The Influence Of Hepatitis B Virus X Protein And Its Truncated Mutants To Mouse Foetal Hepatic Progenitor Cells Proliferation

Background and objective:The hepatitis B virus x (HBx) protein is an important factor in hepatitis B virus (HBV)-associated hepatocellular carcinoma (HCC). The C-terminal region of HBx plays a major role in the replication of HBV. Notably, HBx promotes the expansion and tumorigenesis of hepatic progenitor cells (HPCs) in mice. However, it remains unclear whether the C-terminal region of HBx is required to stimulation proliferation and malignant potential in mouse foetal hepatic progenitor cells (FHPCs),this study will be focused on the effects of the C-terminal region of HBx on FHPCs proliferation.Methods:Joint differential centrifugation and density gradient centrifugation screening of hepatic progenitor cells. Identification the expression of CD 133, EPCAM, CD49f and CK19 by cell immunofluorescence in hepatic progenitor cells. Adding DMSO, HGF, according to periodic acid snow’s surname dyeing (PAS), cell immunofluorescence and flow cytometry, detection the hepatic progenitor cells differentiation into hepatic cells and bile duct epithelial cells. After transfection HBx protein, detection the cell transfection efficiency by PCR, observing the diversity of morphological, Proliferation and multiplication capacity in hepatic progenitor cells.Results:(1) We used EPCAM+, CD 133+ and CD49f+ FHPCs, which are bipotential clonogenic cells.These FHPCs transform into mature hepatocytes and cholangiocyte when cultured under conditions that facilitate differentiation.(2) Compared with FHPCs grown as monolayers, spherical cell proliferation occurs more quickly.(3) Spherically cultured FHPCs can grow in semi-solid agar and tend to maintain the morphology and characteristics of stem cells compared with growth in rat tail collagen.(4) The C-terminus of HBx stimulates the proliferation of FHPCs but is not necessary to form spheroids similarly to Hepatic Cancer Stem CellsConclusion:The results furthers understand of the HBx-induced tumourigenicity of FHPCs and will aid in the treatment of HCC.