| 殷丽娜,张旭霞,王晶,等.氯硝柳胺对人三阴性乳腺癌MDA-MB-231细胞的放射增敏作用研究[J].中华放射医学与防护杂志,2014,34(4):244-249.Yin Lina,Zhang Xuxia,Wang Jing,et al.Radiosensitization of human triple-negative breast cancer MDA-MB-231 cells by antihelminthic niclosamide[J].Chin J Radiol Med Prot,2014,34(4):244-249 |
| 氯硝柳胺对人三阴性乳腺癌MDA-MB-231细胞的放射增敏作用研究 |
| Radiosensitization of human triple-negative breast cancer MDA-MB-231 cells by antihelminthic niclosamide |
| 投稿时间：2013-08-27 |
| DOI：10.3760/cma.j.issn.0254-5098.2014.04.002 |
| 中文关键词: 氯硝柳胺 放射增敏 β-连环蛋白 MDA-MB-231细胞 |
| 英文关键词:Niclosamide Radiosensitization β-catenin MDA-MB-231 cells |
| 基金项目:上海市自然科学基金（13ZR1403500） |
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| 中文摘要: |
| 目的 研究氯硝柳胺对人三阴性乳腺癌MDA-MB-231细胞的放射增敏作用及其与Wnt/β-连环蛋白信号通路相关的作用机制。方法 四甲基偶氮唑蓝（MTT）法检测不同浓度的氯硝柳胺对MDA-MB-231细胞生长的抑制效应，求得IC50值。将细胞分为空白对照组、氯硝柳胺单纯给药组、单纯照射组和氯硝柳胺+照射组，氯硝柳胺预处理的浓度为1.0和1.5 μmol/L，137Cs γ射线的照射剂量为0、2、4和6 Gy；克隆形成试验检测氯硝柳胺对MDA-MB-231细胞的放射增敏作用；免疫荧光法检测辐射诱导的γH2AX 焦点形成；流式细胞仪检测细胞周期的变化；Western blot法检测磷酸化和非磷酸化β-连环蛋白和Cyclin D1蛋白表达。结果 氯硝柳胺抑制MDA-MB-231细胞生长24 h的IC50值为13.63 μmol/L；1.0和1.5 μmol/L氯硝柳胺对MDA-MB-231细胞均有较好的放射增敏作用，放射增敏比SER分别为1.37和1.62，随给药剂量的增大而增强；氯硝柳胺预处理使受照MDA-MB-231细胞的γH2AX焦点形成率较单纯照射组显著增加（t=3.91， P<0.05），G2/M期阻滞明显减少（t=8.05，P<0.01），并显著抑制了辐射诱导的磷酸化β-连环蛋白（S675）、非磷酸化β-连环蛋白和Cyclin D1蛋白表达的升高。结论 氯硝柳胺对MDA-MB-231细胞具有显著的放射增敏作用，其作用机制与抑制Wnt/β-连环蛋白信号通路，从而抑制DNA DSBs修复和减少辐射诱导的G2/M期阻滞有关。Wnt/β-连环蛋白信号通路可能是有效的三阴性乳腺癌放射增敏的新靶点。 |
| 英文摘要: |
| Objective To investigate the radiosensitization effect of antihelminthic niclosamide on human triple-negative breast cancer MDA-MB-231 cells and the potential mechanism related to Wnt/β-catenin signaling pathway. Methods Four methyl thiazolyl tetrazolium(MTT)assay was used to measure the effect of niclosamide on cell viability at different concentrations and 50% inhibitory concentration(IC50)value was calculated. MDA-MB-231 cells were divided into 4 groups: untreated control, niclosamide treatment alone group, radiation alone group and niclosamide plus radiation treatment group. The cells with or without 1.0 and 1.5 μmol/L niclosamide pre-treatment were irradiated with 137Cs γ-rays at doses of 0, 2, 4 and 6 Gy. Cell survival was assayed with the colony formation method, radiation-induced γH2AX foci was analyzed with immunofluorescence, cell cycle progression was assayed with flow cytometry, and the changes of phospho-and non-phospho-β-catenin and Cyclin D1 protein expressions were measured with Western blot. Results Niclosamde obviously inhibited the viability of MDA-MB-231 cells in a dose-dependent manner with a IC50 value of 13.63 μmol/L. Pretreatment of cells with 1.0 and 1.5 μmol/L niclosamide evidently enhanced the radiosensitivity of MDA-MB-231 cells to γ-rays, and the values of SER were 1.37 and 1.62, respectively. Niclosamide pretreatment significantly increased radiation-induced γH2AX foci formation(t=3.91, P<0.05), diminished the radiation-induced G2/M arrest(t=8.05, P<0.01), and inhibited radiation-induced expressions of phospho-β-catenin (S675),non-phospho-β-catenin and Cyclin D1 proteins in MDA-MB-231 cells. Conclusions Niclosamide significantly can enhance the sensitivity of MDA-MB-231 cells to γ-ray irradiation through inhibiting Wnt/β-catenin signaling pathway, which results in the inhibition of DNA DSBs repair and the reduction of radiation-induced G2/M arrest. Wnt/β-catenin signaling pathway may serve as an ideal molecular target for radiosensitization of triple-negative breast cancer. |
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