ABSTRACT
Hydrogen peroxide (H2O2) and free radicals cause oxidative stress, which induces cellular injuries, metabolic dysfunction, and even cell death in various clinical abnormalities. Fullerene (C60) is critical for scavenging oxygen free radicals originated from cell metabolism, and reduced glutathione (GSH) is another important endogenous antioxidant. In this study, a novel water-soluble reduced glutathione fullerene derivative (C60-GSH) was successfully synthesized, and its beneficial roles in protecting against H2O2-induced oxidative stress and apoptosis in cultured HEK 293T cells were investigated. Fourier Transform infrared spectroscopy and (1)H nuclear magnetic resonance were used to confirm the chemical structure of C60-GSH. Our results demonstrated that C60-GSH prevented the reactive oxygen species (ROS)-mediated cell damage. Additionally, C60-GSH pretreatment significantly attenuated H2O2-induced superoxide dismutase (SOD) consumption and malondialdehyde (MDA) elevation. Furthermore, C60-GSH inhibited intracellular calcium mobilization, and subsequent cell apoptosis via bcl-2/bax-caspase-3 signaling pathway induced by H2O2 stimulation in HEK 293T cells. Importantly, these protective effects of C60-GSH were superior to those of GSH. In conclusion, these results suggested that C60-GSH has potential to protect against H2O2-induced cell apoptosis by scavenging free radicals and maintaining intracellular calcium homeostasis without evident toxicity.
Subject(s)
Humans , Antioxidants , Pharmacology , Apoptosis , Calcium , Metabolism , Caspase 3 , Genetics , Metabolism , Cell Survival , Fullerenes , Chemistry , Pharmacology , Gene Expression Regulation , Glutathione , Pharmacology , HEK293 Cells , Hydrogen Peroxide , Pharmacology , Ion Transport , Malondialdehyde , Metabolism , Oxidative Stress , Proto-Oncogene Proteins c-bcl-2 , Genetics , Metabolism , Reactive Oxygen Species , Metabolism , Signal Transduction , Superoxide Dismutase , Metabolism , bcl-2-Associated X Protein , Genetics , MetabolismABSTRACT
Human cancer cell lines express human choriogonadotropin [hCG], its subunits and derivatives, regardless of their origin and type. It appears that hCG is a common phenotype in human cancer cell lines. In this research, the effects of hCG targeting monoclonal antibodies [7D9, T18H7 and T8B12] on human cancer cell lines were evaluated. Monoclonal antibody secreting hybridomas were proliferated and injected intraperitoneally to Balb/C mice after treatment with pristine. Two weeks later, ascites fluid was collected. Purification of aforementioned antibodies from ascites fluid was performed using G-protein affinity followed by ion exchange chromatography. SDS-PAGE and ELISA confirmed the structure and functional integrity of the purified antibodies, respectively. Two human cancer cell lines "Hela" and "MDA" were treated by the purified antibodies. Three days later, different wells were imaged and the cells counted. SDS-PAGE gel [None-reducing] indicated consistency of band migration patterns with control antibodies. ELISA test using hCG antigens indicated that the produced antibodies could detect hCG antigens. Cell lines were cultured and treated with different concentrations of each antibody. Counting and imaging different wells of treated plates, indicated that 7D9 antibody had a more significant [P<0.01] cytotoxic effect on cancer cell lines than the control cells. HCG targeting monoclonal antibodies can be used for targeted cancer therapy, as human cancer cells express hCG gene. 7D9 antibody that exhibits protease activity is a proper candidate for this purpose, as it possesses both antagonistic and enzymatic properties
Subject(s)
Humans , Animals, Laboratory , Chorionic Gonadotropin , Cell Line, Tumor , Mice, Inbred BALB C , Fullerenes , Ascitic FluidABSTRACT
<p><b>AIM</b>To evaluate the effect of fullerenol on the antioxidant system of goat epididymal sperm.</p><p><b>METHODS</b>Fresh epididymides of adult goats were obtained from local slaughter houses and sperm were collected by chopping the epididymis in modified Ringer's phosphate solution (RPS medium). After several washings the sperm samples were equally dispersed in RPS medium and incubated with fullerenol (1, 10 and 100 micromol) and FeSO(4)/ascorbate (40/200 micromol) with or without fullerenol (1, 10 and 100 micromol) for 3 h at 32 degree C. After incubation, an aliquot of sperm samples were homogenized and centrifuged and the supernatant used for biochemical studies.</p><p><b>RESULTS</b>In FeSO(4)/ascorbate-incubated samples, the activities of antioxidant enzymes, superoxide dismutase, glutathione peroxidase and glutathione reductase, were decreased while lipid peroxidation increased as compared to the control sperm samples. In fullerenol-incubated sperm samples, the activities of superoxide dismutase, glutathione peroxidase and glutathione reductase were increased while lipid peroxidation was decreased in a dose-dependent manner. Co-incubation of sperm with fullerenol (1,10 and 100 micromol) and FeSO(4)/ascorbate (40/200 micromol) increased the activities of antioxidant enzymes and prevented the iron-induced elevation of lipid peroxidation in a dose-dependent manner.</p><p><b>CONCLUSION</b>Fullerenol reduces iron-induced oxidative stress in epididymal sperm of goat by increasing the activities of antioxidant enzymes and decreasing lipid peroxidation.</p>