Cell cycle dependent sensitivity of human melanoma cells to melphalan is correlated with the activity and cellular concentration of glutathione transferases.
Glutathione transferases (GSTs) are enzymes involved in the resistance of tumor cells to bifunctional alkylating cytostatic drugs. We investigated the melphalan sensitivity together with activity and cellular concentration of GST isoenzymes of human melanoma cell line RPMI 8322 in different phases of the cell cycle. By centrifugal elutriation three cell fractions containing different proportions of cells in the G1 phase were isolated. Melphalan sensitivity was estimated by the colony formation assay. The cell fraction with the largest proportion of G1 cells was more sensitive to the drug than the fractions enriched in S and G2 cells. The GST activity of the cell fractions was measured with 1-chloro-2,4-dinitrobenzene (CDNB) as substrate and the concentrations of GST P1-1, GST M1-1 and GST A1-1 were quantitated by use of isoenzyme-specific ELISA. The results show that there were less GST activity and lower GST P1-1 and A1-1 concentrations in the G1 cell enriched fraction, demonstrating a cell cycle dependence of GST expression. Thus, the cell fraction most sensitive to melphalan had the highest proportion of G1 cells and displayed the lowest GST activity, suggesting that the cell cycle dependent sensitivity to melphalan may at least partially depend on the expression of GSTs.
Jungnelius
U
U
Department of General Oncology, Karolinska Hospital, Stockholm, Sweden.
Hao
X Y
XY
Skog
S
S
Castro
V M
VM
Mannervik
B
B
Ringborg
U
U
D002453:Cell Cycle / Q000502:physiology; D002458:Cell Fractionation; D002498:Centrifugation; D004354:Drug Screening Assays, Antitumor; D005982:Glutathione Transferase / Q000378:metabolism*; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism*; D008545:Melanoma / Q000188:drug therapy* / Q000201:enzymology* / Q000473:pathology; D008558:Melphalan / Q000494:pharmacology*; D013347:Subcellular Fractions / Q000201:enzymology; D014407:Tumor Cells, Cultured / Q000187:drug effects
D007527:Isoenzymes; D005982:Glutathione Transferase; D008558:Melphalan