摘要
目的：观察酪氨酸激酶抑制ARQ197对肺癌H1299细胞的放射增敏作用及机制。方法首先用不同浓度的重组人肝细胞生长因子（ HGF）和ARQ197分别处理H1299细胞，应用克隆形成实验法检测细胞增殖，筛选出用于放射敏感性研究的HGF和ARQ197的合适浓度。随后将细胞分为对照组、HGF处理组、ARQ197处理组、HGF＋ARQ197处理组，观察不同组别之间差异。最后用蛋白印记检测HGF、单纯放射或联合应用ARQ197对c-Met及下游Akt和Erk 1／2蛋白表达和活化的影响。结果H1299细胞的克隆形成率随着HGF浓度增加呈进行性升高，而ARQ197则进行性下降。 LQ模型细胞存活曲线示HGF、HGF＋ARQ197处理及ARQ197对H1299细胞的放射增益比分别为0-85、1-20、1-27（存活分数为0-1时的剂量比）。 H1299细胞在 HGF 刺激后 p-cMet、p-Akt、p-Erk 1／2高表达， HGF＋ARQ197中随着ARQ197浓度增加p-cMet、p-Akt、p-Erk 1／2蛋白表达进行性下降。细胞接受2 Gy照射后p-cMet、p-Akt、p-Erk 1／2高表达，但照射＋ARQ197后p-cMet、p-Akt、p-Erk 1／2蛋白表达显著下降，但总的cMet、Akt、Erk 1／2蛋白表达无明显变化。结论 ARQ197通过抑制HGF／c-Met及其下游传导通路的活化对离体肺癌H1299细胞有显著放射增敏作用。
Objective To evaluate the radiosensitizing effect and action mechanism of a tyrosine kinase inhibitor, ARQ197, on lung cancer cell line H1299. Methods H1299 cells were treated with different concentrations of recombinant human hepatocyte growth factor （ HGF） and ARQ197, respectively. Colony formation assay was used to investigate the proliferation of H1299 cells, and the optimal concentrations of HGF and ARQ197 were figured out for the radiosensitivity study. H1299 cells were divided into control, HGF-treated, ARQ197-treated, and HGF＋ARQ197-treated groups, and the differences between those groups were evaluated. The impacts of HGF, radiation alone, or HGF＋ARQ197 on the expression and activation of c-Met and the downstream Akt and Erk 1/2 were determined by Western blot. Results The colony formation rate of H1299 cells increased with increasing HGF concentration, and decreased with increasing ARQ197 concentration. Using the linear-quadratic model, the cell survival curve showed that the radiation dose enhancement ratios for H1299 cells treated with HGF, HGF＋ARQ197, and ARQ197 were 0-85, 1-20, and 1-27, respectively, when the surviving fraction was 0-1. The expression of pc-Met, p-Akt, and p-Erk 1/2 proteins increased in H1299 cells after HGF stimulation. In HGF＋ARQ197-treated cells, the expression of pc-Met, p-Akt, and p-Erk 1/2 proteins decreased with increasing ARQ197 concentration. After 2 Gy irradiation, the expression of pc-Met, p-Akt, and p-Erk 1/2 proteins increased in HGF-treated cells but substantially decreased in HGF＋ARQ197-treated cells, while there were no significant changes in the expression of total c-Met, Akt, and Erk 1/2 proteins. Conclusions ARQ197 has a substantial radiosensitizing effect on lung cancer cell line H1299 in vitro by inhibiting HGF/c-Met and activation of downstream signaling pathways.
出处
《中华放射肿瘤学杂志》
CSCD
北大核心
2015年第5期588-592,共5页
Chinese Journal of Radiation Oncology
基金
江苏省“科教兴卫”医学重点人才项目（RC2011144）
江苏省自然科学基金资助（BK20141185）
