[Primary study of HCV replication in HepG2 cells inoculated by anti-HCV positive and HCVRNA RT-PCR positive serum].
OBJECTIVE
To study HCV replication in HepG2 cell line and to establish a stable HepG2 cell line infected with HCV.


METHODS
Continuous inoculation of anti-HVC positive and HCVRNA RT-PCR positive sera was used to infect HepG2 cells. The plus and minus strains of HCVRNA were detected by reverse transcription polymerase chain reaction in the HepG2 cells. Virus particles were observed in postinoculated HepG2 cells by transmission electron microscope. Immunohistochemical staining was used respectively to detect HCV antigen, HCVNS5 and HVC capsid in postinoculated HepG2 cells.


RESULTS
HCV RNAs (both plus and minus strains) were found from the first passage to the third passage of postinoculation HepG2 cells, and HCV viruslike particles were detected in the third passage of the infected HepG2 cells, those particles mostly occurred within cytoplasm, their diameters were between 30-60 nm, and gathering silver particles were found in immunogold silver staining of HCV NS5/capsid protein in infected HepG2 cells. Gathering gold particles were found mostly within vesicles in cytoplasm by immuno gold electron microscopy.


CONCLUSIONS
HepG2 cell line infected by HCV is established by continuous inoculation of anti-HCV positive and HCVRNA positive sera. HCV can replicate and produce virus-like particles in HepG2 cells, which may occur in the serial passages of post-infected HepG2 cells.
Lai
L Y
LY
Department of Infectious Disease, Second Affiliated Hospital, Hunan Medical University, Changsha 410011.
Gong
G Z
GZ
Zeng
Q S
QS
D000914:Antibodies, Viral / Q000097:blood*; D016174:Hepacivirus / Q000502:physiology*; D018197:Hepatoblastoma / Q000821:virology; D006801:Humans; D007106:Immune Sera; D008113:Liver Neoplasms / Q000821:virology*; D012367:RNA, Viral / Q000097:blood; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured; D014779:Virus Replication
D000914:Antibodies, Viral; D007106:Immune Sera; D012367:RNA, Viral