic enzymes in brain tumor infiltration, in Mikkelsen T, Bjerkvig R, Laerum OD, et al (eds): Brain Tumor Invasion: Biological, Clinical and Therapeutic Considerations. New York: Wiley-Liss, Inc: 1998, pp 275–300

    • Search Google Scholar
    • Export Citation
  • 11.

    Giese A, & Westphal M: Glioma invasion in the central nervous system. Neurosurgery 39:235252, 1996 Giese A, Westphal M: Glioma invasion in the central nervous system. Neurosurgery 39:235–252, 1996

    • Search Google Scholar
    • Export Citation
  • 12.

    Graeber SH, & Hulser DF: Connexin transfection induces invasive properties in HeLa cells. Exp Cell Res 243:142149, 1998 Graeber SH, Hulser DF: Connexin transfection induces invasive properties in HeLa cells. Exp Cell Res 243:142–149, 1998

    • Search Google Scholar
    • Export Citation
  • 13.

    Grobben B, , De Deyn PP, & Slegers H: Rat C6 glioma as experimental model system for the study of glioblastoma growth and invasion. Cell Tissue Res 310:257270, 2002 Grobben B, De Deyn PP, Slegers H: Rat C6 glioma as experimental model system for the study of glioblastoma growth and invasion. Cell Tissue Res 310:257–270, 2002

    • Search Google Scholar
    • Export Citation
  • 14.

    Horwitz AR, & Parsons JT: Cell migration—movin' on. Science 286:11021103, 1999 Horwitz AR, Parsons JT: Cell migration—movin' on. Science 286:1102–1103, 1999

    • Search Google Scholar
    • Export Citation
  • 15.

    Kaya M, , Yoshida K, & Higashino F, et al: A single ets-related transcription factor, E1AF, confers invasive phenotype on human cancer cells. Oncogene 12:221227, 1996 Kaya M, Yoshida K, Higashino F, et al: A single ets-related transcription factor, E1AF, confers invasive phenotype on human cancer cells. Oncogene 12:221–227, 1996

    • Search Google Scholar
    • Export Citation
  • 16.

    Knott JC, , Mahesparan R, & Garcia-Cabrera I, et al: Stimulation of extracellular matrix components in the normal brain by invading glioma cells. Int J Cancer 75:864872, 1998 Knott JC, Mahesparan R, Garcia-Cabrera I, et al: Stimulation of extracellular matrix components in the normal brain by invading glioma cells. Int J Cancer 75:864–872, 1998

    • Search Google Scholar
    • Export Citation
  • 17.

    Lauffenburger DA, & Horwitz AF: Cell migration: a physically integrated molecular process. Cell 84:359369, 1996 Lauffenburger DA, Horwitz AF: Cell migration: a physically integrated molecular process. Cell 84:359–369, 1996

    • Search Google Scholar
    • Export Citation
  • 18.

    Lin JHC, , Takano T, & Cotrina ML, et al: Connexin 43 enhances the adhesivity and mediates the invasion of malignant glioma cells. J Neurosci 22:43024311, 2002 Lin JHC, Takano T, Cotrina ML, et al: Connexin 43 enhances the adhesivity and mediates the invasion of malignant glioma cells. J Neurosci 22:4302–4311, 2002

    • Search Google Scholar
    • Export Citation
  • 19.

    Liotta LA, & Stetler-Stevenson WG: Metalloproteinases and cancer invasion. Semin Cancer Biol 1:99106, 1990 Liotta LA, Stetler-Stevenson WG: Metalloproteinases and cancer invasion. Semin Cancer Biol 1:99–106, 1990

    • Search Google Scholar
    • Export Citation
  • 20.

    Nagano N, , Sasaki H, & Aoyagi M, et al: Invasion of experimental rat brain tumor: early morphological changes following microinjection of C6 glioma cells. Acta Neuropathol 86:117125, 1993 Nagano N, Sasaki H, Aoyagi M, et al: Invasion of experimental rat brain tumor: early morphological changes following microinjection of C6 glioma cells. Acta Neuropathol 86:117–125, 1993

    • Search Google Scholar
    • Export Citation
  • 21.

    Naus CC, , Zhu D, & Todd SD, et al: Characteristics of C6 glioma cells overexpressing a gap junction protein. Cell Mol Neurobiol 12:163175, 1992 Naus CC, Zhu D, Todd SD, et al: Characteristics of C6 glioma cells overexpressing a gap junction protein. Cell Mol Neurobiol 12:163–175, 1992

    • Search Google Scholar
    • Export Citation
  • 22.

    Naus CCG, , Bechberger JF, & Bond SL: Effect of gap junctional communication on glioma cell function, in Spray DC, & Dermietzel R (eds): Neuroscience Intelligence Unit: Gap Junctions in the Nervous System. Austin, TX: RG Landes, 1996, pp 193202 Naus CCG, Bechberger JF, Bond SL: Effect of gap junctional communication on glioma cell function, in Spray DC, Dermietzel R (eds): Neuroscience Intelligence Unit: Gap Junctions in the Nervous System. Austin, TX: RG Landes, 1996, pp 193–202

    • Search Google Scholar
    • Export Citation
  • 23.

    Paulus W, , Baur I, & Schuppan D, et al: Characterization of integrin receptors in normal and neoplastic human brain. Am J Pathol 143:154163, 1993 Paulus W, Baur I, Schuppan D, et al: Characterization of integrin receptors in normal and neoplastic human brain. Am J Pathol 143:154–163, 1993

    • Search Google Scholar
    • Export Citation
  • 24.

    Pedersen PH, , Marienhagen K, & Mork S, et al: Migratory pattern of fetal rat brain cells and human glioma cells in the adult rat brain. Cancer Res 53:51585165, 1993 Pedersen PH, Marienhagen K, Mork S, et al: Migratory pattern of fetal rat brain cells and human glioma cells in the adult rat brain. Cancer Res 53:5158–5165, 1993

    • Search Google Scholar
    • Export Citation
  • 25.

    Rao JS, , Steck PA, & Mohanam S, et al: Elevated levels of M(r) 92,000 type IV collagenase in human brain tumors. Cancer Res 53 (Suppl 10):22082211, 1993 Rao JS, Steck PA, Mohanam S, et al: Elevated levels of M(r) 92,000 type IV collagenase in human brain tumors. Cancer Res 53 (Suppl 10):2208–2211, 1993

    • Search Google Scholar
    • Export Citation
  • 26.

    Salazar OM, & Rubin P: The spread of glioblastoma multiforme as a determining factor in the radiation treated volume. Int J Radiat Oncol Biol Phys 1:627637, 1976 Salazar OM, Rubin P: The spread of glioblastoma multiforme as a determining factor in the radiation treated volume. Int J Radiat Oncol Biol Phys 1:627–637, 1976

    • Search Google Scholar
    • Export Citation
  • 27.

    Sawaya RE, , Yamamoto M, & Gokaslan ZL, et al: Expression and localization of 72 kDa type IV collagenase (MMP-2) in human malignant gliomas in vivo. Clin Exp Metastasis 14:3542, 1996 Sawaya RE, Yamamoto M, Gokaslan ZL, et al: Expression and localization of 72 kDa type IV collagenase (MMP-2) in human malignant gliomas in vivo. Clin Exp Metastasis 14:35–42, 1996

    • Search Google Scholar
    • Export Citation
  • 28.

    Shinoura N, , Chen L, & Wani MA, et al: Protein and messenger RNA expression of connexin43 in astrocytomas: implications in brain tumor gene therapy. J Neurosurg 84:839846, 1996 Shinoura N, Chen L, Wani MA, et al: Protein and messenger RNA expression of connexin43 in astrocytomas: implications in brain tumor gene therapy. J Neurosurg 84:839–846, 1996

    • Search Google Scholar
    • Export Citation
  • 29.

    Uhm JH, , Dooley NP, & Villemure JG, et al: Glioma invasion in vitro: regulation by matrix metalloprotease-2 and protein kinase C. Clin Exp Metastasis 14:421433, 1996 Uhm JH, Dooley NP, Villemure JG, et al: Glioma invasion in vitro: regulation by matrix metalloprotease-2 and protein kinase C. Clin Exp Metastasis 14:421–433, 1996

    • Search Google Scholar
    • Export Citation
  • 30.

    Yamane Y, , Shiga H, & Asou H, et al: Dynamics of astrocyte adhesion as analyzed by a combination of atomic force microscopy and immuno-cytochemistry: the involvement of actin filaments and connexin 43 in the early stage of adhesion. Arch Histol Cytol 62:355361, 1999 Yamane Y, Shiga H, Asou H, et al: Dynamics of astrocyte adhesion as analyzed by a combination of atomic force microscopy and immuno-cytochemistry: the involvement of actin filaments and connexin 43 in the early stage of adhesion. Arch Histol Cytol 62:355–361, 1999

    • Search Google Scholar
    • Export Citation
  • 31.

    Yong VW, , Krekoski CA, & Forsyth PA, et al: Matrix metalloproteinases and diseases of the CNS. Trends Neurosci 21:7580, 1998 Yong VW, Krekoski CA, Forsyth PA, et al: Matrix metalloproteinases and diseases of the CNS. Trends Neurosci 21:75–80, 1998

    • Search Google Scholar
    • Export Citation
  • 32.

    Zhang W, , Couldwell WT, & Simard MF, et al: Direct gap junction communication between malignant glioma cells and astrocytes. Cancer Res 59:19942003, 1999 Zhang W, Couldwell WT, Simard MF, et al: Direct gap junction communication between malignant glioma cells and astrocytes. Cancer Res 59:1994–2003, 1999

    • Search Google Scholar
    • Export Citation
  • 33.

    Zhang W, , Law RE, & Hinton DR, et al: Inhibition of human malignant glioma cell motility and invasion in vitro by hypericin, a potent protein kinase C inhibitor. Cancer Lett 120:3138, 1997 Zhang W, Law RE, Hinton DR, et al: Inhibition of human malignant glioma cell motility and invasion in vitro by hypericin, a potent protein kinase C inhibitor. Cancer Lett 120:31–38, 1997

    • Search Google Scholar
    • Export Citation
  • 34.

    Zhu D, , Kidder GM, & Caveney S, et al: Growth retardation in glioma cells cocultured with cells overexpressing a gap junction protein. Proc Natl Acad Sci USA 89:1021810221, 1992 Zhu D, Kidder GM, Caveney S, et al: Growth retardation in glioma cells cocultured with cells overexpressing a gap junction protein. Proc Natl Acad Sci USA 89:10218–10221, 1992

    • Search Google Scholar
    • Export Citation

Metrics

All Time Past Year Past 30 Days
Abstract Views 606 158 7
Full Text Views 246 5 0
PDF Downloads 115 4 0
EPUB Downloads 0 0 0