Effect of VEGF and endostatin gene transfer on the time course expression of endoglin after partial hepatectomy in a mouse model

Abstract

hepatc, cyte-denved cells. We also examined the effect of bile acid on LST-2 gene transcription. Methods: 1) Expression of [be LST-2 gene was analyzed by northern blot analysis 2) The promoter actb~ty of the 5'-upstream region of LST-2 gene was analyzed by' luciferase reporter gene assays. 3) The specihc pmtein-DNA complex was determined by electoruphoresis mobility-shih assays (EMSA). 4) The effects of bile acids on the LS%2 promoter activity were analyzed by lnciferase assays and EMSA Results: Norther blot analysis showed that LST-2 was expressed in the hepatocyte-derived cell lines, Hep3B and HT-17. Deletion analysis showed that the 5'-tlanking region from -180 to -20 bp is responsible top LST-2 trans~npticmal actwity There were several putative transcriptional factor recognition sites for STAT, HNFIa, FXR, and two binding sites tbr HNF3b in this region. By site-directed mutation analysis, It was revealed that all of the consensus binding sites for FXR, HNFla, HNF3b play important roles m the transcriptional activity of the LST-2 gene. By EMSA using specilic oligonucleotides and mmlear extract of Hep3B, we observed specific pmteinDNA complex of HNF1, FXR, and HNF3b. Moreover, we observed the super-shift bands using specific antibodies against HNF1, FXR and HNF3h Next, we carried out the luciterase assays in the presence of bile acids. The /uciterase activity was increased to 2.0 [bids when CDCA or DCA was added, g~scussion: These results indicate that the transcriptional regulation of IST-2 is controlled by FXR, HNF1, and HNF3b. HNF1 and HNF3b might depend on the tissue-specific expression and FXR might play a role in transcriptional activation by CDC)~ and DCA Interestingly, the transcription of LST-2 gene is regulated by the substrate which itself is taken in.