| Budget Amount *help |
¥8,800,000 (Direct Cost: ¥8,800,000)
Fiscal Year 2005: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 2004: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 2003: ¥4,600,000 (Direct Cost: ¥4,600,000)
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| Research Abstract |
BACKGROUND & AIM : Hepatocyte growth factor (HGF) is a multifunctional growth factor, which is very effective for liver regeneration. On the other hand HGF has unfavorable "scattering" function which promotes cancer cell invasion and metastases. Recently HGF-MSP chimera, Metron Factor-1 (MF-1) was engineered to have favorable effects of HGF, being separated from the adverse effect of invasiveness. We examined biological effects of MF-1 in vitro and in vivo, and try to determine whether MF-1 can be a useful and safe regenerative factor for livers. METHODS : MF-1 protein was purified by two-step affinity chromatography. For in vitro study hepatocytes were exposed to MF-1 or HGF, and mitogenesis, Fas-mediated apoptosis, signal transduction and cell cycles was examined. Matrigel invasion assay was also performed using human HCC cell lines. For in vivo study, acute CCl_4 liver injury was induced in mice together with treatment of MF-1 or HGF, and the livers were histologically analyzed. In
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addition, HCC cells, transduced with HGF or MF-1 gene, were inoculated into livers, and intrahepatic metastasis was analyzed. RESULTS : When hepatocytes were exposed by MF-1 or HGF, phospholylated AKT and ERK1/2 were detected at 10 min, peaked at 30 min and decreased gradually afterward. ^3H-thymidine incorporation was increased with 2.4 folds by MF-1 and 3.3 folds by HGF comparing to that of control. Expression of cell cycle markers Cyclin D1 and Cdk4 were increased in the same manner by the treatments of both factor. Concerning anti-apoptotic function, MF-1 suppressed Fas-mediated apoptosis by 78%, while HGF suppressed by 41%. Moreover MF-1 did not affect HCC invasion through matrigel, while HGF promoted significantly. In vivo study, liver damage and apoptosis were significantly suppressed by MF-1 or HGF treatment group. At the same time intrahepatic metastasis were not induced by MF-1,though HGF promoted significant metastasis of HCC cells. CONCLUSION : MF-1 has favorable functions of proliferation and anti-apoptosis on hepatocytes, and no adverse effect of invasiveness and metastasis on HCC cells. These results suggested MF-1 can be a safe and useful growth factor for liver regeneration. Less
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