O120 A functional assessment of hepatic ischaemic injury on hepatocellular carcinoma behaviour

Abstract

Abstract Introduction Hypothermic oxygenated perfusion (HOPE) preservation improves outcomes after donation after cardiac death (DCD) liver transplantation. Database analysis has also suggested significantly lower hepatocellular carcinoma (HCC) recurrence rates in recipients with HOPE-DCD livers compared to non-perfused livers. Whilst HOPE may protect against severe hepatic ischaemia-reperfusion injury, the exact mechanisms for lower recurrence rates are not fully understood. As HOPE preservation fluid, or perfusate, dynamically flushes out many inflammatory and ischaemic markers from the donor liver, the effect of these contents on in-vitro HCC cell behaviour was investigated. Methods An HCC cell line, HepG2, was functionally assessed following treatment with HOPE perfusate samples from 3 human livers declined for transplant. In-vitro HepG2 proliferation, stemness and initiation were assessed by an MTS assay, cytokeratin-19 (K-19) immunocytochemistry, and a colony-forming assay respectively. All experiments were conducted with approval by an ethical committee. Results All 3 HOPE perfusate samples resulted in significant HepG2 proliferation and increased aggressive behaviour potential. At 24 hours, HepG2 proliferation was significantly greater after treatment with 1:2 perfusate dilution compared to blank perfusate (p<0.05). Membrane invadopodia development, signifying an increased invasive potential, was also noted at 24 hours with treatments of 1:2 and 1:20, without observed differences in K-19 expression compared to untreated controls. Conclusion End-timepoint HOPE perfusate from declined livers facilitate increased HepG2 proliferation and aggression. HOPE may reduce early tumour recurrence in HCC patients by dynamically flushing out these liver-secreted trophic factors.