BACKGROUND: Hepatocellular carcinoma (HCC) is a fatal disease characterized by early genetic alterations in telomerase reverse transcriptase promoter (TERTp) and β-catenin (CTNNB1) genes and immune cell activation in the tumor microenvironment. As a novel approach, we wanted to assess patient survival influenced by combined presence of mutations and densities of CD8+ cytotoxic T cells. METHODS: Tissue samples were obtained from 67 HCC patients who had undergone resection. We analysed CD8+ T cells density, TERTp mutations, rs2853669 polymorphism, and CTNNB1 mutations. These variables were evaluated for time to recurrence (TTR) and disease free survival (DFS). RESULTS: TERTp mutations were found in 75.8% and CTNNB1 mutations in 35.6% of the patients. TERTp mutations were not associated with survival but polymorphism rs2853669 in TERTp was associated with improved TTR and DFS. CTNNB1 mutations were associated with improving TTR. High density of CD8+ T-lymphocytes in tumor center and invasive margin correlated with longer TTR and DFS. Combined genetic and immune factors further improved survival showing higher predictive values. E.g., combining CTNNB1 mutations and high density of CD8+ T-lymphocytes in tumor center yielded HRs of 0.12 (0.03-0.52), p = 0.005 for TTR and 0.25 (0.09-0.74), p = 0.01 for DFS. CONCLUSION: The results outline a novel integrative approach for prognostication through combining independent predictive factors from genetic and immune cell profiles. However, larger studies are needed to explore multiple cell types in the tumor microenvironment.

Bioptická laboratoř s r o Mikulášské nám 4 326 00 Pilsen Czech Republic

Department of Cancer Epidemiology German Cancer Research Center Im Neuenheimer Feld 280 69120 Heidelberg Germany

Department of Histology and Embryology Faculty of Medicine in Pilsen Charles University Karlovarska 48 301 66 Pilsen Czech Republic

Department of Pathology 3rd Faculty of Medicine Charles University Ruská 87 100 00 Prague 10 Czech Republic

Department of Surgery Faculty of Medicine in Pilsen Charles University Alej 16 Svobody 80 323 00 Pilsen Czech Republic

Laboratory of Cancer Treatment and Tissue Regeneration Biomedical Center Faculty of Medicine in Pilsen Charles University Prague Czech Republic

Laboratory of Pharmacogenomics Biomedical Center Faculty of Medicine in Pilsen Charles University Pilsen Czech Republic

Laboratory of Quantitative Histology Biomedical Center Faculty of Medicine in Pilsen Charles University Alej Svobody 1665 76 323 00 Pilsen Czech Republic

Laboratory of Translational Cancer Genomics Biomedical Center Faculty of Medicine in Pilsen Charles University Alej Svobody 1665 76 323 00 Pilsen Czech Republic

Sikl's Institute of Pathology Faculty of Medicine and Teaching Hospital in Plzen Charles University Plzen Czech Republic

Toxicogenomics Unit National Institute of Public Health Prague Prague Czech Republic

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miRNA and lncRNA as potential tissue biomarkers in hepatocellular carcinoma