gms | German Medical Science

58. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie e. V. (DGNC)

Deutsche Gesellschaft für Neurochirurgie (DGNC) e. V.

26. bis 29.04.2007, Leipzig

Development of a screening procedure for the investigation of receptors on human gliomas: Human glioblastoma cells express functional cell membrane receptors for usually applied medical drugs

Entwicklung eines Screening-Verfahrens für die Erkennung von Rezeptoren auf humanen Gliomen: Humane Glioblastomzellen exprimieren funktionelle Zellmembran-Rezeptoren für Medikamente des täglichen Lebens

Meeting Abstract

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  • corresponding author S.A. Kuhn - Klinik für Neurochirurgie, Klinikum der Friedrich-Schiller-Universität, Jena
  • I. Schoenwald - Klinik für Neurochirurgie, Klinikum der Friedrich-Schiller-Universität, Jena
  • R. Kalff - Klinik für Neurochirurgie, Klinikum der Friedrich-Schiller-Universität, Jena
  • R. Reichart - Klinik für Neurochirurgie, Klinikum der Friedrich-Schiller-Universität, Jena

Deutsche Gesellschaft für Neurochirurgie. 58. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie e.V. (DGNC). Leipzig, 26.-29.04.2007. Düsseldorf: German Medical Science GMS Publishing House; 2007. DocP 081

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Veröffentlicht: 11. April 2007

© 2007 Kuhn et al.
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Objective: We addressed the question whether medical drugs applied in daily life could have any effects on human glioma cells, thereby influencing glioma biology. This implied the development of an effective screening procedure.

Methods: We used human glioblastoma cell lines in typical permanent cell culture with application of DMEM supplemented with 10% fetal calf serum, 2mM L-glutamine, 100units/ml penicilline, and 100µg/ml streptomycin. The cells were seeded on glass coverslips for calcium imaging at a density of 5x104 cells/ml. Glioma cells were loaded with 5µM of the calcium indicator Fura-2 in the presence of 0.02% Pluronic. For measurement of calcium changes, cells were illuminated at two alternating wavelengths of 340±5nm and 380±5nm, respectively. Excitation was carried out by employing a monochromator. The emitted light was collected at 530±10nm by a long-term exposure CCD camera QuantiCam coupled to a Zeiss Axioskop equipped with a 20x water immersion objective. The monochromator and CCD camera were controlled by Till Photonics® software.

Results: We developed a method of calcium imaging as a means for receptor screening in gliomas. The imaging workbench was adapted to our aims by addition of mechanical components. Several software programs were written to perform specified experimental settings. In subsequent experiments with histamine, substance P, vasoactive intenstinal polypeptide, neurokinin A, neurokinin B, atropine, dimetinden, metoclopramid, adrenalin, metamizole, clonidine, uradipil, phenytoin, haloperidol, promethazine, cafedrine, theodrenalin, thiamazol, droperidol and physostigmin, a rise of intracellular free calcium ions was observed starting with an abrupt rise up to the top of free calcium ions with an expanded calcium shoulder and a return to base value some seconds after substance wash-out. These changes could be reproduced. Only the buffer controls that should be negative and the application of dexamethason, hydrocortisone and diazepam didn’t exhibit any effect on the intracellular calcium concentration.

Conclusions: Human glioblastoma cells exhibit a rise of intracellular calcium after application of medical drugs, being mediated by functional receptors on the cellular surface, as was confirmed by negative controls. In conclusion, the results show a huge amount of cellular receptors on glioma cells, which can be responsible for further biological effects.