| ObjectiveTo construct hepatic progenitor cells (HPCs) expressing hepatitis B virus X (HBx) and establish a mouse model by portal vein injection of the cells,in order to provide a platform to study the effect of HBx on HPCs and the mechanism of hepatocellular carcinoma (HCC).MethodsThe HPCs (cell line 14-19) were transfected by recombinant adenovirus vectors expressing HBx gene (Ad-HBx-GFP) as well as empty vector (Ad-GFP) separately,and the transfected HPCs were infused to the mouse from the portal vein. The control group was given equivalent normal saline. The change of serum alanine aminotransferase (ALT) was detected on the 3rd、7th and 14th days. The expression of HBx and green fluorescent protein (GFP) in liver tissue was detected by immunohistochemical technique and frozen section method. The expression of HBx at mRNA and protein levels was measured by RT-PCR and Western blotting. The expression of cyclin D1 at mRNA and protein levels on 7th days was measured by RT-PCR and Western blotting.ResultsThe mouse model expressing HBx was successfully established.1 The GFP expression,serum ALT level and HBx-positive cells reached the peaks on the 7th day. Compared with the control group and the 14-19/Ad-GFP group, thel4-19/Ad-HBx-GFP group showed specific expression of HBx mRNA and protein in the liver tissue.2 Compared with the control group and the 14-19/Ad-GFP group, the14-19/Ad-HBx-GFP group showed high expression of cyclin D1 mRNA and protein in the liver tissue on the 7th day.Conclusion1 The 14-19 cells expressing HBx and protein GFP is successfully.2 The mouse model expressing HBx is successfully established with reliable operation and good repeatability,3 The 14-19/Ad-HBx-GFP group showed high expression of cyclin D1 mRNA and protein in the liver tissue on the 7th day. It may prove that HBx can promote the proliferation of hepatic stem cells.That lays a basis for exploring the mechanism of HCC and provides a new thought for establishing a tumor animal model. |
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