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目的探讨表皮生长因子(EGF)对真皮成纤维细胞中cyclinD1和CDK-4表达的影响,从细胞周期的角度认识EGF促进真皮成纤维细胞生长的机制。方法研究对象为大鼠真皮成纤维细胞,用含100ml/L胎牛血清的DMEM,加入50mg/L的EGF培养SD大鼠的真皮成纤维细胞,通过MTT检测、流式细胞仪分析观察细胞的生长状态,免疫组化检测cyclinD1和CDK-4的表达结合流式细胞仪分析来观察细胞的周期变化。结果MTT检测(加药前0.37±0.011,加药后0.55±0.008)、流式细胞仪结果(加药前9.1,加药后14.7)都显示50mg/L的EGF能显著促进真皮成纤维细胞的生长增殖,免疫组化结果显示,两者一致,加药后阳性明显增强。结论50mg/L的EGF对真皮成纤维细胞的生长有极大地促进作用,促进了细胞周期蛋白cyclinD1和CDK-4的表达增强,细胞G1期变短,增殖能力增强。
Objective To investigate the effect of epidermal growth factor (EGF) on the expression of cyclinD1 and CDK-4 in dermal fibroblasts, and to understand the mechanism of EGF in promoting the growth of dermal fibroblasts from the perspective of cell cycle. Methods The rat dermal fibroblasts were cultured in vitro. The dermal fibroblasts of SD rats were cultured in DMEM containing 100ml / L fetal bovine serum and 50mg / L EGF. MTT assay and flow cytometry Growth status, immunohistochemistry detection of cyclinD1 and CDK-4 expression combined with flow cytometry analysis to observe the cell cycle changes. Results MTT assay (0.37 ± 0.011 before dosing and 0.55 ± 0.008 d after dosing) showed that EGF (50 mg / L) could significantly promote the proliferation of dermal fibroblasts Growth and proliferation, immunohistochemistry results show that the two are consistent, significantly increased after dosing. Conclusion EGF (50mg / L) can greatly promote the growth of human dermal fibroblasts, promote the expression of cyclinD1 and CDK-4, shorten the G1 phase and enhance the proliferation of dermal fibroblasts.