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A fluorescence-based assay for monitoring clinical drug resistance
  1. Ariane Deniset-Besseau1,
  2. François-Alexandre Miannay1,2,
  3. Corinne Laplace-Builhé3,
  4. Philippe Vielh4,
  5. Sandrine Lécart2,
  6. Bashir A Lwaleed5,
  7. Pascal Eschwege6,7,
  8. Marie-Pierre Fontaine-Aupart1,2
  1. 1Institut des Sciences Moléculaires d'Orsay, Université Paris-Sud, Orsay, France
  2. 2Centre de Photonique Biomédicale, Centre Laser de l'Université Paris-Sud, Université Paris-Sud, Orsay, France
  3. 3Imaging and Cytometry Platform, Institut de Cancérologie Gustave Roussy, Villejuif, France
  4. 4Department of Biopathology and Translational Research Laboratory, Institut de Cancérologie Gustave Roussy, Villejuif, France
  5. 5Faculty of Health Sciences, University of Southampton, Southampton General Hospital, Southampton, UK
  6. 6Service d'Urologie, CHU Bradois, Vandoeuvre-Lès-Nancy, France
  7. 7Urology Department, Antony Private Hospital, Antony, France
  1. Correspondence to Dr Ariane Deniset-Besseau, Laboratoire de Chimie-Physique, UMR 8000, Université Paris-Sud, bâtiment 201 porte 2, Orsay 91405, France; ariane.deniset{at}u-psud.fr

Abstract

Background and aims Multidrug resistance (MDR) limits effectiveness in treating malignancy by modifying internalisation and/or externalisation of drugs through cancer cell membranes. In this study we describe an assay to monitor patients’ responses to chemotherapy.

Methods The assay is based on the fluorescent properties of doxorubicin alone as well as in combination with methotrexate, vinblastine, doxorubicin and cisplatin (MVAC). The slide-based cell imaging technique was first optimised using a panel of breast and urothelial cancer cell lines and then extended to fine needle breast aspiration biopsy and urine cytology.

Results The drug fluorescence behaviour observed on smears of clinical specimens is identical to that obtained using fixed cultured cells. The fluorescence of sensitive cells to chemotherapy is mainly localised in the nucleus, whereas resistant cells show a weak fluorescence signal localised in the cytoplasm. The difference in terms of fluorescence intensity is also highlighted through fluorescence spectra.

Conclusions The results suggest that the assay provides clinically valuable information in predicting responses to doxorubicin and/or MVAC therapy. Originally set up on a confocal microscope, the assay was also effective using a standard epifluorescence microscope; as such it is technically simple, reliable and inexpensive.

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