Risk of Development, Outcome, and New Tumor Markers of a Hepatocellular Carcinoma

Abstract

Background: HCV infection and its complications, especially hepatocellular carcinoma, is a substantial public health burden. In 2015 “Nationwide hepatitis C elimination program” was launched in Georgia. According to the protocol, patients with HCC also receive DAA antiviral treatment. Many studies shows that Gankyrin is one of the main players in hepatocellular carcinogenesis. It expression increased not only in HCC tissues, but also in cirrhotic livers, while Gankyrin absent in normal liver samples

Aim: The aim of this study was to define the effect of the different DAA therapy regiments on the incidence or recurrence of HCC and its prognosis and to investigate the feasibility of plasma circulating Gankyrin mRNA as a HCC biomarker.

Methods and Materials: Overall, 408 patients were recruited between April 2015-March 2016. The selection criteria were as follows: age 50-65 years; Liver fibrosis level F3-F4 or cirrhosis at least 15 years of disease history; HCV positive diagnosed by PCR method; 4. absence of previous complications of cirrhosis. Child-Pugh class A or B; and absence of severe extrahepatic disease.  Clinical monitoring and management of adverse events were performed on a regular base. HCV All patients included in the study received anti-HCV treatment with direct-acting antivirals (DAAs) within the national hepatitis C elimination program in accordance with national protocols. During April 2015-March 2016 treatment was provided with sofosbuvir (SOF) in combination with ribavirin (RBV), with or without pegylated interferon (IFN). Since March 2016, ledipasvir/sofosbuvir (LDV/SOF) was prescribed to all patients with or without RBV depending on the HCV genotype, level of fibrosis, and previous treatment experience. Incidence and mortality rates per 100 person-years (PY) of follow-up were calculated as number of events divided by the total person-years of observation multiplied by 100. Bivariate comparisons were tested by Pearson’s chis-square or Fisher exact test as appropriate. Factors associated with the occurrence of HCC were evaluated in Cox proportional hazards regression model. All analyses were conducted using SAS v9.4. P values of <0.05.

Plasma samples were isolated from 4 mL of peripheral blood from patient with HCC (n=32), HCC with MTS  (n=5), Cirrhosis - (n=7), HCV positive - (n=5), and healthy individuals. Fresh Total DNA/RNA/miRNA was extracted from blood plasma using RecoverAll™ Total Nucleic Acid Isolation Kit. RT-qPCR was performed using TaqPath 1-Step Multiplex Master Mix. Human 18S ribosomal RNA was used as an internal reference gene. The relative expression levels were calculated using 2−ΔΔCq method using SAS-Studio Mann-Whitney U test was used to determine the statistical significance for comparisons between groups.

Results: we find that neither different DAA regimens nor treatment duration affects HCC risk after antiviral treatment. There are no significant changes in mortality rate due to HCC in these groups. Gankyrin RNA expression is significantly increase in  plasma of HCC patients, especially in metastatic diseases. 

Conclusion: Therefore, it can be concluded, that HCC status is not a contraindication for DAA treatment, especially at the early stages of cancer, when a tumor is curative. Plasma circulating Gankyrin mRNA is high sensitive HCC biomarker for liquid biopsy.