HBV X Gene Transfection Upregulates IL-1β and IL-6 Gene Expression and Induces Rat Glomerular Mesangial Cell Proliferation

摘要

The X gene of HBV encodes a 17-kD protein,termed HBx,which has been shown to function as a transcriptional trans-activator of a variety of viral and cellular promoter/enhancer elements.The aim of this study was to investigate the effect of HBx on gene expression of interleukin(IL)-1β and IL-6,and proliferation of rat mesangial cells in vitro.The X gene of HBV was amplified by PCR assay,and inserted into the eukaryotic expression vector pCI-neo.The structure of recombinant pCI-neo-X plasmid was proved by restrict endonuclease digestion and sequencing analysis.pCI-neo-X was transfected into cultured rat mesangial cell line in vitro via liposome.HBx expression in transfected mesangial cells was detected by Western blot.The IL-1β and IL-6 mRNA expression in those cells was assayed by semiquantitative RT-PCR.Mesangial cell proliferation was tested by MTT.The results showed that HBx was obviously expressed in cultured mesangial cell line at 36th and 48th h after transfection.The expression of IL-1β and IL-6 mRNA was simultaneously increased.The cell proliferation was also obvious at the same time.It was concluded that HBx gene transfection could induce IL-1β and IL-6 gene expression and mesangial cell proliferation.HBx may play a critical role in mesangial cell proliferation through upregulation of the IL-1β and IL-6 gene expression.