Autophagy And Apoptosis In Hepatocellular Carcinoma Induced By EF25-（GSH）₂:A Novel Curcumin Analog

Curcumin,a spice component as well as a traditional Asian medicine,has been reported to inhibit proliferation of a variety of cancer cells.Curcumin can adjust its target molecule to regulate cell proliferation and apoptosis,these molecules are important in a number of cell transcription factors,growth factors and their associated receptors,cytokines,enzymes and regulatory genes.On this basis,curcumin can inhibit the*proliferation of theoretically almost all types of tumor cells.However,curcumin used in practice but is limited by its relatively low metabolic activity in vivo biological.Further curcumin absorption body which is also constrained by the hydrophobic nature.Further,since the yellow curcumin easy infection,its application in the many skin diseases is also greatly limited..For these reasons,the Institute of Chemistry and Pharmacology,Emory University researchers synthesized by chemical methods,in order to obtain a variety of main core structure of curcumin analogs for the wizard.Through high-throughput screening methods,and found one of the analogs have a high biological activity,and further research.Early results show which EF24,EF25 and EF25-(GSH)2 has a better anti-tumor cell proliferation activity.And this one EF25-(GSH)2 because it is more because of the association with glutathione and has a very good water solubility,after the transformation EF25-(GSH)2 yellow powder from the EF25 changed into a white powder substance in Displaying dissolved in pure water is colorless solution,this feature makes the EF25-(GSH)2 has better prospects.In this study,cells were further discussed at the level of individual animals curcumin analogues EF25-(GSH)2 anti-hepatoma effect.First,we use different concentrations of EF25-(GSH)2 in vitro treatment of liver cancer cells and normal liver cells and H22 tumor-bearing mice,MTT cell viability assay,electron microscopy and laser scanning confocal microscopy to observe cell morphology,protein immunoblotting(Western blot)to detect AMPK/Akt/mTOR pathway-related protein expression changes.Then,to investigate the EF25-(GSH)2 tumor effect of the role of autophagy level and dose,but also the use of autophagy inhibitors late chloroquine(CQ)and EF25-(GSH)2 interaction in HepG2 cells and H22 tumor-bearing mice,testing relevant pharmacological targets.The results showed that EF25-(GSH)2 on HepG2 for 48h IC50 of 7.2 μmol/L,its growth inhibitory effects of curcumin and cisplatin was superior and less lethal effect on normal cells.Electron microscopy and laser scanning confocal microscope cell autophagy obvious phenomenon,a large number of autophagic vacuoles and cytoplasmic vacuoles phenomenon can clearly see from the pictures.HepG2 cells,the cell cycle is detected by flow cytometry after drug treatment,and found the occurrence of apoptotic cells.Tumor-bearing mice experiments showed that after administration found that mice EF25-(GSH)2 treatment groups in the administration of the Netherlands during the slow tumor volume growth from just before the administration 200mm3 to 370mm3,even effect superior to cisplatin(550mm3).At the same time the control group rats with curcumin group and subcutaneous tumor volume increases to close the rat model 1000mm3,and almost no toxicity in mice.Under CQ participation in moderate concentration(5-10μgmol/L)when the killing effect of the drug on the tumor cells more clearly,the individual animal experiments coincided with a small dose of the mutual authentication EF25-(GSH)2-CQ joint suitable concentration cancer treatment may have better results.Western blot results suggest EF25-(GSH)2 may be through AMPK/Akt/mTOR pathway-related inhibition of tumor cell growth.,revealing the EF25-(GSH)2 may be involved in tumor cell energy metabolism through relevant pathways causing tumor mechanism of cell killing effect,causing cells to produce a large number of autophagy,the formation of vacuoles structure within the cytoplasm huge loss of nutrients environmental pressures forced the cells died.Our results also proved EF25-(GSH)2 has good as liver cancer therapy development prospects.