STUDIES OF HEPG2 CELLS INFECTED WITH HGV RNA GENOME / 微生物学通报

ABSTRACT

In order to observe the replication and expression of HGV RNA genome in HepG2 cells and establish a cell model of HGV infection, HGV RNA genome was prepared in vitro and transfected HepG2 cells with lipofec-tamin. HGV RNA-positive supernatants were used to infect fresh HepG2 cells. RT-PCR, immunohistochemistry and Western blot assays were carried out to detect the replication and expression of HGV in HepG2 cells. Both positive and negative strands of HGV RNA could be detectable in cell culture supernatants and cells at 24h post-transfection. During the culture periods of 90 days, the cells were maintained by changing the medium every 3 or 5 days, and cultured for more than 20 passages. Both strands of HGV could be detectable in culture supernatants and cells. Immunohistochemistry and Western blot results also confirmed that HGV E2 protein could be expressed in the infected HepG2 cells. HGV RNA could also be detectable in the frozen-thawed HepG2 cells infected with HGV RNA genome. Therefore, HGV RNA genome can replicate and express in HepG2 cells, this HGV RNA genome transfected cells model could be used as a cell model in the studies of replication and infection of HGV.