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目的探讨EGF信号通路影响雄激素非依赖型前列腺癌细胞系DU145细胞周期分子学机理。方法MTT法检测EGF对DU145细胞增殖能力的影响,流式细胞仪检测细胞周期,West-ern印迹检测细胞周期调控因子的表达情况。结果EGF组细胞增殖能力显著升高,EGF组细胞在S期细胞比例为65.36%,明显高于对照组的44.32%,两者比较差异有统计学意义(P<0.01);在蛋白质水平,EGF降低了p27表达,而CDK2、磷酸化Rb的表达不同程度增高,p16、p21表达无明显差异。结论EGF刺激导致其下游的信号通路活化,降低p27表达,并导致下游CDK2、Rb及Rb磷酸化水平的变化,最终促进了DU145细胞的增殖能力。
Objective To investigate the molecular mechanism of EGF signaling affecting the cell cycle of androgen-independent prostate cancer cell line DU145. Methods MTT assay was used to detect the effect of EGF on the proliferation of DU145 cells. The cell cycle was detected by flow cytometry and the expression of cell cycle regulators was detected by West-ern blot. Results The proliferation of EGF group was significantly increased. The percentage of cells in S phase in EGF group was 65.36%, which was significantly higher than that in control group (44.32%, P <0.01). At the protein level, EGF Decreased the expression of p27, while the expression of CDK2 and phosphorylated Rb increased to different extents, while the expression of p16 and p21 showed no significant difference. Conclusion EGF stimulation leads to the activation of its downstream signaling pathway, decreasing the expression of p27 and the phosphorylation of CDK2, Rb and Rb in the downstream, and eventually promoting the proliferation of DU145 cells.