| Objective:To explore biological effects of 5-fluorouracil on the proliferation and apoptosis of human keloid fibroblasts in vitro.Methods:①Keloids of ear lobe,thigh,chest and upper arm were derived from 6 cases of Department of Plastic Surgery,First Affiliated Hospital of Anhui Medical University. Fibroblasts were derived from four keloids.TheⅣ-Ⅷgeneration exponential phase of growth cell were used for experiment.②Keloid fibroblasts were stimulated with 5-fluorouracil(0.1,0.2,0.4,0.8,and 1.6 g/L) for24,48,and 72 hours.The inhibition ratio and 50%inhibitory concentration(IC50) were calculated by MTT.In addition,0.2,0.3,0.4, 0.5 and 0.6 g/L 5-fluorouracil were prepared for the following experiment.③The cell apoptosis and cell cycle were measured by flow cytometry.④The cell morphological character was observed by Hoechst 33258 fluorescent staining.⑤The influence of 5-fluorouracil on Bcl-2 and Bax expressions were detected by western-blot.Results:Six keloid specimens were primarily cultured well,and all were included in the final analysis.①After 5-fluorouracil(0.1,0.2,0.4,0.8,and 1.6 g/L) intervention for 24,48, and 72 hours,the inhibition ratios were significantly different between every two groups(P＜0.01).The IC50 partly was(0.816±0.041),(0.637±0.028),(0.430±0.032) g/L by mapping.②After 5-fiuorouracil(0.2,0.3,0.4,0.5,and 0.6 g/L) intervention for 48 hours, the keloid fibroblasts apoptosis rate had significant differences between every two groups (P＜0.05).Compared with the control group ceils,the differences were statistically significant(P＜0.01).Compared with the control group cells,the number of experiment cells at phase G0/G1 significantly increased and S decreased(P＜0.05).③By Hoechst 33258 fluorescent staining,the keloid fibroblasts interfered with 5-fluorouracil showed nuclear condensation and apoptosis at different extents.④There was a remarkable decrease of the Bcl-2 expression and a marked increase of the Bax expression in different concentrations 5-fluorouracil compared with the control group(P＜0.05 ).Conclusion:The research revealed advanceedly the mechanism of action on 5-fluorouracil treated scars.5-fluorouracil could inhibit proliferation and induce apoptosis on human keloid flbroblasts in vitro.Low concentration and short-period injection with 5-fluorouracil would be a new consider of keloid treatment.
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