Effect of gefitinib on radiation-inducedmigration in human lung cancer cells.

本研究的目的是评价辐射是否诱导有和无EGFR突变（A549 ;野生型，HCC 827;突变型）的人非小细胞肺癌（NSCLC）细胞迁移。进一步的目的是研究吉非替尼体外阻断EGFR及其下游通路后对辐射诱导的细胞迁移的影响。采用创伤愈合实验检测A549细胞和HCC 827细胞的迁移能力。分别用WST-1法和TUNEL法检测细胞生长和凋亡。流式细胞术和Western blotting分析细胞周期紊乱和EGFR信号转导。吉非替尼和/或照射可显著缩短HCC 827细胞的迁移距离，并使照射48小时或更长时间后HCC 827细胞的死亡率增加。WST-1法显示，A549细胞在不同浓度下的行为无明显差异， ...更多信息 HCC 827细胞对增加吉非替尼浓度敏感72小时或更长时间。使用特异性抗体和裂解的PARP，在加或不加吉非替尼的情况下照射2小时后，通过Western印迹法揭示了凋亡途径的变化。吉非替尼处理后，肝癌细胞PARP裂解增加，细胞周期阻滞于G2/M期，细胞周期亚群G1期增加。吉非替尼和（或）照射联合作用可通过下调ERK 1/2蛋白的表达，抑制ERK磷酸化水平。TUNEL法检测24 h、48 h、72 h时程均未检测到细胞凋亡。因此，吉非替尼降低了EGFR基因突变细胞的早期迁移粘附能力和早期凋亡。吉非替尼对HCC 827细胞具有细胞毒作用，并抑制细胞迁移。酪氨酸激酶受体抑制剂吉非替尼联合放疗可能对EGFR基因突变的NSCLC细胞具有细胞毒性，随后抑制其细胞行为，包括增殖、侵袭和转移活性。以酪氨酸激酶受体为靶点的联合放疗方案可能为预防晚期肺癌的早期侵袭和转移提供新的治疗方法。少

The aim of this study was to evaluate whether radiation induces migration in human non-small cell lung cancer(NSCLC) cells with and without an EGFR mutation(A549 ; wild type, HCC827 ; mutant type). A further aim was to investigate the effects ofradiation induced cell migration by the cells after blocking the EGFR and its downstream pathways by gefitinibin vitro. Cell migrationof A549 cells and HCC827cellswas assessed using a wound healing assay. Cell growth and apoptosis were measured by the WST-1 assay and TUNEL assay, respectively. Cell cycle perturbation and EGFR signal transduction were analyzed by flow-cytometry and Western blotting. The migration distance of the HCC827 cells was significantly decreased bygefitinib and/or irradiation, and the death of the cells HCC827 cells wasincreased with gefitinib and/or irradiation after 48hours or more in comparison to untreated the cells. There was no difference inthe behavior of A549 cells with the treatment.TheWST-1 assay showed that the … More HCC827 cells were sensitive to increasing concentrationsof gefitinib for 72 hours or more. A variation in the apoptotic pathway was revealed by Western blotting using specific antibodies with cleaved PARP irradiated 2hours after with or without gefitinib. The activation of the apoptosis pathway was confirmed by increased cleaved PARP in HCC827 cells treated with gefitinib.G2/M phase arrest and increased subG1 in cell cycle was seen in the combination gefitinib and irradiation treatment group of HCC827 cells. The gefitinib and/or irradiation combination treatment could inhibit the phosphorylated ratio of ERK by down-regulating the expression of ERK 1/2 proteins in HCC827 cells. No apoptosis was detected by the TUNEL method in the time course of 24h, 48h, or 72h. Consequently, gefitinib reduced the early migration adhesion ability, and early apoptosis in cells with a genetic mutation of EGFR. Gefitinibshowed a cytotoxic effect and inhibited cell migration in HCC827 cells. The tyrosine kinase receptor inhibitor, gefitinibcombined with radiotherapy may be cytotoxic to NSCLC cells with a genetic mutation of EGFR with subsequent inhibition of their cellular behavior, including proliferation, invasiveness, and metastatic activity. The tyrosine kinase receptor inhibitor-targeted combined radiotherapy regimen may provide a new treatment forthe prevention of early invasion and metastasis for advanced lung cancer. Less