Effect Of 5-fluorouracil On The Expression Of Smad7 And TβRI Of Human Keloid Fibroblasts In Vitro

Objective:It is to investigate the effect of 5-FU on the expression of Smad7 and TβRI mediated by TGF-β1 factor of human keloid fibroblasts in vitro.Material:The samples were prepared from six cases(equally in the amount of the two gender) of post-operation who had keloid respectively and the age of them ranges from 10-40 years old.Three of the prepared samples were originated from the up extremity,and the rest of the samples were stem from their faces.All of them didn't have the history of systemic disease and injection of hormone,and also,all of the patients agreed for the purpose of our experiment by using their body tissue.Methods:The experiment was conducted in Microbes Department of Anhui Medical University between Feb.2007 and Oct.2007.①Fibroblasts culture from primary cells:six cases of keloid were harvested to culture fibroblasts according to the document method,cell passaged after fusing.②The fourth to sixth generation cells were collected and seeded on the 96-hole plate,and cell viability after interfered by 5-FU for different concentration(10, 20,40,80,160)μM and different time(24,48,72)hour was evaluated by MTT assay to obtain the minimum concentration and the best time of cytotoxic drug.③According to the concentration and time of 5-FU which obtained in②,the effect of 5-FU on the expression of Smad7 and TβRI mediated by TGF-β1 factor in keloid fibroblasts were detected with Western blot.Results:①All the fibroblasts grew well and kept normal shape.Cell appearance kept the normal condition after interfered by 5-FU.②5-FU did not affect cell viability at 24h at the concentration of 10 and 20μM.(P＞0.05)③Compared with control group,the expression of Smad7 was obviously decrease and the expression of TβRI was obviously raise mediated bv TGF-β1 factor(P＜0.01);5-FU could significantly raised the expression of Smad7,and the highest expression was at the concentration of 20μM(P＜0.01);Meanwhile,5-FU did not show any effect on the synthesis of TβRI after TGF-β1 factor stimulation.Conclusion:5-FU can raise the expression of Smad7 mediated by TGF-β1 factor;but did not show any effect on the synthesis of TβRI after TGF-β1 factor stimulation.