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

吉非替尼对辐射诱导的人肺癌细胞迁移的影响。

• 批准号: 22791168
• 负责人: SATO Yumi
• 金额: $ 1.08万
• 依托单位: 埼玉県立がんセンター (2011)Gunma University (2010)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2010
• 资助国家: 日本
• 起止时间: 2010 至 2011
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-22791168/
• 关键词: 遊走能; ゲフィチニブ; 肺癌; 放射線

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

本研究的目的是评估辐射是否会诱导有或没有 EGFR 突变（A549；野生型，HCC827；突变型）的人类非小细胞肺癌 (NSCLC) 细胞的迁移，进一步的目的是研究其影响。体外用吉非替尼阻断 EGFR 及其下游通路后，辐射诱导细胞迁移。使用伤口愈合试验评估细胞生长和凋亡。分别通过WST-1测定和TUNEL测定测定细胞周期扰动，并通过流式细胞术和Western印迹分析HCC827细胞的迁移距离显着缩短，并且细胞死亡。与未处理的细胞相比，使用吉非替尼和/或照射48小时或更长时间后，HCC827细胞有所增加。治疗后 A549 细胞的行为存在差异。WST-1 测定表明，HCC827 细胞对增加浓度的吉非替尼敏感达 72 小时或更长时间，通过使用具有裂解的特异性抗体的蛋白质印迹法揭示了细胞凋亡途径的变化。在使用或不使用吉非替尼的情况下照射 PARP 2 小时通过处理的 HCC827 细胞中裂解的 PARP 的增加证实了细胞凋亡途径的激活。吉非替尼联合放疗组的HCC827细胞中观察到G2/M期阻滞和细胞周期subG1增加。吉非替尼和/或放疗联合治疗可以通过下调ERK的表达来抑制ERK的磷酸化比例。 TUNEL法在24h时程内未检测到HCC827细胞中ERK 1/2蛋白的凋亡。 48h或72h检测，吉非替尼降低了EGFR基因突变细胞的早期迁移粘附能力，吉非替尼在HCC827细胞中表现出细胞毒作用并抑制细胞迁移，吉非替尼与放疗联合治疗可能是有效的。对具有 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、侵袭性和酪氨酸激酶受体抑制剂靶向联合放疗方案可能为预防晚期肺癌早期侵袭和转移提供新的治疗方法。