FTH Renders The HCC Cells Specifically Resist To Ferroptosis By Maintaining Iron Homeostasis

Background: Hepatocellular carcinoma（HCC）belongs to the widespread primary liver malignancy,which is different from the secondary due to the leading cause of HCC has not yet been completely clarified at present.The difficulty in the hepatocytes of HCC has increased in a high degree.Currently,it is a promisig therapeutic strategy that induction of ferroptosis in Fe-rich cancer cells.Ferroptosis,a newly discovered type of iron-reliant,programmed cell death,which target preferentially to iron-rich cancer cells,such as HCC、NSCLC 、 PDAC and breast cancer.Ferritin heavy chain（FTH）,a key components of the iron storage nano-cage,which not only of ferroxidase activity but slao against the generation of iron-mediated ROS.Our previous studies have shown that FTH could increase cancer cells resistance to ferroptosis.Nevertheless,its precise mechanismic roles of ferroptosis-resistance maintains obscure in the occurrence and development of HCC.Objective: The main specific mechanism of this topic is exploring why FTH facilitates the proliferation and drives the HCC cells particularly against to ferroptosis.We aim to further the understanding of the therapeutic mechanisms that inducing ferroptosis HCC cells death,which supplies a reliable basis for further investidations of the molecular targeting therapy option for HCC.Methods: Bioinformatic analysis was conducted using the indicated online websites and databases.Cell viability of HCC was assessed by CCK-8;Cell proliferation、 migration and wound healing assays were used to analyze the capability of cell proliferation and migration.Next,the detection of ROS、LPO、SOD、MMP、mitochondrial morphology and free iron were performed using fluorescence probes including DCF-DA、C11-BODIPY（581/591）、mito SOX、mito Tracker 、 JC-10 、 TMRM and RPA.The OCR of mitochondria was monitored by Seahorse XFe 24 Bioanalyzer in real time.Protein expression was determined by western blot.The male BALB/c mice were used for vitro test to observe the body weight of the mice and the tumor volume,and then the tumor tissues were sectioned for H＆E staining and immunohistochemistry.Results: Bioinformatic analysis revealed FTH expression levels were higher in multiple carcinoma tissues（LIHC 、 CHOL 、 HNSC etc.）than in normal adjacent tissues.In addition,the serum ferritin concentrations were retrospectively analyzed between January 2012 and November 2020 in Zhejiang Provincal People’s Hospital,which were discovered to be positively associated with the progression of disease as those of chronic hepatitis、liver cirrhosis and HCC.The further study also indicated the expression of FTH was closely related to the grade 、 stage and prognosis of HCC.Importantly,protein interaction network has elucidated FTH is essential for maintenance of iron homeostasis and relies on lysosome-dependent degradation.On the other hand,the results of the basis cell experimental study also indicated that the high expression of FTH enhance both the HCC proliferation and migration,and play a spefic action on favouring HCC resist ferroptotic death.However,it did not protect the cell death elicited by oxaliplatin 、 irinotecan and adriamycin.Mechanistically,the upregulation of FTH in HCC is beneficial for reducing the accumulation of the hydroxyl peroxide,decreasing the levels of mitochondrial ROS,attenuating the mitochondrial respiratory chain dysfunction,and rescuing mitochondrial dynamic equilibrium.Remarkably,from the results of IHC,the overexpression of FTH increased the fluorescence intensity of PCNA,reduced the production of lipidperoxides,as well argmented HCC tumorigenicity in vivo.Conclusion: FTH as an oncogene plays an significant role in the occurrence and progression of HCC and is predicted to be a novel target for ferroptosis therapy.