| 庄喜兵,乔田奎,许国雄,陈学,袁素娟,张琪,邢娜,陈伟.放射抗拒Lewis肺癌细胞系的建立[J].中华放射医学与防护杂志,2015,35(11):805-808 |
| 放射抗拒Lewis肺癌细胞系的建立 |
| Establishment of a radioresistant cell strain of Lewis lung cancer cells |
| 投稿时间：2015-03-31 |
| DOI：10.3760/cma.j.issn.0254-5098.2015.11.002 |
| 中文关键词: Lewis肺癌瘤系 电离辐射 放射抗拒 肿瘤免疫 |
| 英文关键词:Lewis lung cancer cell line Ionizing radiation Radioresistant Tumor immunity |
| 基金项目:上海市金山区卫计委面上项目(JSKJ-KTMS-2014-02) |
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| 中文摘要: |
| 目的 通过X射线反复照射体外培养的Lewis肺癌细胞系(LLC),建立其放射抗拒细胞系(R-LLC),并比较两种细胞系间Survivin蛋白的表达及分泌细胞因子水平的差异。方法 应用6MV X射线反复照射Lewis肺癌细胞系,5 Gy/次,共6次,30 Gy。采用CCK-8实验及克隆形成实验测定两种细胞系的放射敏感性;Western blot法检测两种细胞系Survivin蛋白表达的差异;ELISA法检测两种细胞系培养液上清细胞因子TNF-α、IL-12的浓度。结果 R-LLC及LLC细胞系的平均致死剂量D0分别是(1.397±0.128)和(1.053±0.214)Gy,外推值N分别是2.680±1.179和4.149±0.785,Dq值分别为1.038和1.988 Gy,SF2值分别为0.353±0.092和0.682±0.092,显示了明显的辐射抗性;Western blot结果显示,R-LLC细胞株的Survivin蛋白表达水平较LLC细胞株升高,差异有统计学意义(t=15.310,P<0.05);ELISA结果显示,R-LLC细胞上清液TNF-ɑ和IL-12浓度较LLC细胞明显升高,差异有统计学意义(t=2.867和3.014,P<0.05)。结论 反复照射使肿瘤细胞获得了辐射抗性,同时又促进了Lewis肺癌细胞Survivin蛋白及细胞因子的表达,为放射增敏及肿瘤免疫相关的研究提供依据。 |
| 英文摘要: |
| Objective To establish a radioresistant Lewis lung cancer cell line(R-LLC) by irradiating LLC cells with X-rays and to compare the expression levels of survivin and secreted cytokines between these two cell lines.Methods The Lewis lung cancer cells were irradiated with a total of 30 Gy of 6 MV X-rays from 6 irradiation periods of 5 Gy each. The radioresistance of LLC and R-LLC cells were confirmed by the CCK-8 and clonogenic assays;the expression level of Survivin was examined by Western blot assay and the concentrations of TNF-α and IL-12 in the culture supernatants were detected by ELISA. Results For the LLC and R-LLC cell lines, the average lethal doses D0 were(1.053±0.214) Gy and(1.397±0.128) Gy, the extrapolation values of N were(2.680±1.179) and(4.149±0.785), Dq equaled 1.038 and 1.988 Gy, and SF2 equaled 0.353±0.092 and 0.682±0.092, respectively, which demonstrated a significant radioresistance of R-LLC. The expression level of Survivin in R-LLC cells was significantly higher than that in LLC cells(t=15.310,P<0.05). ELISA assay also showed that the concentrations of TNF-ɑ and IL-12 in R-LLC cell culture supernatants were significantly higher than that in LLC cell culture supernatants(t=2.867, 3.014, P<0.05). Conclusions Cancer cell radiosensitivity is enhanced by multi-step irradiation together with higher expressions of survivin and cytokines in LLC cells. |
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