董娟聪,张海芹,高辉,等.神经纤毛蛋白1对非小细胞肺癌放射敏感性的影响[J].中华放射医学与防护杂志,2014,34(11):809-812.Dong Juancong,Zhang Haiqin,Gao Hui,et al.Effect of neuropilin-1 on radio-sensitivity of non-small cell lung carcinoma[J].Chin J Radiol Med Prot,2014,34(11):809-812 |
神经纤毛蛋白1对非小细胞肺癌放射敏感性的影响 |
Effect of neuropilin-1 on radio-sensitivity of non-small cell lung carcinoma |
投稿时间:2014-04-25 |
DOI:10.3760/cma.j.issn.0254-5098.2014.11.003 |
中文关键词: 神经纤毛蛋白1 非小细胞肺癌 A549细胞 放射敏感性 辐射抗性 |
英文关键词:Neuropilin 1 Non-small cell lung carcinoma A549 cell Radiosensitivity Radioresistance |
基金项目:国家自然科学基金(30870584,81371890);教育部高等学校博士学科点专项科研基金(20120061110063) |
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中文摘要: |
目的 探讨神经纤毛蛋白1 (neuropilin 1,NRP1)在不同种类非小细胞肺癌(NSCLC)细胞系的表达情况及其与肿瘤放射敏感性的关系.方法 通过Western blot技术检测不同来源的5种人NSCLC细胞系(H358、H460、H1299、A549、SK-MES-1)NRP1的基础表达水平,MTT方法检测经不同剂量X射线照射后肺癌细胞的存活情况,初步筛选5种细胞中辐射敏感和辐射抵抗的2种细胞;采用克隆形成实验及流式细胞术分别检测细胞存活分数及凋亡率的变化,分析2种肺癌细胞的放射敏感性,采用Western blot检测2种细胞受X射线作用后 NRP1 的表达变化,通过克隆形成实验检测靶向抑制NRP1对A549细胞放射敏感性的影响.结果 5种肿瘤细胞NRP1表达量由高到低依次为:A549>SK-MES-1>H358>H460>H1299;X射线对肿瘤细胞的抑制能力呈剂量依赖性增强,其抑制率由高到低依次为:H460>H1299>H358>SK-MES-1>A549,因此选取A549细胞和 H460细胞做后续研究.克隆形成实验显示,A549在2 Gy照射下的存活分数(SF2)是0.887,而H460细胞为0.313.A549细胞受照后的凋亡率为对照组的122.54%,明显低于H460细胞的238.88%.辐射可以诱导A549细胞NRP1的表达上调达40%,同时靶向抑制NRP1则可以增强A549细胞的放射敏感性.结论 在NSCLC细胞中A549细胞具有较强的辐射抗性,且辐射抗性的形成与其NRP1表达上调有关. |
英文摘要: |
Objective To investigate the expression and relationship with radiosensitivity of neuropilin-1 (NRP-1) of non-small cell lung carcinoma (NSCLC) cells. Methods The basal expression level of NRP1 in the cells of H358, H460, H1299, A549 and SK-MES-1were detected by Western blot. MTT method was used to measure the apoptotic rate, and two kinds of cells were selected for further study on radiosensitivity and radioresistance. The surviving fraction (SF2) and apoptosis rate were analyzed with clonogenic assay and flow cytometry respectively. Results The expression level from high to low of NRP1 in order was: A549>SK-MES-1>H358>H460>H1299, while the radiosensitivity from strong to weak was H460>H1299>H358>SK-MES-1>A549. Therefore, A549 and H460 cells were chosen to do follow-up studies. The SF was 0.887 at 2 Gy in A549 cells,while 0.313 in H460 cells. After 10 Gy X-rays, the apoptosis rate of H460 cells was 238.88%, significantly higher than that of A549 cells (122.54%). Ionizing radiation induced up-regulation of NRP1 expression in A549 cells by up to 40%, while the targeted inhibition of NRP1 enhanced the radiosensitivity of A549 cells. Conclusions A549 cells were more radioresistance than other cells, which may be associated with the up-regulation of NRP1 expression. |
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