| 孔飞飞,李元杰,马璐,隋御,折虹,徐方.沉默Rev1基因对人结肠癌细胞放射敏感性的影响[J].中华放射医学与防护杂志,2018,38(10):721-727 |
| 沉默Rev1基因对人结肠癌细胞放射敏感性的影响 |
| Effect of Rev1 siRNA on the sensitivity of human colon cancer cells to X-ray radiation |
| 投稿时间：2018-06-04 |
| DOI：10.3760/cma.j.issn.0254-5098.2018.10.001 |
| 中文关键词: 结肠癌细胞 Rev1基因 X射线 放射敏感性 |
| 英文关键词:Colon cancer cells Rev1 X-rays Radiosensitivity |
| 基金项目:国家自然科学基金（31360251）；宁夏回族自治区重点研发计划项目（宁科技字[2016]25号） |
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| 中文摘要: |
| 目的 探讨沉默Rev1基因对人高分化结肠癌细胞THC8307 X射线照射后增殖、凋亡的影响。方法 利用RNA干扰技术将Rev1基因的特异性片段转染THC8307细胞，用实时荧光定量PCR和Western blot检测转染后细胞中目的基因的表达；实验分空白对照组、阴性对照组、Rev1 siRNA组。分别给予0、6 Gy X射线照射，运用四甲基偶氮唑盐（MTT）法检测不同时段（0、24、48、72、96 h）细胞增殖情况，并绘制增殖曲线；流式细胞仪（FCM）检测不同剂量X射线处理后的细胞凋亡；Western blot检测各组PCNA、γ-H2AX、P53、Bax、Bcl-2蛋白的表达。结果 与空白对照组相比，6 Gy X射线照射后，沉默Rev1基因组细胞增殖速率明显降低（t=7.53，P<0.05），凋亡明显增加（t=6.23，P<0.05），增殖细胞抗原（PCNA）表达下降（t=4.39，P<0.05），γ-H2AX表达升高（t=5.48，P<0.05），P53表达升高（t=5.09，P<0.05）、Bax表达升高（t=3.32，P<0.05）、Bcl-2表达降低（t=6.13，P<0.05）。结论 沉默Rev1基因，抑制了X射线作用下的结肠癌细胞THC8307的增殖，促进了凋亡，增加了其对X射线的放射敏感性。 |
| 英文摘要: |
| Objective To explore the effect of silencing Rev1 gene on the proliferation and apoptosis induction of human colon cancer cell line THC8307 after X-ray irradiation. Methods Rev1 siRNA was transferred into THC8307 cells and its transferring efficiency was tested by RT-PCR and Western blot assay. The cells were divided into blank control group, negative control group and radiation group. After 6 Gy irradiation, cell proliferation were detected by MTT assay, cell apoptosis were detected by flow cytometry (FCM), and the expressions of PCNA, γ-H2AX, P53, Bax, and Bcl-2 proteins in each group were detected by Western blot. Results Compared with the blank control group, after 6 Gy irradiation, the proliferation rate of the silenced Rev1 was reduced (t=7.53, P<0.05) and apoptosis rate was increased (t=6.23, P<0.05). The expression of PCNA and Bcl-2 decreased (t=4.39, 6.13 P<0.05), and the expressions of γ-H2AX, P53 and Bax increased (t=5.48, 5.09, 3.32, P<0.05). Conclusions Silencing Rev1 gene inhibits the proliferation of colon cancer cells, promotes apoptosis, and increases cell radiosensitivity to X-rays. |
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