Analysis of the Intracellular Zinc in HCV Replicon

Abstract

Aim: In patients with chronic liver injury, metabolic disturbance of zinc was frequently observed and Hepatitis C virus (HCV) replication is crucially involved with zinc metabolism. In vitro HCV replication system, HCV replicon enabled us to analyze HCV replication in vitro.
Here we aimed to quantitatively analyze the zinc in a HCV-infected hepatocyte, namely HCV replicon and control.
Methods: Genome-length HCV RNA-replicating cells, namely replicon cells (HCV-O cells) and control cells were treated with zinc salts and assayed their zinc content by in-air micro PIXE analyzer. Metallothionein (MT) which is a major reserve of zinc was also analyzed by
Cd-hem assay.
Results: 1) Micro-PIXE analysis revealed that zinc concentration was increased more in HCV-O cells than control. Additional zinc by zinc chloride administration enhanced the peak of zinc in both control and HCV replicon cells. Furthermore, the degree of zinc increment by
additional zinc is more in HCV-O cells than control. 2) Cd-hem assay also demonstrated the increase of zinc in HCV-O cells and zinc stimulation further increased the metallothionein content in HCV-O cells.
Conclusion: In-air-micro-PIXE and Cd-hem assay revealed the increase of zinc and metallothionein in HCV-O. Metallothionein increased and additional zinc also increased more in HCV-O. Thus, HCV infection increased zinc at least partially through metallothionein.