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Intracellular localization of drugs in cultured tumor cells by ion microscopy and image processing
J P Berry1, G Lespinats, F Escaig
1SC 27 de l'INSERM, Faculté de Médicine, Créteil, France.
Abstract:
Several drugs, containing a halogen atom, F or Br, that are being used in antiviral or anticancer therapy, were studied for their localization in cultured cells by ion microanalysis. The association allows to reduce the exposure time to define the intracellular localization of the studied element. The topography of the cells is given by the image of the polyatomic ion 26CN-. The image of the distribution of 81Br- or 19F-, coded in another color scale, can be superimposed, giving a polychromic image of the cell, thus showing the intracellular localization of the compound. MCF-7 tumor cells were cultured in the presence of pyrimidine derivatives. 5-Bromo-2'-deoxyuridine (BUdR) and 5-trifluorothymidine (F3TdR) were localized in the nucleus, 5-fluoro-2'-deoxyuridine (FUdR) in the nucleus and only in some nucleoli. The method is simple and rapid, as compared with techniques using radiolabeled compounds, or with immunocytochemical techniques. It is possible to observe two different compounds in the same cell. It could be applied to other compounds containing a halogen atom.
Insights
This study introduces ion microanalysis to rapidly visualize halogenated drugs (like F or Br) within cultured cells. The technique effectively maps drug localization in cell nuclei and nucleoli, aiding antiviral and anticancer drug development.
Area of Science:
- Cell Biology
- Pharmacology
- Analytical Chemistry
Background:
- Halogenated drugs containing fluorine (F) or bromine (Br) are crucial in antiviral and anticancer therapies.
- Understanding the intracellular localization of these drugs is vital for optimizing their efficacy and minimizing side effects.
- Existing methods for drug localization can be time-consuming or require specialized labeling.
Purpose of the Study:
- To develop and validate a rapid ion microanalysis technique for determining the intracellular localization of halogenated drugs.
- To assess the cellular distribution of specific pyrimidine derivatives (BUdR, F3TdR, FUdR) in MCF-7 tumor cells.
- To demonstrate the potential of ion microanalysis for simultaneous visualization of multiple halogenated compounds within cells.
Main Methods:
- Utilized ion microanalysis to detect and map the distribution of 81Br- and 19F- ions in cultured MCF-7 cells.
- Employed 26CN- ion imaging for cell topography and superimposed elemental distribution maps for polychromic visualization.
- Cultured MCF-7 cells in the presence of pyrimidine derivatives: 5-Bromo-2'-deoxyuridine (BUdR), 5-trifluorothymidine (F3TdR), and 5-fluoro-2'-deoxyuridine (FUdR).
Main Results:
- Successfully localized BUdR and F3TdR within the nucleus of MCF-7 cells.
- Determined that FUdR localized to the nucleus and specifically within certain nucleoli.
- Demonstrated that ion microanalysis significantly reduces exposure time compared to radiolabeled or immunocytochemical methods.
- Showcased the ability to visualize two different halogenated compounds within the same cell simultaneously.
Conclusions:
- Ion microanalysis provides a simple, rapid, and effective method for determining the intracellular localization of halogenated drugs.
- The technique offers advantages in speed and simplicity over traditional methods, facilitating drug discovery and development.
- This approach is applicable to a wide range of compounds containing halogen atoms for cellular localization studies.