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

这项研究的目的是评估辐射在有或没有EGFR突变的人类非小细胞肺癌（NSCLC）细胞中诱导的迁移（A549；野生型，HCC827；突变体类型）。进一步的目的是研究辐射诱导的细胞迁移在阻断EGFR及其下游途径后通过吉非替尼蛋白的迁移的作用。使用伤口愈合测定法评估了A549细胞和HCC827Cellswas的细胞迁移。通过WST-1分析和TUNEL分析测量细胞生长和凋亡。细胞周期扰动和EGFR信号转导通过流通仪和蛋白质印迹分析。 HCC827细胞的迁移距离显着降低了GEFITINIB和/或辐照，与未经处理的细胞相比，48小时或更长时间后，用吉非替尼和/或辐射升高了细胞HCC827细胞的死亡。 Thewst-1分析表明，A549细胞对A549细胞的行为没有差异。使用或不使用吉法替尼后2小时，使用特异性抗体辐照裂片的特异性抗体，通过蛋白质印迹揭示了凋亡途径的变化。通过在用gefitinib处理的HCC827细胞中裂解PARP的增加，凋亡途径的激活得到了证实。G2/M期停滞和HCC827细胞的吉非替尼和辐照治疗组中可见细胞周期中SUBG1的增加。吉非替尼和/或辐照组合处理可以通过下调HCC827细胞中ERK 1/2蛋白的表达来抑制ERK的磷酸化比率。在24h，48h或72h的时间过程中，TUNEL方法未检测到凋亡。因此，吉非替尼降低了EGFR基因突变的细胞中早期迁移粘附能力和早期凋亡。 Gefitinibshow在HCC827细胞中产生了细胞毒性作用并抑制细胞迁移。酪氨酸激酶受体抑制剂，用放射疗法进行的gefitinibcombcbbbin可能对NSCLC细胞具有细胞毒性，具有EGFR的遗传突变，随后抑制其细胞行为，包括增殖，入侵性，侵入性和转移活性。酪氨酸激酶受体抑制剂靶向的放射治疗方案可能为预防早期侵袭和晚期肺癌转移提供新的治疗方法。较少的