title: Characterization of peripheral-type benzodiazepine binding sites on human lymphocytes and lymphoma cell lines and their role in cell growth.
abstract: Peripheral-type benzodiazepine binding sites (PBRs) are ubiquitous in mammalian tissues. However, the physiological role of PBRs has not yet been clarified. In this study we characterized a saturable and high affinity binding site for [3H]Pk 11195 (isoquinoline carboxamide derivative) on human lymphocytes and different lymphoma cell lines. Binding parameters of the human T-lymphoma cell line CCRF-CEM came closest to values for lymphocyte binding. Thus, the CCRF-CEM cell line appears to be a suitable lymphocyte cell model for further study of PBRs. To evaluate the pharmacological specificity of binding to human lymphocytes and CCRF-CEM cells we investigated the potency of different ligands to displace [3H]Pk 11195 from its binding site. Pk 11195 was found to be the most potent inhibitor followed by 4'-chlorodiazepan (Ro5-4864) and diazepam (range of inhibition constants from 6.7 x 10(-9) M to 3.6 x 10(-7) M), whereas ligands specific for the central-type receptor like clonazepam and flumazenil had no displacing potency in the tested concentration range (10(-10)-10(-4) M). Since it was assumed that PBRs might be involved in the regulation of cell growth and differentiation, we studied the influence of PBR ligands on cell growth and survival using a quantitative colorimetric assay (MTT). Ligands which bind selectively to PBRs inhibited cell multiplication in vitro. However, half-effective concentrations (EC50) were in the micromolar range and above therapeutic in vivo concentrations (range of EC50 values from 2.4 x 10(-5) M to 1.5 x 10(-4) M). Clonazepam and flumazenil had no inhibiting potency in the tested concentration range (10(-10)-10(-4) M). Although the difference between values for displacing potency and ability to inhibit cell multiplication cannot be explained as yet, it is interesting that all PBR-ligands followed the same sequence in displacing [3H]Pk 11195 and inhibiting cell multiplication and that central type ligands were ineffective in both assays. This association suggest a mediating role of PBR binding in cell growth.
authors.affiliation: Dr Margarete Fischer-Bosch-Institute of Clinical Pharmacology, Stuttgart, Federal Republic of Germany.
authors.forename: B E
authors.initials: BE
authors.lastname: Alexander
authors.forename: E
authors.initials: E
authors.lastname: Roller
authors.forename: U
authors.initials: U
authors.lastname: Klotz
mesh_terms: D000328:Adult; D001665:Binding Sites; D002455:Cell Division; D002460:Cell Line / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D005260:Female; D006801:Humans; D007546:Isoquinolines / Q000493:pharmacokinetics; D008214:Lymphocytes / Q000378:metabolism*; D008223:Lymphoma / Q000378:metabolism*; D008297:Male; D011963:Receptors, GABA-A / Q000187:drug effects / Q000378:metabolism*; D014316:Tritium
chemical_list: D007546:Isoquinolines; D011963:Receptors, GABA-A; D014316:Tritium; C037850:PK 11195