| 刘翠花,董新宇,李远航,张新强,王志成,赵刚,申延男.解耦联蛋白2对Siha细胞辐射敏感性的影响[J].中华放射医学与防护杂志,2020,40(2):88-93 |
| 解耦联蛋白2对Siha细胞辐射敏感性的影响 |
| Effect of uncoupling protein 2 gene on radiation sensitivity of Siha cells |
| 投稿时间：2019-04-05 |
| DOI：10.3760/cma.j.issn.0254-5098.2020.02.003 |
| 中文关键词: 宫颈癌 RNA干扰 解耦联蛋白2 细胞凋亡 辐射敏感性 |
| 英文关键词:Cervical cancer RNA interference Uncoupling protein 2 Apoptosis Radiation sensitivity |
| 基金项目:国家自然科学基金（31600678） |
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| 中文摘要: |
| 目的 探究沉默解耦联蛋白2（UCP2）联合辐照对Siha细胞辐射敏感性的影响，为宫颈癌放疗增敏提供新的靶点。方法 将UCP2 siRNA转染入Siha细胞后进行X射线照射，通过集落形成、CCK-8、流式细胞实验来验证UCP2对Siha细胞辐射敏感性作用，检测线粒体膜电位和活性氧（ROS）的产生来进一步探讨相关机制。结果 RT-PCR和Western blot表明Siha细胞经辐照后UCP2表达增加。成功构建UCP2沉默模型，沉默组（siUCP2）D0、Dq、N、SF2分别为1.54、1.31、2.31 Gy和0.52，空白对照组（mock）分别为2.50、3.64、4.30 Gy和0.83，阴性对照组（siNC）分别为3.34、2.16、1.91 Gy和0.69，沉默组较空白对照组和阴性对照组放射增敏比分别为0.62和0.46，并且沉默组细胞增殖活性较对照组明显降低（t=13.2，P<0.05）；辐照后沉默组细胞凋亡水平显著高于对照组（t=3.14，P<0.05）；照射后12 h，沉默组的ROS产量明显高于对照组（t=19.10，P<0.05）；照射后24 h，沉默组的Siha细胞线粒体膜电位较对照组显著降低（t=8.87，P<0.05）。结论 UCP2沉默后的Siha细胞辐射敏感性增强，并且可能会成为宫颈癌细胞辐射增敏的新靶点。 |
| 英文摘要: |
| Objective To investigate whether silencing UCP2 can sensitize cervical cancer cell line Siha to radiation. Methods Siha cells were transfected with UCP2 siRNA and then irradiated by X-ray. The radiosensitivity of Siha cells was verified by colony formation, CCK-8, apoptosis and immunofluorescence assays. The mitochondrial membrane potential and the production of reactive oxygen species (ROS) were detected to further explore the related mechanism. Results RT-PCR and Western blot assays showed that the expression of UCP2 in Siha cells was increased after irradiation and the UCP2 siRNA successfully silenced the expression of UCP in cells. According to the survival curves, the D0, Dq, N and SF2 were 1.54, 1.31, 2.31 Gy and 0.52 for siUCP2 group, 2.50, 3.64, 4.30 Gy and 0.83 for blank control group, and 3.34, 2.16, 1.91 Gy and 0.69, for siNC group, respectively. The radiosensitivity enhancement ratio of silent group was 0.62 and 0.46, compared with blank control group and negative control group, respectively. The proliferative activity of cells in the silent group was lower than that in the control group (t=13.2, P<0.05). Apoptosis levels in the silent group were significantly higher than those in the control group after irradiation(t=3.14,P<0.05). At 4 h after irradiation, the ROS production in the silent group was significantly higher than that in the control group (t=19.10, P<0.05). At 24 h after irradiation, the mitochondrial membrane potential of Siha cells in the silent group was significantly lower than that in the control group (t=4.18, P<0.05). Conclusions The radiosensitivity of Siha cells is enhanced after UCP2 silencing, and thus UCP2 may applicable as a new target for radiosensitization of cervical cancer cells. |
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