Expression And Clinical Significance Of CISD2 In Hepatocellular Carcinoma

Objective: The present study aimed to explore the role and clinical significance of CISD2 in hepatocellular carcinoma(HCC)and the potential mechanism of CISD2 expression in HCC.Methods:(1)Immunohistochemical staining was used to detect the protein expression level of CISD2 in 100 HCC tumor tissue samples and 46 para-carcinoma normal liver tissue samples,and to analyze the correlation between CISD2 expression and clinicopathological features,proliferation-related protein ki-67,and 5-year overall survival rate of the patients.(2)Western blot was used to detect the expression of CISD2 in human hepatoma cell lines BEL7404,Hep G2,SMMC-7721 and human normal liver cell line L02,and to analyze the expression differences.The stable low CISD2 expression HCC cell lines BEL7404 and SMMC-7721 were constructed,and the effects of CISD2 on the proliferation,invasion,and metastasis of HCC cells were detected by CCK8 and cell migration assay.(3)Through bioinformatics analysis,online web tool STRING was used to construct the CISD2 protein interaction network,and gene ontology enrichment was used to analyze CISD2 protein function;Web TIMER was used to analyze the relationship between CISD2 and the infiltration of immune cells in hepatocellular carcinoma.Kaplan-Meier-plotter analyzed the relationship between CISD2 and 5-year survival rates in patients with different groups of hepatocytes.GESA analyzed the signaling pathways involved in the high expression of CISD2 in hepatocellular carcinoma.Results:(1)The expression level of CISD2 in HCC tumor tissues was significantly higher than that in para-carcinoma liver tissues,and the expression level of CISD2 in HCC cell lines was also significantly higher than that in normal liver cells,with statistically significant differences(P<0.01).(2)CISD2 expression was positively correlated with tumor size in HCC patients(P<0.05).(3)CISD2 expression was significantly positively correlated with the ki-67 percentage of patients' clinicopathological data(P<0.01).(4)The overall survival rate of patients with high CISD2 expression was lower than those with low CISD2 expression.(5)Bioinformatics analysis showed that in patients with no history of alcohol consumption or hepatitis virus infection,the overall survival rate of those with high CISD2 m RNA expression was lower than those with low CISD2 m RNA expression.(6)CISD2 was negatively correlated with the infiltration of B cells,CD4+ T cells,CD8+ T cells,macrophages,and dendritic cells.(7)CISD2 is mainly involved in intracellular Fe2+ iron and sulfur group binding,iron and sulfur cluster binding and metal cluster binding.(8)GESA analysis showed that CISD2 high expression promoted porphyrin and chlorophyll metabolism,pentose and glucuronic acid conversion,ascorbic acid and aldehyde acid metabolism,amino and nucleotide glucose metabolism,and starch and sucrose metabolism in hepatocellular carcinoma.Conclusion:(1)CISD2 is highly expressed in hepatocellular carcinoma,and its expression level is related to the prognosis of patients.(2)High expression of CISD2 promotes the proliferation and metastasis of hepatocellular carcinoma,and CISD2 may become a prognostic marker and a potential therapeutic target for hepatocellular carcinoma.(3)CISD2 may promote the survival and proliferation of tumor cells by affecting their iron ions,energy metabolism and tumor immune microenvironment.