Measurement of P-glycoprotein expression in human neuroblastoma xenografts using in vitro quantitative autoradiography(284 views) Fonti R, Levchenko A, Mehta BM, Zhang J, Tsuruo T, Larson SM
Nucl Med Biol (ISSN: 0969-8051, 0883-2897), 1999 Jan; 26(1): 35-41.
Affiliations: Department of Nuclear Medicine, Mem. Sloan-Kettering Cancer C., New York, NY, United States
Inst. of Molec. and Cell. Biosci., University of Tokyo, Tokyo, Japan
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Gudkov, A. V., Multiple drug resistance in tumor cells: Manifestations, clinical basis, clinical aspects (1987) Mol. Gen. Microbiol. Virusol., 3, pp. 3-9
Hulme, E. C., (1992) Receptor-Ligand Interactions. a Practical Approach, , New York: Oxford University Press
Juliano, R. L., Ling, V., A surface glycoprotein modulating drug permeability in Chinese hamster ovary cell mutants (1976) Biochim. Biophys. Acta, 455, pp. 152-162
Mehta, B. M., Levchenko, A., Rosa, E., Kim, S. W., Winnick, S., Zhang, J. J., Kalaigian, H., Larson, S. M., Evaluation of carbon-14-colchicine biodistribution with whole body quantitative autoradiography in colchicine sensitive and resistant xenografts (1996) J. Nucl. Med., 37, pp. 312-314
Merkel, D. E., Fuqua, S. A. W., Tandon, A. K., Hill, S. M., Buzdar, A. U., McGuire, W. L., Electrophoretic analysis of 248 clinical breast cancer specimens for P-glycoprotein overexpression or gene amplification (1989) J. Clin. Oncol., 7, pp. 1129-1136
Motulsky, H. J., Ransnas, L. A., Fitting curves to data using nonlinear regression: A practical and nonmathematical review (1987) FASEB J, 1, pp. 365-374
Scott, A. M., Rosa, E., Mehta, B. M., Divgi, C. R., Finn, R. D., Biedler, J. L., Tsuruo, T., Larson, S. M., In vivo imaging and specific targeting of P-glycoprotein expression in multidrug resistant nude mice xenografts with 125I-MRK16 monoclonal antibody (1995) Nucl. Med. Biol., 22, pp. 497-504
Shen, D. W., Pastan, I., Gottesman, M. M., In situ hybridization analysis of acquisition and loss of human multidrug resistance gene (1988) Cancer Res, 48, pp. 4334-4339
Sirotnak, F. M., Yang, C. H., Mines, L. S., Oribe, E., Biedler, J. L., Markedly altered membrane transport and intracellular binding of vincristine in multidrug-resistant Chinese hamster cells selected for resistance to vinca alkaloids (1986) J. Cell Physiol., 126, pp. 266-274
Van Der Bliek, A. M., Borst, P., Multidrug resistance (1989) Adv. Cancer Res., 52, pp. 165-202
Young, A. B., Frey, K. A., Agranoff, B. W., Receptor assay: In vitro and in vivo (1986) Positron Emission Tomography and Autoradiography: Principles and Applications for the Brain and Heart, pp. 73-111. , M. Phelps, J. Mazziotta, & H. Schelbert. New York: Raven Press
Measurement of P-glycoprotein expression in human neuroblastoma xenografts using in vitro quantitative autoradiography
P-glycoprotein (P-gp) has a role in multidrug resistance (MDR) encountered in human cancers. In this study, we used the colchicine-resistant cell line BE(2)-C/CHCb(0.2), a strain of neuroblastoma cell line BE(2)-C, as a model to measure variations of P-gp expression in cells grown in vitro and in vivo. Cells were cultured in the medium supplemented with colchicine. At the beginning of the study the drug was withdrawn and, after 22 days, added back to the culture medium. Cells were harvested at various time points and xenografted in nude mice. P-gp COntent in cells was measured by self- competitive binding assay and in tumors, by quantitative autoradiography (QAR). Both assays were carried out using 125I-labeled monoclonal antibody MRK16, reactive with P-gp. Concentration of P-gp in cells varied from a maximum of 1,361 pmol/g in the presence of colchicine to a minimum of 374 pmol/g in the absence of colchicine in the culture medium. P-gp concentration in the tumors ranged from 929 to 188 pmol/g, which correlated with P-gp content in the cells at the time of their injection in the mice. QAR is an accurate and reliable method to quantify P-gp expression in tumors. Changes in colchicine concentration in the ambient medium of BE(2)-C/CHCb(0.2) cells growing in vitro resulted in a change in phenotype of P-gp expression, which was stable under conditions of in vivo growth over approximately 9 cell divisions in nude mice xenografts. Therefore, P-gp content in xenografts depends only on the level of resistance of the cells at the time of their injection in the mice.
Measurement of P-glycoprotein expression in human neuroblastoma xenografts using in vitro quantitative autoradiography
No results.
Measurement of P-glycoprotein expression in human neuroblastoma xenografts using in vitro quantitative autoradiography