Influence of irradiation on the metabolism of melanoma cells and metastasis in mice.

Irradiation is a powerful tool for the therapy of solid tumors. But often single cells elude this treatment and constitute a basis for recurrence of the primary tumor and formation of metastases. Until today it is unclear which properties enable some cells to this. One possible explanation could be predicted on irradiation-dependent metabolic changes which lead to a predisposition of certain cells to show enhanced survival and migratory activity. The aim of this study was to investigate metabolic properties and proliferation of irradiated melanoma cells in vitro and their ability to form metastases in vivo.
We applied different single-dose X-ray irradiation (200kV X-rays, 0.5mm Cu, ~ 1.2 Gy min-1; 1, 2, 5, 7, 10, and 20 Gy) to murine B16-F10 melanoma cells. At particular times we analyzed cell viability, growth properties and cell cycle distribution. Furthermore, we analyzed the cellular uptake of the radiotracers 2-[18F]fluoro-2-deoxy-D-glucose ([18F]FDG) and 3-O-methyl-[18F]fluoro-L-DOPA ([18F]OMFD), providing information about the glucose and amino acid metabolism before and after irradiation. Additionally, we performed in vivo studies in a syngeneic mouse model to analyze the capability of irradiated melanoma cells to form lung metastases after injection into the tail vein of NMRI mice.
In a dose-dependent manner we detected a decrease in cell viability and cell growth properties starting 3 days after irradiation. Decreased cell growth persists up to 1 week for 5 Gy irradiated cells and up to 2 weeks for 10 Gy irradiated cells. After this periods growth of irradiated cells is comparable to control cells. Cell cycle analyses showed an increase in G2/M phase cells up to 3 days after X-ray followed by an increase in S phase cells 6 days after X-ray. At this point of time uptake of radiotracers was altered inasmuch as [18F]FDG uptake decreased, whereas [18F]OMFD uptake increased. Our in vivo studies showed a loss of lung metastases when cells were irradiated (10 Gy) before injection. If irradiated cells were allowed to recover for 2 weeks before injection, mice again developed lung metastases although to a lesser extent than control mice.
We conclude that irradiation of melanoma cells leads to a dose-dependent decrease in cell viability, growth properties and glucose uptake. Cell cycle analyses suggest an arrest in the G2/M phase. One week after irradiation compensating mechanisms of these effects seems to start as indicated by the uptake of [18F]OMFD, the increase in S phase cells and recovered growth of low-dose (5 Gy) irradiated cells. Two weeks after irradiation cell growth is completely recovered in vitro. Accordingly, in vivo studies reveal that irradiated melanoma cells are able to resume their metastatic potential within two weeks, even though to a lesser extent than before irradiation. The questions why and how some cells modulate their metabolism and thus re-start proliferation and why metastasis is influenced in vivo although growth properties are recovered in vitro, need to be further investigated.