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.