The Mechanism Of MTFR2 In Affecting Prognosis And Immune Infiltration Of Hepatocellular Carcinoma

Objective: Hepatocellular carcinoma has the seventh highest incidence worldwide and the second highest mortality rate among cancers.Asia is the region with the second highest incidence of primary hepatocellular carcinoma.The prognosis of hepatocellular carcinoma is pretty poor because of the limitations of treatments.Fortunately,immune checkpoint inhibitors(ICI)have shown inspiring efficacy in hepatocellular carcinoma.However,only a few patients can benefit from an ICI treatment.Mitochondrial fission regulator 2(MTFR2)is involved in the progression and development of various cancers.However,the roles of MTFR2 in HCC remain largely unknown.In this study,through a comprehensive analysis of MTFR2 in HCC,we explored the relationship between MTFR2 and the prognosis and immune infiltration of HCC.Further analysis of the possible molecular mechanism,so as to find biomarkers to predict the immunotherapy response of HCC patients and potential targets to enhance the efficacy of immunotherapy,providing a new theoretical basis for the diagnosis and treatment of HCC.Methods:(1)Real-time fluorescence quantitative PCR(RT-QPCR)was used to detect the m RNA expression level of MTFR2 in 7 human HCC cell lines and human liver cell lines LO2.Western Blot was used to detect MTFR2 protein expression level in 8 pairs of HCC and adjacent tissues.(2)MTFR2-related gene data in HCC were obtained from GEO and TCGA databases,and the relationship between copy number variation(CNV)status and m RNA expression level was analyzed,as well as differentially expressed genes in groups with high MTFR2 expression level and low MTFR2 expression level.Meanwhile,KEGG and GO analysis was performed to explore its potential functions.(3)HCC surgical specimens were collected and immunohistochemical staining of MTFR2 was performed with tissue microarray to verify MTFR2 expression and its correlation with prognosis of HCC patients.(4)The correlation between MTFR2 expression and immune cell infiltration and immune-related molecule expression was analyzed using TIMER,GEPIA and TISIDB databases.(5)RNA-Seq data of anti-PD-1 immunotherapy cohort were used to analyze the effect of MTFR2 expression level on efficacy,and to explore the correlation between MTFR2 and immune checkpoint and immunotherapy prediction pathway.(6)The serum index data of anti-PD-1 immunotherapy patients were collected to analyze the correlation between MTFR2 and serum immune markers.Results:(1)The expression level of MTFR2 is up-regulated in HCC,leading to shorter OS and worse DFS in patients.(2)MTFR2 is related to a variety of immune functions and immune-related signaling pathways.(3)MTFR2 is positively correlated with the level of immune cell infiltration in the HCC tumor microenvironment.(4)MTFR2 is positively correlated with multiple immune checkpoint inhibitors and immunotherapy response prediction pathways.(5)The immunotherapy validation cohort analysis showed that in the patients with higher MTFR2 expression levels,the immune barrier can be broken and the patients can get better immunotherapy response.Conclusions:(1)MTFR2 expression level is an independent prognostic factor of HCC patients.(2)MTFR2 inhibits the anti-tumor T cell immune response of HCC,which is an influencing factor of poor prognosis of HCC patients.(3)MTFR2 is a potential biomarker of immunotherapy response,which can be used as a biomarker to predict the prognosis of HCC,providing a new theoretical basis for the diagnosis and treatment of HCC.