J Korean Cancer Assoc.  1997 Feb;29(1):11-18.

Expression and Intracellular Localization of Hepatitis C Viral Core Protein in Human Hepatoma Cell Line Transfected with Viral cDNA

• Kim DW
• Kim SH

Abstract

PURPOSE: Establishment of the human liver cell lines which permanently express the HCV proteins is important for the large scale production of viral antigen and analysis of the mechanism of hepatocellular carcinogenesis by HCV. Here, we attempted to establish the human hepatoblastoma cell lines which stably express the HCV core protein and examined the intracellular localization of the core protein.
MATERIALS AND METHODS
The cDNA of HCV core protein and neomycin resistance gene were expressed in HepG2 cells by the SRalpha promoter and human EF-1alpha gene promoter, respectively. The core protein was detected by immunofluorescence assay and western blotting.
RESULTS
We obtained several HepG2 cell clones which express HCV core protein stably. In transient expression assay, the core protein was localized in the cytoplasm in about 90%, and localized in the nucleus in about 10% of the core-expressing HepG2 cells. But, in the stably expressing HepG2 cell clones, the core protein was localized only in the cytoplasm. No HepG2 cell containing core protein in the nucleus was found in all of the cells which stably express the core protein.
CONCLUSION
The EF-1alpha gene promoter is highly efficient in the colony formation by neomycin resistance gene and is very useful for the isolation of human liver cell clones which express foreign genes stably. HCV core protein is localized in both nuclear and cytoplasm of human liver cell in short term but the cells containing the core protein in nucleus seem to disappear in long term culture.

Keyword

HCV core protein; Human hepatoblastoma cell line; Stable expression; Protein localization

MeSH Terms

Blotting, Western
Carcinogenesis
Carcinoma, Hepatocellular*
Cell Line*
Clone Cells
Cytoplasm
DNA, Complementary*
Fluorescent Antibody Technique
Hep G2 Cells
Hepatitis C*
Hepatitis*
Hepatoblastoma
Humans*
Liver
Neomycin
Peptide Elongation Factor 1
DNA, Complementary
Neomycin
Peptide Elongation Factor 1