The synergistic effect of dimethylamino benzoylphenylurea (NSC #639829) and X-irradiation on human lung carcinoma cell lines
The synergistic effect of dimethylamino benzoylphenylurea (NSC #639829) and X-irradiation on human lung carcinoma cell lines
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DOI：
10.1007/s00280-006-0333-3
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发表时间：
2007-06-01
影响因子：
3
通讯作者：
Edelman, Martin J.
中科院分区：
文献类型：
--
作者：
Balcer-Kubiczek, Elizabeth K.;Attarpour, Mona;Edelman, Martin J.
目的 本研究旨在研究 N-[4-(5-溴-2-嘧啶氧基)-3-甲基苯基]-(二甲氨基)-苯甲酰基苯基脲 (二甲氨基苯甲酰基苯基脲；BPU) 使细胞对辐射敏感的能力，并检查非小细胞肺癌 (NSCLC) 细胞系中表型与存活、DNA 损伤、凋亡或细胞周期进展之间的关系。 方法 三种 NSCLC 的异步培养使用（表型）系、A549（腺癌）、NCI-H226（鳞状细胞）和NCI-H596（腺鳞癌）。用不同浓度（0-10μM）的BPU处理细胞24小时，以获得抑制细胞存活约50%（IC50）的药物剂量。细胞在没有 BPU 的情况下或在 BPU 处理 24 小时后以 IC50 进行 X 射线照射。辐射剂量范围为 0 至 10 Gy。通过集落形成能力测定来测定细胞存活率。通过流式细胞术测定 BPU 对细胞周期分布和细胞凋亡诱导的影响。根据单独或 BPU 后暴露于 X 射线的细胞的核伽玛 H2AX 免疫荧光分析 BPU 对辐射诱导的 DNA 损伤和修复的影响。使用流式细胞术测量抗-gamma H2AX 抗体染色（双链 DNA 断裂的替代决定因素）。结果 BPU (1.5 μM) 24 小时产生类似于 50% 的细胞存活率。 BPU 和 X 射线照射在三种细胞系中具有协同作用，存活水平为 20-50%。使用 BPU（1.5 μM，24 小时）重复实验的流式细胞术分析表明，BPU 阻断了 S 和/或 G(2)/M 的细胞进展。与对照细胞相比，BPU 处理的细胞凋亡发生率范围从相似的 0.3% 到相似的 8%。经过 X 射线照射 24 小时后，用 BPU 预处理并在药物暴露后进行 X 射线照射的细胞显示出 γ H2AX 水平大约比仅暴露于 X 射线的细胞高出两倍。 结论 该研究确定 BPU 是一种新型放射增敏剂。作为 DNA 双链断裂替代标记的磷酸化组蛋白 H2AX 的分析表明，放射增敏与 BPU 抑制 X 射线照射诱导的 DNA 损伤修复之间存在正相关。
Purpose The present study was designed to investigate the ability of N-[4-(5-bromo-2-pyrimidyloxy)-3-methylphenyl]-(dimemethylamino)-benzoylphenylurea (dimemethylamino benzoylphenylurea; BPU) to sensitize cells to radiation and to examine the relationship between phenotype versus survival, DNA damage, apoptosis, or cell cycle progression in non-small cell lung cancer (NSCLC) cell lines.Methods Asynchronous cultures of three NSCLC (phenotype) lines, A549 (adenocarcinoma), NCI-H226 (squamous) and NCI-H596 (adenosquamous) were used. Cells were treated for 24 h with BPU at various concentrations (0-10 mu M) to obtain drug doses for inhibiting cell survival by similar to 50% (IC50). Cells were X-irradiated without BPU or after 24 h BPU treatment at IC50. Radiation doses ranged from 0 to10 Gy. Cell survival was determined by a colony-forming ability assay. The effect of BPU on the cell cycle distribution and induction of apoptosis were measured by flow cytometry-based assays. The effect of BPU on radiation-induced DNA damage and repair was analyzed according to nuclear gamma H2AX immunofluorescence of cells exposed to X-rays alone or after BPU. Anti-gamma H2AX antibody staining, a surrogate determinant of double stranded DNA breaks, was measured using flow cytometry.Results BPU (1.5 mu M) for 24 h produced similar to 50% cell survival. BPU and X-irradiation were synergistic in the three cell lines at survival levels of 20-50%. Flow cytometry analysis of replicate experiments with BPU (1.5 mu M for 24 h) showed that BPU blocked cell progression at S and/or G(2)/M. The incidence of apoptosis in BPU-treated versus control cells ranged from similar to 0.3 to similar to 8%. Twenty-four hour after X-irradiation cells pre-treated with BPU and X-irradiated after drug exposure showed gamma H2AX levels approximately two times higher than did the cells exposed to X-rays only.Conclusions The study identified BPU as a novel radiation sensitizer. The analysis of phosphorylated histone H2AX as a surrogate marker of DNA double strand breaks suggested a positive association between radiosensitization and the inhibition of X-irradiation-induced DNA damage repair by BPU.