Dose-related effects of psoralen and ultraviolet light on the cell cycle of murine melanoma cells.

Cloudman (S91) murine melanoma cells were treated with 4'-hydroxymethyltrioxsalen (HMT), a bifunctional psoralen and exposed to long-wavelength (365 nm) ultraviolet light. DNA content of the cells stained with propidium iodide was measured by flow cytometry, and cell cycle phases were delineated from the DNA histograms by using a curve-fitting routine. We found that HMT in combination with long-wavelength (365 nm) ultraviolet irradiation blocked melanoma cells in different phases of the cell cycle, depending on the dose of long-wavelength (365 nm) ultraviolet light and the concentration of HMT. The binding of [3H]HMT to DNA was measured parallel with cell cycle analyses. Treatments with HMT at concentrations corresponding to about 1 HMT bound per 10(6) base pairs of DNA led to the accumulation of cells with predominantly G2 DNA content. At higher concentrations (2 to 3 HMT/10(6) base pairs), the cells were blocked in the S and G1 phases. In conclusion, we have shown that extremely sparse substitution of HMT to DNA blocks melanoma cells in the G2 phase or other phases of the cell cycle in a dose-dependent manner.