Screening,identification and preliminary functional analysis of TFAM differential binding proteins between hepatocellular carcinoma and adjacent non-cancerous tissues

ZHAO Xin; WU Dan; XING Jinliang; HE Xianli

doi:10.13276/j.issn.2097-1656.2025.07.016

Abstract：Objective To explore the differential characteristics of mitochondrial transcription factor A(TFAM)binding proteins during the occurrence and progression of hepatocellular carcinoma(HCC),and to offer a novel perspective and theoretical foundation for the pathogenic mechanism and therapeutic strategies of HCC.Methods The 4D-FastDIA quantitative proteomics technology was used to conduct a comprehensive analysis of HCC tissues and their paired adjacent non-cancerous tissue samples,screening out proteins that exhibit significant differences in binding with TFAM in HCC tissues.By means of bioinformatics,GO functional enrichment and KEGG pathway enrichment analyses were conducted for these differential binding proteins.At the cellular level,the interaction relationship between TFAM and key binding proteins was verified using co-immunoprecipitation and Western blotting.Moreover,the role of these key binding proteins in the proliferation of HCC cells was explored through EdU experiments.Results Proteomics analysis indicated that the proteins binding to TFAM in HCC cells were significantly enriched in signaling pathways related to cellular protein translation and translocation,purine and amide metabolic synthesis,cellular immune response and cell differentiation.Further,co-immunoprecipitation and Western blotting analyses demonstrated that in SNU-739 cells,TFAM interacted with anti-silencing function 1B histone chaperone(ASF1B),aldehyde dehydrogenase 1 family member L1(ALDH1L1),Ral-interacting protein(RBP1),BRG1-associated factor 180(PBRM1),and BRG1-associated factor 170(SMARCC2).In SNU-739 cells,interfering with the expression of ASF1B,RBP1,and SMARCC2 could inhibit cell proliferation(P＜0.01),while interfering with the expression of ALDH1L1 and PBRM1 promoted cell proliferation(P＜0.05).Conclusion Through preliminary screening and identification,it is discovered that TFAM-specific binding proteins ASF1B,ALDH1L1,RBP1,PBRM1,and SMARCC2 in HCC cells may become novel biomarkers in the occurrence and progression of HCC,and provide new research directions for the development of targeted therapeutic targets,which is of great clinical significance for improving the diagnosis,treatment and prognosis of HCC patients.