摘要
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 explore the effect of Src tyro- sine kinase inhibition on phosphorylation of extracellular signal- regulated kinase 1 and 2 (ERK1/2) in non-small cell lung cancer (NSCLC) cells and its role. METHODS: NSCLC cells were cul- tured with different concentrations of Src tyrosine kinase inhibi- tor. Western blot was used to evaluate the phosphorylation of ERK1/2 in NSCLC cells and the effect of Src tyrosine kinase inhibition on phosphorylation of ERK1/2 in NSCLC cells. MTT was used to examine the effect of inhibition of Src tyrosine kinase on proliferation of NSCLC cells in vitro. Soft agar assay was used to assess the effect of Src tyrosine kinase inhibition on colony formation of NSCLC cells. RESULTS: ERK1/2 was phosphorylated in all NSCLC cell lines used in this study. Src tyrosine kinase inhibi- tion could suppress ERK1/2 phosphoryiation of PC-9 and A549 cells in a dose-dependent manner. Submicromolar Src tyrosine ki- nase inhibitor nearly suppressed phosphorylation of ERK1/2 in PC-9 and A549 cells entirely. Src tyrosine kinase inhibitor sup- pressed cell proliferation of PC-9 and A549 cells in a dose-depend- ent manner (F=5. 072, P=0. 004; F=4. 368, P=0. 008). The 0.1, 0.3 and 1.0 μmol/L Src tyrosine kinase inhibitor suppressed cell proliferation of PC-9 and A549 ceils by 14. 7~~, 47. 1~/00, 61.3G and 19. 8~/00, 24. 20/00, 30. 6~/00, respectively. While the same concentration of Src tyrosine kinase inhibitor had no significant effect on ERK1/2 phosphorylation in H226, PC14PE6 and RERFLCOK cells, as well as cell proliferation of H226, PC14PE6 and RERFLCOK cells. Src tyrosine kinase inhibitor significantly suppressed colony formation of PC-9 and A549 ceils (t= 11. 746, P〈0. 001; t= 5. 237, P〈0. 001). CONCLUSION: Src tyrosine kinase inhibition can suppress cell proliferation of PC-9 and A549 cells, which is attributed to the inhibition of ERK1/2 phosphorylation.
出处
《中华肿瘤防治杂志》
CAS
2011年第15期1150-1153,共4页
Chinese Journal of Cancer Prevention and Treatment
基金
辽宁省教育厅高等学校科研项目(20061011)
辽宁省自然科学基金(20082076)
辽宁省科技厅科学技术计划项目(2010225028)