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目的探讨表皮生长因子(ep iderm al growth factor,EGF)对一膜表面稳定表达表皮生长因子受体(EGF receptor,EGFR)细胞系CHO-EGFR-GFP1生物学特性的影响。方法采用台盼蓝染色计数活细胞数观察不同浓度EGF刺激48 h后细胞增殖情况;PI染色一步法流式细胞仪检测不同浓度EGF刺激48 h和100 ng/mL EGF刺激不同时间细胞周期改变以及细胞凋亡,Ho-echst 33258染色观察细胞凋亡;流式细胞术测定GFP(green fluorescent prote in,绿色荧光蛋白)平均荧光强度判定EGFR表达水平的变化。结果低浓度的EGF可促进CHO-EGFR-GFP1细胞增殖,而高浓度EGF刺激则引起CHO-EGFR-GFP1细胞失去黏附、细胞周期阻滞、促进细胞凋亡以及抑制细胞增殖;高浓度EGF以一种剂量依赖性方式同时上调EGFR的表达和引起细胞凋亡。结论不同浓度EGF对CHO-EGFR-GFP1细胞生长特性影响不同,高浓度EGF刺激引起肿瘤细胞失去黏附可能是导致肿瘤细胞容易脱落、发生转移的机制之一。
Objective To investigate the effects of epidermal growth factor (EGF) on the biological characteristics of a stable expression of EGF receptor (EGFR) cell line CHO-EGFR-GFP1 on a membrane surface. Methods The number of viable cells was counted by trypan blue staining and the proliferation of cells was observed after 48 h of EGF stimulation. PI staining was used to detect the changes of cell cycle at different concentrations of EGF for 48 h and 100 ng / mL EGF, The apoptosis was detected by Ho-echst 33258 staining. The average fluorescence intensity of GFP (green fluorescent protein) was measured by flow cytometry to determine the change of EGFR expression. Results Low concentration of EGF could promote the proliferation of CHO-EGFR-GFP1 cells, while EGF-GFP1 cells lost the adhesion, cell cycle arrest, cell apoptosis and inhibited the proliferation of CHO-EGFR-GFP1 cells. In a dose-dependent manner, EGFR expression and apoptosis were both up-regulated. Conclusion Different concentrations of EGF have different effects on the growth characteristics of CHO-EGFR-GFP1 cells. The high concentration of EGF may cause the loss of adhesion of tumor cells, which may lead to the tumor cells shedding and metastasis easily.