TRANSLATIONAL AND CLINICAL MEDICINE - Georgian Medical Journal

Introduction: Early diagnosis of hepatocellular carcinoma (HCC) is crucial for improving patient survival rates. Gankyrin/PSMD10 is significantly involved in the carcinogenesis of hepatocellular tumors. This study aims to explore the potential of plasma circulating gankyrin mRNA expression as a biomarker for HCC.

Methods: Blood samples were collected from patients with HCC (n = 32), metastatic HCC (n = 5), cirrhosis (n = 7), hepatitis C virus positive (HCV+; n = 5), and healthy controls (n = 5). Gankyrin/PSMD10 RNA expression was assessed in tumor tissue samples from HCC patients (tumor and adjacent tissues; n = 32), cirrhotic patients (n = 5), and normal tissues (n = 5). Plasma samples were isolated from 4 ml of peripheral blood. Total DNA/RNA/miRNA was extracted from the plasma using the RecoverAll Total Nucleic Acid Isolation kit. Sections of formalin-fixed, paraffin-embedded (FFPE) tissue, each up to 20 μm thick, were combined into a 1.5 ml microcentrifuge tube. Total DNA/RNA was extracted from the FFPE tissues of tumor, adjacent liver, and control samples using the same kit. RT-qPCR was conducted with TaqPath 1-Step Multiplex Master Mix, amplifying cDNA using the following TaqMan assays: p28/Gankyrin (Assay ID Hs01100439_g1) and human 18S ribosomal RNA (Assay ID Hs99999901_s1).

Results: Gankyrin/PSMD10 RNA was not detected in serum samples from patients with cirrhosis, HCV+, or healthy individuals. The mean ΔCq in the HCC group was 11.69 ± 1.6, while in patients with metastatic HCC, it was 10.16 ± 0.4. A statistically significant difference in ΔCq was observed between groups (P < 0.0002). Notable differences in gankyrin expression levels were found in tissue samples. The 2-ΔΔCq mean for gankyrin/PSMD10 in HCC, adjacent tumor tissue (HCC ADJ T), cirrhosis, and control groups were 418.5 ± 220.6, 78.68 ± 49.40, 20.62 ± 11.32, and 1.02 ± 0.23, respectively, with statistically significant differences between groups (P < 0.001).

Conclusions: This study provides compelling evidence that gankyrin RNA expression is significantly elevated in HCC compared to normal and cirrhotic tissues. Additionally, gankyrin RNA levels in tumor-adjacent liver tissue were higher than those in cirrhotic liver tissues of patients without HCC. Gankyrin RNA was undetectable in the blood plasma of healthy individuals and patients without cancer; however, its expression was significantly increased in patients with HCC. Detecting gankyrin RNA in liquid biopsies may be effectively utilized for screening at-risk patients for HCC development. While gankyrin is not specific to HCC, it should be used alongside other molecular markers for HCC diagnosis but can serve as a standalone marker for monitoring and early detection of tumor recurrence.