Abstract
Two weeks after the inoculation of 1.5 × 105 9L glioma cells into the rat brain, the uptake of radiolabelled drugs into the brain and the experimental 9L glioma during the first cerebral circulation was measured with a liquid scintillation counter and analyzed by the method of Oldendorf (1970). The expression of P-glycoprotein, which is known to be associated with the efflux of drugs, was also studied, using anti-P-glycoprotein monoclonal antibody, C-219. Furthermore, the ultrastructure of brain capillaries, tumor vessels, and glioma cells was studied by conventional and immunoelectron microscopy. Sucrose (control), the transport of which through the blood-brain barrier is known to be negligible, accumulated to fivefold higher levels in the tumor than in normal brain. Ranimustine (MCNU), 5-fluorouracil (5-FU), and doxorubicin showed little accumulation in the normal brain, whereas nimustine (ACNU) showed an increased accumulation. MCNU and doxorubicin showed negligible accumulation in the glioma cells despite diffusion into the tumor interstitial space. In contrast, ACNU and 5-FU showed an increased accumulation in tumor cells. The accumulation of 5-FU in the cultured 9L glioma cells was decreased by ATP inhibitors or by low temperature. Although both brain capillary endothelial cells and glioma cell membrane were immunohisto-chemically positive for P-glycoprotein, the tumor vasculature showed low expression of P-glycoprotein. The endothelial cells of tumor vessels ultrastructurally showed increased fenestrations, swelling, and disrupted junctions. Accordingly, it is suggested that hydrophobic drugs such as doxorubicin, being pumped out by P-glycoprotein, do not accumulate in 9L glioma cells as do other lipophilic drugs such as ACNU, or drugs such as 5-FU, which accumulate by a carrier-mediated mechanism.
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References
Arena ED, Alessandro N, Dusonchet L, Gebbia N, Gerbasi F, Sanguedolce R, Rausa L (1972) Influence of pharmacokinetic variations on the pharmacological properties of Adriamycin. In: Carter SK, DiMarco A, Ghione M, Krakoff IH, Mathé G (eds) International symposium on Adriamycin. Springer, Berlin Heidelberg New York, pp 96–116
Benda P, Someda K, Messer J, Sweet WH (1971) Morphological and immunochemical studies of rat glial tumors and clonal strains propagated in culture. J Neurosurg 34:310–323
Bradbury MWB, Patlak CS, Oldendorf WH (1975) Analysis of brain uptake and loss of radiotracers after intracarotid injection. Am J Physiol 229:1110–1115
Brightman MW, Klatzo I, Olsson Y, Reese TS (1970) The blood-brain barrier to proteins under normal and pathological conditions. J Neurol Sci 10:215–239
Brightman MW, Reese TS, Vick NA, Bigner DD (1971) A mechanism underlying the lack of a blood-brain barrier to peroxidase in virally induced brain tumors (abstract). J Neuropathol Exp Neurol 30:139–140
Chan HS, Thorner PS, Haddad G, Ling V (1990) Immunohistochemical detection of P-glycoprotein: prognostic correlation in soft tissue sarcoma of childhood. J Clin Oncol 8:689–704
Chen YN, Mickley LA, Schwartz AM, Acton EM, Hwang J, Fojo AT (1990) Characterization of Adriamycin-resistant human breast cancer cells which display overexpression of a novel resistance-related membrane protein. J Biol Chem 265:10073–10080
Cordon-Cardo C, O'Brien JP, Casals D, Rittman-Grauer L, Biedler JL, Melamed MR, Bertino J (1989) Multidrug-resistance gene (P-glycoprotein) is expressed by endothelial cells at blood-brain barrier sites. Proc Natl Acad Sci USA 86:695–698
Fojo AT, Ueda K, Slamon DJ, Poplack DG, Gottesman MM, Pastan I (1987) Expression of a multidrug-resistance gene in human tumors and tissues. Proc Natl Acad Sci USA 84:265–269
Goldstein LJ, Fojo AT, Ueda K, Crist W, Green A, Brodeur G, Pastan I, Gottesman MM (1990) Expression of the multidrug resistance, MDR1, gene in neuroblastomas. J Clin Oncol 8:128–136
Gottesman MM, Pastan I (1988) The multidrug transporter, a double-edged sword. J Biol Chem 263:12163–12166
Harada K, Kiya K, Uozumi T (1981) Pharmacokinetics of a new water-soluble nitrosourea derivative (ACNU) in human gliomas. Surg Neurol 15:410–414
Hasegawa H, Ushio Y, Hayakawa T, Yamada K, Mogami H (1983) Changes of the blood-brain barrier in experimental metastatic brain tumors. J Neurosurg 59:304–310
Juliano RL, Ling VA (1976) A surface glycoprotein modulating drug permeability in Chinese hamster ovary cell mutants. Biochim Biophys Acta 455:152–162
Kartner N, Evernden-Porelle D, Bradley G, Ling V (1985) Detection of P-glycoprotein in multidrug-resistant cell lines by monoclonal antibodies. Nature 316:820–823
Kimura K, Fujita H, Sakai Y (1972) Blood levels, tissue distribution and clinical effects of Adriamycin. In: Carter SK, DiMarco A, Ghione M, Krakoff IH, Mathé G (eds) International symposium on Adriamycin. Springer. Berlin Heidelberg New York, pp 124–134
Levin VA, Landahl HD, Freeman-Dove MA (1976) The application of brain capillary permeability coefficient measurements to pathological conditions and the selection of agents which cross the blood-brain barrier. J Pharmacokinet Biopharm 4:499–519
Mori T (1988) The pharmacokinetics of ACNU in patients with malignant brain tumor. In: Suzuki J (ed) Treatment of glioma. Springer, Berlin Heidelberg New York, pp 109–116
Moscow JA, Fairchild CR, Madden MJ, Ransom DT, Wieand HS, O'Brien EE, Poplack DG, Cossman J, Myers CE, Cowan KH (1989) Expression of anionic glutathione-S-transferase and P-glycoprotein genes in human tissues and tumors. Cancer Res 49:1422–1428
Nabors MW, Griffin CA, Zehnbauer BA, Hruban RH, Phillips PC, Grossman SA, Brem H, Colvin OM (1991) Multidrug-resistance gene (MDR1) expression in human brain tumors. J Neurosurg 75:941–946
Oldendorf WH (1970) Measurement of brain uptake of radiolabeled substances using a tritiated water internal standard. Brain Res 24:372–376
Oldendorf WH (1974) Blood-brain barrier permeability to drugs. Annu Rev Pharmacol 14:239–248
Ozols RF, Cunnion RE, Klecker Jr RW, Hamilton TC, Ostchega Y, Parrillo JE, Young RC (1987) Verapamil and Adriamycin in the treatment of drug-resistant ovarian cancer patients. J Clin Oncol 5:641–647, 1987
Pardridge WM, Fierer G (1985) Blood-brain barrier transport of butanol and water relative to N-isopropil-P-iodoamphetamine as the internal reference. J Cereb Blood Flow Metab 5:275–281
Pardridge WM, Landaw EM, Miller LP, Braun LD, Oldendorf WH (1985) Carotid artery injection technique: bounds for bolus mixing by plasma and by brain. J Cereb Blood Flow Metab 5:576–583
Pennock GD, Dalton WS, Roeske WR, Appleton CP, Mosley K, Plezia P, Miller TP, Salmon SE (1991) Systemic toxic effects associated with high-dose verapamil infusion and chemotherapy administration. J Natl Cancer Inst 83:105–110
Riordan JR, Deuchars K, Kartner N, Alon N, Trent J, Ling V (1985) Amplification of P-glycoprotein genes in multidrug-resistant mammalian cell lines. Nature 316:817–819
Sato H, Gottesman MM, Goldstein LJ, Pastan I, Block AM, Sandberg AA, Preisler HD (1990) Expression of the multidrug resistance gene in myeloid leukemias. Leuk Res 14:11–21
Sekido S, Ninomiya K, Iwasaki M (1979) Biologic activity of MCNU: a new antitumor agent. Cancer Treat Rep 63:961–970
Sugawara I, Kataoka I, Morishita Y, Hamada H, Tsuruo T, Itoyama S, Mori S (1988) Tissue distribution of P-glycoprotein encoded by a multidrug-resistant gene as revealed by a monoclonal antibody, MRK16. Cancer Res 48:1926–1929
Terasaki T, Pardridge WM, Denson DD (1986) Differential effect of plasma protein binding of bupivacaine on its in vivo transfer into the brain and salivary gland of rats. J Pharmacol Exp Ther 239:724–729
Thiebaut F, Tsuruo T, Hamada H, Gottesman MM, Pastan I, Willingham MC (1987) Cellular localization of the multidrug-resistance gene P-glycoprotein in normal human tissues. Proc Natl Acad Sci USA 84:7735–7738
Thiebaut F, Tsuruo T, Hamada H, Gottesman MM, Pastan I, Willingham MC (1989) Immunohistochemical localization in normal tissues of different epitopes in the multidrug transport protein 170: evidence for localization in brain capillaries and crossreactivity of one antibody with a muscle protein. J Histochem Cytochem 37:159–164
Tsuji A, Terasaki T, Takanosu K, Tamai I, Nakashima E (1986) Uptake of benzylpenicillin, cefpiramide and cefazolin by freshly prepared rat hepatocytes: evidence for a carrier-mediated transport system. Biochem Pharmacol 35:151–158
Tsuji A, Terasaki T, Takabatake Y, Tenda Y, Tamai I, Yamashima T, Moritani S, Tsuruo T, Yamashita J (1992) P-glycoprotein as the drug efflux pump in primary cultured bovine brain capillary endothelial cells. Life Sci 51:1427–1437
Vick NA, Khandekar JD, Bigner DD (1977) Chemotherapy of brain tumors. The blood-brain barrier is not a factor. Arch Neurol 34:523–526
Yamada K, Ushio Y, Hayakawa T, Kato A, Yamada N, Mogami H (1982) Quantitative autoradiographic measurements of bloodbrain barrier permeability in the rat glioma model. J Neurosurg 57:394–398
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Yamashima, T., Ohnishi, T., Nakajima, Y. et al. Uptake of drugs and expression of P-glycoprotein in the rat 9L glioma. Exp Brain Res 95, 41–50 (1993). https://doi.org/10.1007/BF00229652
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DOI: https://doi.org/10.1007/BF00229652