Hepatic Carcinoma Cells Supernatant Promotes Fibroblasts To Express COX-2and HGF

Objective：to observe the proliferation and transdifferentiation offibroblasts and the expression of COX-2and HGF influenced by micehepatic carcinoma cells supernatant. To investigate whether fibroblastscould be used to solve the problem--low long-term survival rate ofhepatocytes after cell transplantation.Methods：the growth tendency of fibroblasts cultured by normalmedium, medium contained TGF-β1, and mice hepatic carcinoma cellssupernatant involved was detected by MTT assay. The mRNA and proteinexpression level of α-SMA、COX-2and HGF expressed by fibroblasts indifferent culture conditions were detected by RT-PCR andimmunohistochemistry respectively.Results：MTT assay indicated that conditional medium containedtumor cells supernatant and the medium contained TGF-β1couldobviously promote the proliferation of fibroblasts. But the conditionalmedium had stronger effect. The results of PCR showed conditionalmedium and medium contained TGF-β1had stronger effect on thetransition from fibroblasts to myofibroblasts and could maintain thephenotypic of myofibroblasts than normal medium did, and spurredmyofibroblasts to express COX-2and HGF. It was suggested that the expression of α-SMA reached the peak on day3and kept steadily. Theexpression of COX-2and HGF reached their peaks on day5and thenmaintain the level.α-SMA expressd by fibroblasts cultured by normalmedium reached its expression level peak on day5and decreased on day7.But Fibroblasts cultured by normal medium expressed no COX-2andHGF.Conclusions:it is suggested that fibroblasts could transdifferentiateto be myofibroblasts Which could express COX-2and HGF stably whencultured in the medium with mice hepatic carcinoma cells supernatant.Only through transdifferentiation could fibroblasts express COX-2andHGF. Further more, fibroblasts multiplicated notably and expressed moreCOX-2and HGF influenced by mice hepatic carcinoma cells supernatant.The results above laid the theoretical foundation for combinedhepatocytes and fibroblasts transplantation improving long-term survivalrate of hepatocytes.