Fotemustine (FM) is a member of the chloroethylnitrosourea class of alkylating agents that has been proven active against the disseminated melanoma and primary brain tumours . Spontaneous decomposition of nitrosoureas generates electrophilic species, which are responsible for DNA alkylation, thus producing therapeutic effects. The generation of isocyanates cause toxic side effect
of FM which are monitored through SB273005 carbamoilation of proteins . The monofunctional alkylating agent dacarbazine (DTIC) is approved and frequently used for the treatment of melanoma. Relative response after DTIC treatment is observed in 15 to 20% of cases with short duration [5, 6]. Due to the inherent drug-resistant characteristic of this disease, chemotherapy
is an ineffective mean of treating see more malignant melanoma. The reasons for the chemoresistant phenotype in human melanoma are not well understood and are probably multifactorial. Some forms of specially localized melanoma tumors, are presently treated with therapeutic proton beams giving positive results . Physical properties of protons, 4SC-202 such as their well defined range, with the small lateral scattering and high energy deposition within the Bragg peak maximum, made this type of therapy suitable for localized melanomas. In order to treat the malignant growth with protons
so that the desired uniform dose can be delivered over the large volume at the given depth, the Bragg peak is spread out by the modulation of proton energy, followed by the slight increase of the entrance dose. Various authors have reported data on modulated proton beams with energy less than 100 MeV which are used for the treatment of eye melanoma [8, 9]. With the goal to find a more efficient way to treat melanoma, combined treatments of either oxyclozanide FM or DTIC with proton irradiations were examined. In our previous studies, we investigated the effects of proton irradiations and single drug treatments on HTB140 cells, as well as the effects of proton irradiations on these cells that were pre-treated with FM or DTIC [10–12]. The objective of the present study is to examine whether the change in order and duration of treatments applied have the influence on cell inactivation level. Therefore, cell viability, proliferation, survival and cell cycle distribution were investigated on HTB140 human melanoma cells that were first irradiated and than exposed to FM or DTIC. Methods Cell Culture The human melanoma HTB140 cells were purchased from the American Tissue Culture Collection (Rockville, MD, USA). They were grown in the RPMI1640 medium supplemented with 10% fetal bovine serum, penicillin-streptomycin and L-glutamine.