The Influence Of The ERK Signaling Pathway By Regulated Of Intracellular Reactive Oxygen Species

Cu/Zn superoxide dismutase?SOD1?,one of the most important and widely distributed antioxidant intracellular enzymes,catalyzed the dismutation of superoxide to peroxide hydrogen.SOD1 as the cancer potential therapeutic targets,many anticancer drugs designed are based on the inhibition of SOD1.Our team based on bis?thiosemicarbazone?complexes?ATSM?,which have a high affinity for copper,synthesized a series of copper cHeLating agents?LD series?,some has inhibitory effect on SOD1.The inhibitors can effectively inhibit the activity of intracellular SOD1 and decreased the intracellular H₂O₂ levels significantly.The main results including:1.We study the toxicity of LD series inhibitors for HeLa cell by MTT assay.The results show that these compounds have low toxicity.The cytotoxicity of LD25,LD34,LD94,LD100 are larger,the most suitable concentration is 50 mM;The cytotoxicity of LD18,LD27,LD29,LD32 is lower,the largest concentration of up to 100 mM.2.We detect the across membrane function of LD94 and LD100 as representative of cHeLating agents with Inverted fluorescence microscope.The results show that these inhibitors can easily through the cell membrane and stay in the cytoplasm because of small molecules.3.By fluorescent quantitative PCR assays,we detect the influence of intracellular SOD1 gene expression by LD series inhibitors.The results show that these cHeLating agents have no influence of SOD1 gene expression.4.Protein immunoblot experiment results show that several typical SOD1 inhibitors can reduce the level of intracellular phosphorylation of ERK.5.We detect the influence of LD100 on different cell apoptosis by flow cytometry.The results show that LD100 can promote the apoptosis of HeLa cells,and had no effect on normal cell line.In a word,LD series inhibitors have lower cytotoxicity,can easily across the cell membrane and have no influence of SOD1 gene expression.The inhibitors can reduce the level of intracellular phosphorylation of ERK by inhibit the activity of SOD1 and regulate the intracellular ERK signal transduction pathway by the apoptosis mechanism.