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目的:探讨抑制Src酪氨酸激酶活化对非小细胞肺癌(NSCLC)细胞外信号调节激酶的影响及其作用。方法:采用NSCLC细胞株进行细胞培养,分别给予不同浓度的Src酪氨酸激酶抑制剂。蛋白质印迹检测NSCLC细胞外信号调节激酶1/2(ERK1/2)磷酸化以及抑制Src酪氨酸激酶活化对NSCLC细胞ERK1/2磷酸化的影响。MTT法检测抑制Src酪氨酸激酶活化对NSCLC细胞体外增殖的影响。软琼脂糖集落形成实验检测抑制Src酪氨酸激酶对NSCLC细胞克隆形成的影响。结果:选用的NSCLC细胞都存在ERK1/2磷酸化。抑制Src酪氨酸激酶对PC-9和A549细胞ERK1/2磷酸化呈现浓度依赖性抑制作用,亚微摩尔水平Src酪氨酸激酶抑制剂几乎完全抑制PC-9和A549细胞ERK1/2磷酸化。Src酪氨酸激酶抑制剂对PC-9和A549细胞体外增殖表现出明显的浓度依赖性抑制作用(F=5.072,P=0.004;F=4.368,P=0.008)。0.1、0.3和1.0μmol/L的Src酪氨酸激酶抑制剂对PC-9和A549细胞增殖的抑制率分别为14.7%、47.1%、61.3%和19.8%、24.2%、30.6%。而其余3种NSCLC细胞ERK1/2磷酸化以及体外增殖对Src酪氨酸激酶抑制剂反应不敏感。Src酪氨酸激酶抑制剂明显抑制PC-9和A549细胞的克隆形成(t=11.746,P<0.001;t=5.237,P<0.001)。结论:抑制Src酪氨酸激酶能够抑制PC-9和A549细胞ERK1/2磷酸化,从而抑制PC-9和A549细胞体外增殖。
Objective: To investigate the effect of Src tyrosine kinase inhibition on extracellular signal-regulated kinase in non-small cell lung cancer (NSCLC). METHODS: NSCLC cell lines were used for cell culture and Src tyrosine kinase inhibitors were given at different concentrations. The effects of ERK1 / 2 phosphorylation and Src tyrosine kinase activation on the phosphorylation of ERK1 / 2 in NSCLC cells were detected by Western blotting. Inhibition of Src tyrosine kinase activation on the proliferation of NSCLC cells in vitro by MTT assay. Effect of inhibiting Src tyrosine kinase on colony formation of NSCLC cells by soft agarose colony formation assay. RESULTS: ERK1 / 2 phosphorylation was present in all selected NSCLC cells. Inhibition of Src tyrosine kinase showed a concentration-dependent inhibition of ERK1 / 2 phosphorylation in PC-9 and A549 cells, and sub-micromolar Src tyrosine kinase inhibitors almost completely inhibited ERK1 / 2 phosphorylation in PC-9 and A549 cells . Src tyrosine kinase inhibitors showed a significant concentration-dependent inhibition of proliferation in vitro in PC-9 and A549 cells (F = 5.072, P = 0.004; F = 4.368, P = 0.008). The inhibitory rates of Src tyrosine kinase inhibitor 0.1, 0.3 and 1.0 μmol / L on the proliferation of PC-9 and A549 cells were 14.7%, 47.1%, 61.3% and 19.8%, 24.2% and 30.6%, respectively. However, ERK1 / 2 phosphorylation and proliferation in the other three NSCLC cells were not sensitive to Src tyrosine kinase inhibitors. Src tyrosine kinase inhibitors significantly inhibited the clonogenicity of PC-9 and A549 cells (t = 11.746, P <0.001; t = 5.237, P <0.001). Conclusion: Inhibition of Src tyrosine kinase can inhibit the phosphorylation of ERK1 / 2 in PC-9 and A549 cells and inhibit the proliferation of PC-9 and A549 cells in vitro.