Flow cytometric monitoring of glutathione content and anthracycline retention in tumor cells

Swagata Nair, Shivendra V. Singh, Awtar Krishan

Research output: Contribution to journalArticlepeer-review

26 Scopus citations

Abstract

We have used an enzymatic (spectrophotometric) and a flow cytometric (GSH-MBCL) method to compare the glutathione (GSH) content of doxorubicin sensitive (P388) and resistant (P388/R-84) murine leukemic and human lung cancer cells. The flow cytometric analysis revealed that GSH-MBCL conjugate formation was dependent on glutathione-S-transferase (GST) activity. The human solid tumor cell lines exhibited extensive heterogeneity, high GSH content, and GST activity. In contrast to the enzymatic method, the flow cytometric method did not accurately reflect the 95% reduction in GSH content of cells treated for 24 h with 100 μM BSO. Possible reaction of MBCL with other sulfhydryl groups (other than GSH) in BSO-treated cells may be responsible for this discordance. We have also shown the feasibility of using dual parameter flow cytometry to monitor cellular anthracycline (daunorubicin) retention and GSH-MBCL conjugate fluorescence in human tumor cells. These two parameters, which measure drug retention and cellular detoxification, are believed to be the important determinants of chemoresistance in tumor cells.

Original languageEnglish (US)
Pages (from-to)336-342
Number of pages7
JournalCytometry
Volume12
Issue number4
DOIs
StatePublished - 1991

Keywords

  • anthracyclines
  • daunorubicin
  • drug detoxification
  • drug transport
  • glutathione
  • multiple drug resistance

ASJC Scopus subject areas

  • Hematology
  • Cell Biology
  • Pathology and Forensic Medicine
  • Biophysics
  • Endocrinology

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