Direct Reprogramming HCC Cell To Liver-like Cell By Small Molecule Compounds

Research background Hepatocellular carcinoma(HCC) is the second leading cause of cancer-related mortality worldwide. In China, where nearly half of world's newly diagnosed cases and death cases occurred, liver cancer is the third most common cancer. Till today,surgical resection remains the primary and most effective treatment method for early-stage HCC patient. However, vast majority of patients have already lost opportunity for surgery at time of diagnosis. Other treatments, including melting,TACE, chemotherapy as well as newly emerged immunotherapy and target therapy(precision medicine), have not yet achieve significant efficacy in HCC treatment due to the disease's heterogeneity and high tendency of drug-resistance. Therefore, finding new treatment approach, new idea and new technology for HCC controlling or abrogation remains a critical mission.Induce cell reprogramming is a new technology, it refers to a cell type switching. In the reprogramming process, cell biological behaviors, gene expression profile, and epigenetic state are dramatically changed. The successful HCC cell reversion to liver-like cell using reprogramming approach may provide a new possibility to control even cure liver cancer.Research methods1. To investigate the effect of SMC on cell apoptosis induction of HCC cell SMMC-7721 and Hep G2 by flow cytometry.2. To examine the effect of SMC on tumor behavior suppression by CCK8, cell cycle,Edu, invasion, migration, plate cloning formation, soft-agar cloning formation and animal assay.3. To test whether reprogrammed HCC cell SMMC-7721 and Hep G2 obtained normal liver cell feature, functions and expression of liver cell related genes by immunostaining assay, Elisa assay, PAS, Oil red staining and RT-PCR assay.4. To study the underlining molecular mechanism of SMC reprogramming HCC cell SMMC-7721 and Hep G2 to liver-like cell by Western blot.Result1. The reprogramming process was accompanied with HCC cell apoptosis.2. The tumor behaviors of reprogrammed HCC cell SMMC-7721 and HepG2 have been suppressed.2.1 The morphological change.2.2 The arrested cell growth ability.2.3 The inhibition of cell metastasis and invasion ability.2.4 The reprogrammed HCC cell lost colony formation potential when growing in vitro.2.5 The reprogrammed HCC cell lost tumorigenicity potential in vivo.3. The reprogrammed HCC cell obtained normal liver cell feature, functions and expressed liver cell related genes.4. Tumor related signal pathways were strongly inhibited by SMC treatment.ConclusionUsing small molecular compounds to regulate cell signaling and epigenetic state,HCC cells could be reverted to normal liver-like cells with the loss of tumorigenicity potential in vitro and in vivo and this process was accompanied with cell apoptosis induction. Our research suggests that the cell reprogramming technology could be applied to the liver cancer treatment research. Our research results would provide a new idea and approach for HCC treatment study and may provide a new possibility for effective controlling even curing HCC.