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

这项研究的目的是评估辐射是否诱导带有和不带有EGFR突变(A549；野生型，HCC827；突变型)的人非小细胞肺癌(NSCLC)细胞的迁移。进一步研究吉非替宾体外阻断EGFR及其下游通路后，细胞辐射诱导的细胞迁移效应。用创伤愈合实验检测A549细胞和HCC827细胞的细胞迁移。WST-1法检测细胞生长，TUNEL法检测细胞凋亡率。用流式细胞仪和Western blotting分析细胞周期变化和EGFR信号转导。Gefitinib和/或照射显著缩短了HCC827细胞的迁移距离，48h或更长时间照射后细胞死亡明显增加。WST-1检测显示，…对A549细胞有明显的抑制作用更多的HCC827细胞对增加浓度的吉非替尼72小时或更长时间敏感。用切割后的PARP与Gefitinib或不加Gefitinib照射2小时后，用免疫印迹法检测到细胞凋亡途径的变化。Gefitinib诱导的HCC827细胞G2期/M期停滞，细胞周期中亚G1期增多。吉非替尼和/或放射联合作用可通过下调细胞内ERK 1/2蛋白的表达来抑制ERK的磷酸化比例。TUNEL法在24、48、72小时均未检测到细胞凋亡。因此，吉非替尼降低了带有EGFR基因突变的细胞的早期迁移黏附能力和早期凋亡。吉非替尼对HCC827细胞有明显的细胞毒作用和细胞迁移抑制作用。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细胞产生细胞毒作用，从而抑制其细胞行为，包括增殖、侵袭和转移活性。酪氨酸激酶受体抑制剂-靶向联合放射治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案。较少

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