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Published on: March 30, 2014
A fluorescence-based assay for monitoring clinical drug resistance
Ariane Deniset-Besseau1, François-Alexandre Miannay, Corinne Laplace-Builhé
1Institut des Sciences Moléculaires d'Orsay, Université Paris-Sud, Orsay, France. ariane.deniset@u-psud.fr
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.
Insights
This study introduces a new assay to predict chemotherapy response by analyzing drug fluorescence in cancer cells. The assay is simple, reliable, and inexpensive for monitoring multidrug resistance (MDR).
Area of Science:
- Oncology
- Cell Biology
- Medical Diagnostics
Background:
- Multidrug resistance (MDR) significantly hinders cancer treatment efficacy.
- MDR involves altered drug transport across cancer cell membranes.
- Predicting patient response to chemotherapy is crucial for effective treatment.
Purpose of the Study:
- To develop and validate a novel assay for monitoring patient responses to chemotherapy.
- To assess the utility of the assay in predicting efficacy of doxorubicin and MVAC therapy.
Main Methods:
- Utilized a slide-based cell imaging technique leveraging the fluorescent properties of doxorubicin.
- Optimized the assay using breast and urothelial cancer cell lines.
- Applied the assay to clinical specimens including fine needle breast aspiration biopsy and urine cytology.
Main Results:
- Observed identical drug fluorescence behavior in clinical specimens and cultured cells.
- Sensitive cells showed nuclear doxorubicin localization, while resistant cells exhibited cytoplasmic localization.
- Fluorescence intensity differences were quantifiable through spectral analysis.
Conclusions:
- The developed assay provides valuable clinical information for predicting patient response to doxorubicin and MVAC therapy.
- The assay is technically simple, reliable, and cost-effective.
- Effective on both confocal and standard epifluorescence microscopes.
