Resveratrol Induces Human Melanoma A375 Cells Apoptosis Through PI3K/AKT/mTOR Signaling Pathway

Background:Melanoma is a hot and difficult spot in the field of can-cer research,characterized by high invasiveness,high malignancy,and hi-gh drug resistance rate.Not only the pathogenesis is closely related to th-e PI3K/AKT/mTOR signaling pathway,but also its therapeutic and therp-eutic resistance is also closely related to this pathway.The main treatmen-t of early melanoma is still surgical method,the latter is mainly radiothe-rapy,chemotherapy,immunotherapy,ect.However,these treatments hav-e many side effects,such as treatment resistance,drug resistance,high co-st and other shortcomings,ect.As a result,many natural plant extracts w-hich have the advantages of cheap,effective and low side effects are con-cerned by the researchers.Among them,resveratrol as a natural antitoxin is widely studied at present because of a variety of anti-tumor effects.It h-as been proved that the molecular mechanism of resveratrol inducing the cancer based on PI3K/AKT/mTOR signaling pathway,such as gastric can-cer,liver cancer,chondrosarcoma and so on.There is no report about wh-ether the mechanism of resveratrol against melanoma is based on the PI3-K/AKT/mTOR signaling pathway.Object:To investigate the molecular mechanism of resveratrol induc-ing human melanoma A375 cells apoptosis based on PI3K/AKT/mTOR signaling pathway,to provide experimental and theoretical basis for the clinical application of resveratrol in the treatment of melanoma.Methods:The human melanoma A375 cells were treated with resver-atrol(0 umol/L?25 umol/L?50 umol/L?100 umol/L and 200 umol/L)aft-er 24 hours.The effects of resveratrol on human melanoma A375 cells pr-oliferation were detected by CCK-8 assay,and calculated IC50 value of 24 hours.The morphological changes of human melanoma A375 cells we-re detected by microscope.The apoptosis rate was detected by flow cyto-metry using Annexin V-FITC/PI double labeling method.The effects of resveratrol on apoptosis associated protein Bcl-2?Bax and PI3K/AKT/mT-OR signaling pathway associated protein PTEN?AKT?p-AKT?mTOR?p-mTOR expression were detected by Western Blot.Results:1.Resveratrol induced human melanoma A375 cells apop-tosis.The human melanoma A375 cells were treated with resveratrol(0 umol/L?25 umol/L?50 umol/L?100 umol/L and 200 umol/L)after 24 hours.Resveratrol inhibited human melanoma A3 75 cells proliferation significantly in a dose-dependent manner with CCK-8,IC50 value of 24 hours was 81.41?mol/L.Observed under optical microscope,the number of human melanoma A375 cells was reduced significantly,the gradual emergence of suspension cells and refraction was getting worse.After Annexin V/PI staining,with increasing concentrations of resveratrol,the apoptosis rate increases from(3.38±3.92)%to(62.94±4.67)%significa-ntly in a dose-dependent detected by flow cytometry.2.Resveratrol induced PI3K/AKT/mTOR signaling pathway inhibition on human mela-noma A3 75 cells.After treated with resveratrol(0 umol/L?25 umol/L?50 umol/L?100 umol/L and 200 umol/L)on human melanoma A375 cells for 24 h,Western Blot detection demonstrated that,with the increase of the concentration of resveratrol the protein level of pro apoptotic protein Bax expression was increased,the apoptosis inhibitory protein Bcl-2 exp-ression was decreased,the ratio of Bcl-2/Bax was decreased.With the in-crease of the concentration of resveratrol,PTEN protein expression was increased,p-AKT and p-mTOR protein expression was decreased,AKT and mTOR protein expression showed no significant change.Conclusions:1.Resveratrol significantly inhibits human melanoma A375 cell proliferation and induced apoptosis in a dose-dependent mann-er.2.Resveratrol may induce human melanoma A375 cells apoptosis by inhibiting the PI3K/AKT/mTOR signal transduction pathway.