Antitumor Effects And Molecular Mechanism Of TZY2,a Novel Naphthalimide Derivative,on Hepatocellular Carcinoma In Vitro And In Vivo

Objective:In this study,the antitumor effects and molecular mechanism of TZY2,a novel naphthalimide derivant,was examined in both Hep G2 cells and SMMC-7721 cells in vitro and tumor-bearing mice in vivo.Methods:The antiperliferative activity of TZY2 was examined by MTT assay.The morphological changes of cells were observed by HCS with AO/EB staining.Apoptotic rate were measured by Flow Cytometry with Annexin V/PI double staining.Cell cycle distribution after treatment with TZY2 was detected by Flow Cytometry with PI staining.We also detect the change of mitochondrial membrane potential by HCS and Flow Cytometry with Rh123 staining,respectively.DCFH-DA fluorescence probe was applied to detect the cellular reactive oxygen species?ROS?level.Wound scratch experiment was used to observe cell migration.The protein expression of PARP-1,Caspase-8,Bcl-2,p53 and other relative proteins by Western blot.Moreover,we explored antitumor effects of TZY2 using H22 solid tumor model,H22 lung metastasis model in KunMing mice and CT-26 lung metastasis model in BALB/c mice in vivo.Results:TZY2 significantly inhibited SMMC-7721 cells and Hep G2 cells growth in a dose-dependent manner,after 48 h treatment,the IC₅₀ was?3.3±1.01??M,?8.75±0.74??M,respectively.TZY2 could induce Hep G2 cells and SMMC-7721 cells apoptosis and decrease of mitochondrial membrane potential in a time-and concentration-dependent manner.Furthermore,TZY2 activated p53,PARP-1,Caspase-8 and Caspase-3,up-regulated the protein expression of Bax and down-regulated the protein expression of Bcl-2,and also promoted cytochrome C release from mitochondria to cytoplasm.Scratch experiments showed that TZY2inhibited cellular migration and down-regulated the expression of?-catenin.TZY2 also showed potent inhibitory effects on tumor growth and metastssis in H22 solid tumor growth in KunMing mice,H22 lung metastasis and CT-26 lung metastasis in BALB/c mice in vivo,the inhibition rate was 48.99%,61.80%and 63.92%,respectively.Conclusions:TZY2 exerted potent antitumor growth and metastasis activity in hepatocellular carcinoma in vitro and in vivo and these effects was related to TZY2-mediated cell apoptosis.