Miyata Hideki, Sasaki Tamito, Kuwahara Kenichi, Serikawa Masahiro, Chayama Kazuaki
Department of Medicine and Molecular Science, Division of Frontier Medical Science, Programs for Biochemical Research, Graduate School of Biomedical Sciences, Hiroshima University, 1-2-3 Kasumi, Hiroshima 734-8551, Japan.
Int J Oncol. 2006 Apr;28(4):915-21.
The signaling pathway that is initiated by binding of epidermal growth factor receptor (EGFR) and results in sustained signaling through PI3K plays an important role in a tumor's response to ionizing radiation. The current in vitro study explored both the effects of ZD1839 (Iressa), a highly selective EGFR tyrosine kinase inhibitor, as a radiosensitiser for bile duct carcinoma cell lines and ZD1839's general effects on cell growth in the same two lines. Secondly, we ensured suppression of radiation-induced phosphorylation of EGFR by ZD1839 using an immunoprecipitation technique. Furthermore, we examined radiation-induced phosphorylation of ERK, p38, JNK, and AKT with or without inhibitor with use of Western blot techniques and performed clonogenic assays to confirm radiosensitivity in the presence of a drug. ZD1839 inhibited cell growth of both cell lines and suppressed radiation-induced phosphorylation of EGFR. After exposure to radiation, there was an increase in phosphorylation of AKT as shown by Western blot. Treatment with either ZD1839 or LY294002 (the latter, a PI3K inhibitor) suppressed phosphorylation of AKT by Western blot. Both ZD1839 and LY294002 significantly suppressed colony formation by clonogenic assay; however, U0126 (a MEK1/2 inhibitor), SB203580 (a p38 inhibitor), and SP600125 (a JNK inhibitor) had no effect on colony formation. These results suggest that AKT may be a useful target molecule for enhancement of radiotherapy effect and that ZD1839 may have an important role in combination with radiotherapy for patients with bile duct carcinoma.
由表皮生长因子受体(EGFR)结合所启动并通过PI3K导致持续信号传导的信号通路,在肿瘤对电离辐射的反应中起重要作用。当前的体外研究探讨了ZD1839(易瑞沙),一种高度选择性的EGFR酪氨酸激酶抑制剂,作为胆管癌细胞系放射增敏剂的效果以及ZD1839对同一两种细胞系中细胞生长的总体影响。其次,我们使用免疫沉淀技术确保ZD1839抑制辐射诱导的EGFR磷酸化。此外,我们使用蛋白质印迹技术检测有无抑制剂时辐射诱导的ERK、p38、JNK和AKT的磷酸化,并进行克隆形成试验以确认药物存在时的放射敏感性。ZD1839抑制两种细胞系的细胞生长并抑制辐射诱导的EGFR磷酸化。蛋白质印迹显示,暴露于辐射后,AKT的磷酸化增加。用ZD1839或LY294002(后者为PI3K抑制剂)处理可通过蛋白质印迹抑制AKT的磷酸化。ZD1839和LY294002均通过克隆形成试验显著抑制集落形成;然而,U0126(MEK1/2抑制剂)、SB203580(p38抑制剂)和SP600125(JNK抑制剂)对集落形成无影响。这些结果表明,AKT可能是增强放疗效果的有用靶分子,并且ZD1839在胆管癌患者的放疗联合治疗中可能具有重要作用。