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Proc Natl Acad Sci U S A. Mar 1, 1991; 88(5): 1883–1887.
PMCID: PMC51130

Transfection of C6 glioma cells with connexin 43 cDNA: analysis of expression, intercellular coupling, and cell proliferation.

Abstract

C6 glioma cells express low levels of the gap junction protein connexin 43 and its mRNA and display very weak dye coupling. When implanted into the rat cerebrum, these cells quickly give rise to a large glioma. To investigate the role of gap junctions in the tumor characteristics of these cells, we have used Lipofectin-mediated transfection to introduce a full-length cDNA encoding connexin 43. Several transfected clones were obtained that exhibited various amounts of connexin 43 mRNA transcribed from the inserted cDNA. Immunocytochemical analysis revealed an increase in the amount of connexin 43 immunoreactivity in the transfected cells, being localized at areas of intercellular contact as well as in the cytoplasm. The level of dye coupling was also assessed and found to correlate with the amount of connexin 43 mRNA. When cell proliferation was followed over several days, cells expressing the transfected cDNA grew more slowly than non-transfected cells. These transfected cells will be useful in examining the role of gap junctions in tumorigenesis.

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  • Klaunig JE, Ruch RJ. Role of inhibition of intercellular communication in carcinogenesis. Lab Invest. 1990 Feb;62(2):135–146. [PubMed]
  • Guthrie SC, Gilula NB. Gap junctional communication and development. Trends Neurosci. 1989 Jan;12(1):12–16. [PubMed]
  • Stevenson BR, Paul DL. The molecular constituents of intercellular junctions. Curr Opin Cell Biol. 1989 Oct;1(5):884–891. [PubMed]
  • Beyer EC, Paul DL, Goodenough DA. Connexin family of gap junction proteins. J Membr Biol. 1990 Jul;116(3):187–194. [PubMed]
  • Bräuner T, Schmid A, Hülser DF. Tumor cell invasion and gap junctional communication. I. Normal and malignant cells confronted in monolayer cultures. Invasion Metastasis. 1990;10(1):18–30. [PubMed]
  • Tiffany-Castiglioni E, Neck KF, Caceci T. Glial culture on artificial capillaries: electron microscopic comparisons of C6 rat glioma cells and rat astroglia. J Neurosci Res. 1986;16(2):387–396. [PubMed]
  • Benda P, Lightbody J, Sato G, Levine L, Sweet W. Differentiated rat glial cell strain in tissue culture. Science. 1968 Jul 26;161(3839):370–371. [PubMed]
  • Beyer EC, Paul DL, Goodenough DA. Connexin43: a protein from rat heart homologous to a gap junction protein from liver. J Cell Biol. 1987 Dec;105(6 Pt 1):2621–2629. [PMC free article] [PubMed]
  • Eliyahu D, Michalovitz D, Oren M. Overproduction of p53 antigen makes established cells highly tumorigenic. Nature. 1985 Jul 11;316(6024):158–160. [PubMed]
  • Felgner PL, Gadek TR, Holm M, Roman R, Chan HW, Wenz M, Northrop JP, Ringold GM, Danielsen M. Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure. Proc Natl Acad Sci U S A. 1987 Nov;84(21):7413–7417. [PMC free article] [PubMed]
  • Chirgwin JM, Przybyla AE, MacDonald RJ, Rutter WJ. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. [PubMed]
  • Feinberg AP, Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. [PubMed]
  • Southern EM. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. [PubMed]
  • Safranyos RG, Caveney S. Rates of diffusion of fluorescent molecules via cell-to-cell membrane channels in a developing tissue. J Cell Biol. 1985 Mar;100(3):736–747. [PMC free article] [PubMed]
  • Beyer EC, Kistler J, Paul DL, Goodenough DA. Antisera directed against connexin43 peptides react with a 43-kD protein localized to gap junctions in myocardium and other tissues. J Cell Biol. 1989 Feb;108(2):595–605. [PMC free article] [PubMed]
  • Loewenstein WR. Junctional intercellular communication and the control of growth. Biochim Biophys Acta. 1979 Feb 4;560(1):1–65. [PubMed]
  • Steinsvåg SK, Laerum OD, Bjerkvig R. Interaction between rat glioma cells and normal rat brain tissue in organ culture. J Natl Cancer Inst. 1985 May;74(5):1095–1104. [PubMed]
  • Dermietzel R, Traub O, Hwang TK, Beyer E, Bennett MV, Spray DC, Willecke K. Differential expression of three gap junction proteins in developing and mature brain tissues. Proc Natl Acad Sci U S A. 1989 Dec;86(24):10148–10152. [PMC free article] [PubMed]
  • Dupont E, el Aoumari A, Roustiau-Sévère S, Briand JP, Gros D. Immunological characterization of rat cardiac gap junctions: presence of common antigenic determinants in heart of other vertebrate species and in various organs. J Membr Biol. 1988 Sep;104(2):119–128. [PubMed]
  • Paul DL. Molecular cloning of cDNA for rat liver gap junction protein. J Cell Biol. 1986 Jul;103(1):123–134. [PMC free article] [PubMed]
  • el Aoumari A, Fromaget C, Dupont E, Reggio H, Durbec P, Briand JP, Böller K, Kreitman B, Gros D. Conservation of a cytoplasmic carboxy-terminal domain of connexin 43, a gap junctional protein, in mammal heart and brain. J Membr Biol. 1990 May;115(3):229–240. [PubMed]
  • Naus CC, Belliveau DJ, Bechberger JF. Regional differences in connexin32 and connexin43 messenger RNAs in rat brain. Neurosci Lett. 1990 Apr 6;111(3):297–302. [PubMed]
  • Yamamoto T, Shiosaka S, Whittaker ME, Hertzberg EL, Nagy JI. Gap junction protein in rat hippocampus: light microscope immunohistochemical localization. J Comp Neurol. 1989 Mar 8;281(2):269–281. [PubMed]
  • Yamamoto T, Ochalski A, Hertzberg EL, Nagy JI. LM and EM immunolocalization of the gap junctional protein connexin 43 in rat brain. Brain Res. 1990 Feb 5;508(2):313–319. [PubMed]
  • Nicolson GL, Gallick GE, Dulski KM, Spohn WH, Lembo TM, Tainsky MA. Lack of correlation between intercellular junctional communication, p21rasEJ expression, and spontaneous metastatic properties of rat mammary cells after transfection with c-H-rasEJ or neo genes. Oncogene. 1990 May;5(5):747–753. [PubMed]
  • Eghbali B, Kessler JA, Spray DC. Expression of gap junction channels in communication-incompetent cells after stable transfection with cDNA encoding connexin 32. Proc Natl Acad Sci U S A. 1990 Feb;87(4):1328–1331. [PMC free article] [PubMed]
  • Fishman GI, Spray DC, Leinwand LA. Molecular characterization and functional expression of the human cardiac gap junction channel. J Cell Biol. 1990 Aug;111(2):589–598. [PMC free article] [PubMed]
  • Laerum OD, Hülser DF, Rajewsky MF. Electrophysiological properties of ethylnitrosourea-induced, neoplastic neurogenic rat cell lines cultured in vitro and in vivo. Cancer Res. 1976 Jul;36(7 Pt 1):2153–2161. [PubMed]

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