Development of gene therapy for metastatic liver cancer from colorectal cancer by introduction of Axin gene and TCF decoy using HVJ-E

点击翻译按钮获取中文摘要

1.AIM :The aim of this study was to investigate the alteration in the Wnt signaling pathway in hepatocellular carcinoma and colorectal carcinoma and to develop of the treatment of colorectal cancer by regulation of Wnt signaling pathway using Axin gene.2.RESULTS(1)Immunohistochemisty of β-catenin and mutational analysis of β-catenin and Axin1 in colorectal cancer : High frequency of accumulation of β-catenin was observed in colorectal cancer. Furthermore, there were several mutations in Axin1 gene. (2)Construction of metastatic liver cancer model in rat : Because the metastatic model previously developed was not applicable, we used subcutaneous tumor model using nude mouse. (3)Construction of TCF/β-catenin dependent Axin1 expression vector (TOP-Axin). (4)Introduction of TOP-Axin to culture cells : TOP-Axin and PEF-BOS/myc-Axin were transfected to COS-7 cells by lipofectamine. Myc-Axin expression was clearly detected by western blotting using anti-myc and anti-Axin antibodies. However, TOP-Axin expression was not enough detected. Although GSK3-β inhibitor induced accumulation of β-catenin, TOP-Axin expression was not induced. We decided to examine the effect of PEF-BOS/myc-Axin on colon cancer in animal model. (5)Gene transfection using HVJ-E : GFP expression vector was introduced to the culture cells, introduction efficiency was approximately 10%-20%, and it couldn't obtained enough efficiency. (6)Introduction of Axin gene to animal model by HVJ-E : The expression of Axin couldn't be confirmed by immunohistochenistry using anti-Axin and anti-myc antibodies when PEF-BOS/myc-Axin vector was injected to subcutaneous tumor using HVJ-E.3.Future view and now on going :We are going to construct Axin-Sendai virus vector to use in animal model in vivo.