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利用重离子加速器国家重点实验室提供的12C6+束流照射人宫颈瘤(HeLa)细胞,测量辐射结束后细胞克隆率绘制细胞存活曲线。使用10μg/mL的维拉帕米处理细胞作为一组对照,检测维拉帕米对于HeLa细胞辐射敏感性的影响。利用Fluo-3 AM钙离子荧光探针配合流式细胞仪检测重离子照射后的胞内游离钙(IECa2+)水平。实验结果表明0～5 Gy范围内重离子辐射剂量与HeLa细胞存活率负相关,照射结束后胞质游离钙浓度有一定程度的上升。而维拉帕米能显著降低HeLa细胞经重离子辐射后的存活及增殖能力,且胞质游离钙浓度处于较低水平。造成这种现象的原因可能是维拉帕米能降低细胞对非致死性DNA损伤的修复能力。
The HeLa cells were irradiated with 12C6 + beam provided by the State Key Laboratory of Heavy Ion Accelerator, and the cell survival curve was drawn after the radiation was finished. Cells were treated with verapamil at 10 μg / mL as a set of controls to examine the effect of verapamil on HeLa cell radiation sensitivity. The level of intracellular free calcium (IECa2 +) after heavy ion irradiation was detected by Fluo-3 AM calcium ion fluorescence probe combined with flow cytometry. The experimental results showed that the dose of heavy ion in the range of 0-5 Gy was negatively correlated with the survival rate of HeLa cells. After the irradiation, the concentration of cytosolic free calcium increased to a certain extent. However, verapamil significantly reduced the viability and proliferation of HeLa cells after exposure to heavy-ion radiation, and the cytosolic free calcium concentration was at a low level. The reason for this phenomenon may be verapamil can reduce the cell repair of non-lethal DNA damage.