Skip to main content
Log in

Adriamycin binding assay: a valuable chemosensitivity test in human osteosarcoma

  • Origial Papers
  • Clinical Oncology
  • Published:
Journal of Cancer Research and Clinical Oncology Aims and scope Submit manuscript

Summary

The reliability of a simple method evaluating the pattern of subcellular binding of Adriamycin (Adriamycin binding assay, ABA) as an index of sensitivity was demonstrated in different primary cultures and in sensitive and resistant cell lines of human osteosarcoma. After exposure to Adriamycin (10 μg/ml for 30 min at 37°C), living sensitive cells showed selective intranuclear uptake of the drug, whereas in resistant cells no distinct subcellular distribution was observed. The binding pattern of Adriamycin in sensitive and in highly resistant cells was inversely related to the expression of P-glycoprotein. However, low levels of resistance in vitro, not detectable by increased levels of expression of P-glycoprotein, were revealed by ABA. The use of ABA in combination with the estimate of P-glycoprotein expression is recommended in clinical practice as an accurate means for predicting the sensitivity of osteosarcoma to Adriamycin.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Ashihara T (1985) Quantitative cytochemistry: microscope-based cytofluorometry and its application to the analysis of human cancer cells. Acta Histochem Cytochem 17:681–682

    Google Scholar 

  • Baas F, Jongsma APM, Broxterman HJ, Arceci RJ, Housman GL, Scheffer GL, Riethorst A, van Groenigen M, Nieuwint AWM, Joenje H (1990) Non-p-glycoprotein mediated mechanism for multidrug resistance precedes p-glycoprotein expression during in vitro selection for doxorubicin resistance in a human lung cancer cell line. Cancer Res 50:5392–5398

    Google Scholar 

  • Bacci G, Picci P, Ruggieri P, Mercuri M, Avella M, Capanna R, Brach Del Prever A, Mancini A, Gherlinzoni F, Padovani G, Leonessa C, Biagini R, Ferraro A, Ferruzzi A, Cazzola A, Manfrini M, Campanacci M (1990) Primary chemotherapy and delayed surgery (neoadjuvant chemotherapy) for osteosarcoma of the extremities. Cancer 65:2539–2553

    Google Scholar 

  • Bachur NR, Cradock JC (1970) Daunomycin metabolism in rat tissue slices. J Pharmacol Exp Ther 175:331–337

    Google Scholar 

  • Chauffert B, Martin F, Caignard A, Jeannin JF, Leclerc A (1984) Cytofluorescence localization of Adriamycin in resistant colon cancer cells. Cancer Chemother Pharmacol 13:14–18

    Google Scholar 

  • Dolbeare F, Gratzner H, Pallavicini MG, Gray JW (1983) Flow cytometric measurement of total DNA content and incorporated bromodeoxyuridine. Proc Natl Acad Sci USA 80:5573–5577

    Google Scholar 

  • Egorin MJ, Hildebrand RC, Cimino EF, Bachur NR (1974) Cytofluorescence localization of Adriamycin and daunorubicin. Cancer Res 34:2243–2245

    Google Scholar 

  • Fojo A, Akiyama SI, Gottesman MM, Pastan I (1985) Reduced drug accumulation in multiple drug resistant human KB carcinoma cell lines. Cancer Res 45:3002–3007

    Google Scholar 

  • Ganapathi R, Reiter W, Krishan A (1982) Intracellular Adriamycin levels and cytotoxicity in Adriamycin-sensitive and Adriamycin-resistant P388 mouse leukemia cells J Natl Cancer Inst 68:1027–1032

    Google Scholar 

  • Gervasoni JE, Fields SZ, Krishna S, Baker MA, Rosado M, Thuraisamy K, Hindenburg AA, Taub RN (1991) Subcellular distribution of daunorubicin in p-glycoprotein-positive and-negative drug-resistant cell lines using laser-assisted confocal microscopy. Cancer Res 51:4955–4963

    Google Scholar 

  • Gigli M, Rasoanaivo TWD, Millot J-M, Jeannesson P, Rizzo V, Jardillier J-C, Arcamone F, Manfait M (1989) Correlation between growth inhibition and intranuclear doxorubicin and 4-deoxy-4-iododoxorubicin quantitated in living K562 cells by microspectrofluorometry. Cancer Res 49:560–564

    Google Scholar 

  • Goldie JH, Coldman AJ (1984) The genetic origin of drug resistance in neoplasms: implications for systemic therapy. Cancer Res 44:3643–3653

    Google Scholar 

  • Hindenburg AA, Gervasoni JE, Krishna S, Stewart VJ, Rosado M, Lutzky J, Bhalla K, Baker MA, Taub RN (1989) Intracellular distribution and pharmacokinetics of daunorubicin in anthracyline-sensitive and-resistant HL-60 cells. Cancer Res 49:4607–4614

    Google Scholar 

  • Juliano RL, Ling V (1976) A surface glycoprotein modulating drug permeability in chinese hamster ovary cell mutants. Biochim Biophys Acta 455:152–162

    Google Scholar 

  • Kartner N, Riordan JR, Ling V (1983) Cell surface p-glycoprotein is associated with multidrug resistance in mammalian cell lines. Science 221:1285–1288

    Google Scholar 

  • Kessel D, Corbett T (1985) Correlations between anthracycline resistance, drug accumulation and membrane glycoprotein patterns in solid tumors of mice. Cancer Lett 28:187–193

    Google Scholar 

  • Krishan A, Ganapathi R (1980) Laser flow cytometric studies on the intracellular fluorescence of anthracyclines. Cancer Res 40: 3895–3900

    Google Scholar 

  • Kusuzaki K, Litwak GI, Gebhardt MC, Springfield DS, Mankin HJ (1989) Assessment of intracellular Adriamycin (Adr) binding in bone and soft-tissue tumor cells by fluorescence miroscopy. Trans Orthop Res Soc 14:549

    Google Scholar 

  • Luk CK, Tannock IF (1989) Flow cytometric analysis of doxorubicin accumulation in cells from human and rodent cell lines. J Natl Cancer Inst 81:55–59

    Google Scholar 

  • Marsh W, Sickeri D, Center MS (1986) Isolation and characterization of Adriamycin-resistant HL-60 cells which are not defective in the initial intracellular accumulation of drug. Cancer Res 46:187–193

    Google Scholar 

  • Myers CE (1981) Antitumor antibiotics: I. Anthracycline. In: Pinedo HM (ed) Cancer chemotherapy 1980. Excerpta Medica. Amsterdam, pp 66–93

    Google Scholar 

  • Riordan JR, Ling V (1985) Genetic and biochemical characterization of multi-drug resistance. Pharmacol Ther 28:51–75

    Google Scholar 

  • Rotman B, Papermaster BW (1966) Membrane properties of living cells as studied by enzymatic hydrolysis of fluorigenic esters. Proc Natl Acad Sci USA 55:134–141

    Google Scholar 

  • Samuels BL, Murray JL, Cohen MB, Safa AR, Sinha BK, Townsend AJ, Beckett MA, Weichelsbaum RR (1991) Increased glutathione peroxidase activity in a human sarcoma cell line with inherent doxorubicin resistance. Cancer Res 51:521–527

    Google Scholar 

  • Scheper RJ, Bulte JWM, Brakkee JGP, Quak JJ, Schoot E, Balm AJM, Meijer C, Broxterman H, Kuiper CM, Lankelma J, Pinedo HM (1988) Monoclonal antibody (JSB-1) detects a highly conserved epitope on the p-glycoprotein associated with multidrug-resistance. Int J Cancer 42:389–394

    Google Scholar 

  • Serra M, Scotlandi K, Olivari S, Manara MC, Baldini N (1991) Characterization of Adriamycin-resistant variants of two human osteosarcoma cell lines. Eur J Cancer 27 [Suppl 3]:S44

    Google Scholar 

  • Silvestrini R, Gambarucci C, Dasdia T (1970) In vitro biological activity of Adriamycin. Tumori 56:137–148

    Google Scholar 

  • Toffoli G, Viel A, Tumiotto L, Biscontin G, Rossi C, Boiocchi M (1991) Pleiotropic-resistant phenotype is a multifactorial phenomenon in human colon carcinoma cell lines. Br J Cancer 63:51–56

    Google Scholar 

  • Weaver JL, Pine PS, Aszalos A, Schoenlein PV, Currier SJ, Padmanabhan R, Gottesman MM (1991) Laser scanning and confocal microscopy of daunorubicin, doxorubicin, and rhodamine 123 in multidrug-resistance cells. Exp Cell Res 196:323–329

    Google Scholar 

  • Willingham MC, Cornwell MM, Cardarelli CO, Gottesmann MM, Pastan I (1986) Single cell analysis of daunomycin uptake and efflux in multidrug-resistant and-sensitive KB cells: effects of verapamil and other drugs. Cancer Res 46:5941–5946

    Google Scholar 

  • Winkler K, Beron G, Delling G, Heise U, Kabisch H, Purfüst C, Berger J, Ritter J, Jürgens H, Gerein V, Graf N, Russe W, Gruemayer ER, Ertelt W, Kotz R, Preusser P, Prindull, G, Brandeis W, Landbeck G (1988) Neoadjuvant chemotherapy of osteosarcoma: results of a randomized cooperative trial (COSS-82) with salvage chemotherapy based on histological tumor response. J Clin Oncol 6:329–337

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

This study was supported by a grant from the Associazione Italiana per la Ricerca sul Cancro (A.I.R.C.).

Rights and permissions

Reprints and permissions

About this article

Cite this article

Baldini, N., Scotlandi, K., Serra, M. et al. Adriamycin binding assay: a valuable chemosensitivity test in human osteosarcoma. J Cancer Res Clin Oncol 119, 121–126 (1992). https://doi.org/10.1007/BF01209668

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01209668

Key words

Navigation