HCV-related hepatocellular carcinoma: gene signatures associated with TERT promoter mutations and sex

Abstract

Background: Hepatocellular carcinoma (HCC) is a rapidly progressing disease, frequently caused by hepatitis C virus (HCV) infection and a higher prevalence in males than females. Over 60% of HCCs harbour frequent activating mutations in the telomerase reverse transcriptase promoter (TERTp). However, the relationship between TERTp status, sex, and expression of specific genes remains poorly understood.

Methods: We conducted a literature search to identify genes that were significantly upregulated in HCV-related HCC, compared to the respective peri-tumour tissues, in at least two independent studies. We identified 90 genes and validated their expression in 59 matched HCV-related HCC and peri-tumour tissues using a custom multiplex array qPCR. HCV-related HCC patients were stratified by TERTp mutations and sex. Statistical analysis was performed to identify relationships between different variables.

Results: Overall, validation analysis confirmed the upregulation of 39 out of 90 genes. Expression levels of 24 genes differed significantly between HCV-related HCC with mutant and wild type TERTp. The expression of FASTK and FLVCR1 genes correlated with TNM and tumour size (p < 0.05). High expression of NUCKS1 was associated with mortality, particularly in male patients. Overall, the expression of 57 genes was sex-linked, with 26 and 48 genes significantly overexpressed in males and in females, respectively, some of which were also associated with mutant TERTp status.

Conclusions: We identified unique molecular signatures in TERTp mutant HCC associated with the activation of specific genes. Such results suggest that TERTp mutant HCC might represent a distinct clinical entity. Furthermore, the up-regulation of several genes in HCV-related HCC is sex-linked. These results are crucial for understanding the mechanisms underlying TERTp mutations in tumour progression and sex-related HCC risk and for developing more effective diagnostic and prognostic biomarkers.

Keywords: TERT; Biomarker; Female; Gene signatures; HCV; Male; Sex.

Fig. 1

PRISMA flow chart of the reports selected for the study. Ninety-five records were identified. 32 records were excluded because they did not meet the inclusion criteria. Therefore, 30 reports were considered relevant for full-text review and eligibility, of which 15 were included in this study

Fig. 2

DEGs in comparing HCV-related HCC versus peri-tumour tissues. This figure represents the mRNA expression levels of differentially expressed genes with ****P-value < 0.0001, ***P-value < 0.001, **P-value < 0.01, and *P-value < 0.05. P = peri-tumours; T = tumours

Fig. 3

DEGs in comparing TERTp mutant HCV-related HCC versus peri-tumour tissues. The mRNA expression levels of differentially expressed genes with ****P-value < 0.0001, ***P-value < 0.001, **P-value < 0.01, and *P-value < 0.05 are shown. P = peri-tumours; M = TERTp mutant tumours

Fig. 4

Venn diagram from a comparison of differentially expressed genes. DEGs in HCV-related HCC TERTp mutant versus peri-tumours were compared with DEGs in HCV-related HCC TERTp wild type versus peri-tumours. The numbers in circles represented the common DEGs expressed genes in both conditions (n = 6), not expressed in HCV-related HCC TERTp mutant (n = 4), and expressed in HCV-related HCC TERTp mutant samples (n = 24). P = peri-tumours; T = tumours; TERTpm = TERT promoter mutant tumours; TERTp WT = TERT promoter wild type tumours

Fig. 5

ROC curve and AUC. ROC curve of NUCKS1 (Std. error = 0.053; CI:95%; Lower limit = 0.756, Upper limit = 0.963, P-value < 0.0001), and FLVCR1 (Std. error = 0.061; CI:95%; Lower limit = 0.695, Upper limit = 0.934, P-value = 0.0003), in TERTp mutant HCV-related HCC. The corresponding AUC is reported in each area