ERK阻滞剂U0126对白血病细胞K562细胞周期的作用及机制

Effect of ERK pathway inhibitor U0126 on cell cycle of leukemia K562 cell lines

目的研究细胞外信号调节激酶(extracellular signal-regulated kinase,ERK)阻滞剂U0126对K562细胞周期的作用及机制。方法以逆转录-聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)和Westernblot检测U0126处理K562细胞后ERK、Cyclin D2、Cyclin E、P27 mRNA和蛋白的表达;以流式细胞术(flow cytometry,FCM)检测细胞周期的变化。结果处理后K562细胞ERK、Cyclin D2、Cyclin E mRNA和蛋白表达降低;P27 mRNA和蛋白表达增高。G0/G1期细胞增多,S期细胞减少,与用药前相比差异均有统计学意义(P<0.05)。结论U0126可能通过ERK途径,影响细胞周期调控蛋白,最终抑制了K562细胞的增殖。

Objective To study the mechanism of extracellular signal-regulated kinase （ERK） pathway inhibitor U0126 on cell cycle of K562 cell lines. Methods The mRNA and protein expressions of ERK, Cyclin D2, Cyclin E in and P27 K562 cell lines after 10 μmol/L U0126 treatment for 12, 24 and 72 h were detected by RT-PCR and Western blot, respectively. The cell cycle was determined by flow cytometry. Results The mRNA and protein expressions of ERK, Cyclin D2 and Cyclin E in K562 cell lines after U0126 treatment were decreased, while those of P27 were increased. The percentage of cells in G0/G1 phase was increased and that in S phase was decreased. All data mentioned above showed significant difference between the K562 cell lines before and after U0126 treatment （ P 〈 0. 05 ）. Conclusion U0126 possibly affects cell cycle regulating proteins by ERK pathway and eventually inhibits the proliferation of K562 cells.