| OBJECTIVE:Synthesis and characterization of disulfiram(DSF)derivatives,the preliminary study on their antitumor activities in vitro and their radioprotective effects in vivo.METHODS:Two derivatives were synthesized in two steps,and their structures were confirmed using 1H NMR,13C NMR and LC-MS.CCK-8 cytotoxicity assay was used to compare the inhibitory effects of DSF and its derivatives on proliferation of MCF-7 and A549 cells with or without FBS.Effects of NAC on anti-tumor activities of these compounds were detected in the same way.Whether DSF or its derivatives could induce apoptosis was analyzed by flow cytometry.Expressions of related apoptotic proteins in MCF-7 and A549 cells were detected by Western blot.The mice were given γ-ray whole body irradiation.The radiation protective effects of DSF and its derivatives were evaluated by the 30-day survival rate experiment of lethal dose radiation,the sublethal dose radiation anti-radiation physiological index of mice and the anti-oxidative test.RESULTS:Results from the spectral analyses showed that the structures of the derivatives were consistent with the expectation.The CCK-8 results showed that the inhibitory effects of the two derivatives on these two kinds of tumor cells were stronger than that of DSF,and there were significant differences on the value of IC50.There was an obvious decline in the proliferation inhibitory effects of the derivatives in the absence of FBS.In addition,the inhibitory effects on the proliferation of MCF-7 and A549 cells was significantly weakened after the addition of NAC(P<0.05).Flow cytometry analyses showed that the apoptosis rates of MCF-7 and A549 cells which were treated with the two derivatives increased compared with DSF(P<0.05 or P<0.01).The results from Western blot analyses showed that the two derivatives increased the expressions of BAX and decreased the expressions of PARP in the MCF-7 and A549 cells.The 30-day survival experiment showed that oral administration of DSF-1 and DSF-2 could separately increase the 30-day survival rates of radiation-injured mice by 50%,and significantly prolong the average survival time of radiation-injured mice by 12 days,with the protection indexes of 2.22.Intraperitoneal injection of DSF-2 could increase the 30-day survival rate of radiation-injured mice by about 60%,prolong the average survival time of radiation-injured mice by 7 days,and the protection index was 1.366.The sublethal dose index experiment showed that compared with the irradiation control mice,oral administration of DSF-1 and DSF-2 significantly increased the thymus indexes of irradiated mice,and protect the hematopoiesis and immune systems of irradiated mice.The difference was statistically significant.Antioxidant tests showed that oral administration of DSF-1 or DSF-2 could increase the content of glutathione(GSH)in the liver of mice.Intraperitoneal injection of DSF-2 could increase the contents of GSH,total superoxide dismutase(T-SOD)and catalase(CAT)and decrease the content of malondialdehyde(MDA)in the livers of mice.CONCLUSION:The two derivatives show anti-tumor activities.In addition,the inhibitory effects on the two kinds of tumor cells in the presence of FBS are better than that of DSF.The mechanisms may be related to the expression of apoptosis-related proteins and the increase of intracellular ROS production.The two compounds have antioxidant effects and protective effects on immune and hematopoietic system caused by ionizing radiation. |
