| 黄培,钱红英,孙屏,张熔熔,赵帆,陈义钢.DDX46基因对结肠癌细胞电离辐射敏感性的影响及机制研究[J].中华放射医学与防护杂志,2018,38(4):247-252 |
| DDX46基因对结肠癌细胞电离辐射敏感性的影响及机制研究 |
| Effects of DDX46 gene on the radiosensitivity of colorectal cancer cells |
| 投稿时间：2017-11-09 |
| DOI：10.3760/cma.j.issn.0254-5098.2018.04.002 |
| 中文关键词: 结直肠癌 DDX46 辐射敏感性 |
| 英文关键词:Colorectal cancer cells (CRC) DDX46 Radiosensitivity |
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| 中文摘要: |
| 目的 探讨DDX46基因对结肠癌细胞的电离辐射敏感性的影响及其作用机制。方法 DDX46基因RNA干扰慢病毒转染的SW480细胞为实验组，空载质粒慢病毒转染的SW480细胞为对照组。两组细胞转染72 h后，分别进行0和4 Gy X射线照射，采用CCK-8方法检测两组的细胞活力；实验组联合4 Gy X射线照射24 h后，采用免疫荧光技术和Western blot法检测细胞γ-H2AX foci数量以及DNA损伤修复通路关键蛋白的表达水平，确定DDX46与辐射诱导的DNA损伤修复的关系。结果 4 Gy X射线照射24 h后，实验组的细胞活力比照射前降低（15.02±3.92）%（t=-4.696，P<0.05），比对照组降低（17.43±1.83）%（t=4.844，P<0.01）；而对照组照射前后比较差异无统计学意义（P>0.05）。同时，实验组γ-H2AX foci数量相比于对照组增加了（43.03±17.6）%（t=-3.108，P<0.05），ATM的蛋白水平显著升高（t=7.530，P<0.01），而ATM的活性形式p-ATM以及ATM的下游靶蛋白Rad50的蛋白水平则明显降低（t=4.260、4.260、P<0.05），同时γ-H2AX的蛋白表达水平也有一定程度的升高（t=-3.090，P<0.05），DNA-PK在两组细胞内的蛋白水平变化不大。结论 沉默DDX46可以提高结肠癌细胞系SW480的辐射敏感性，其机制可能是DDX46表达沉默抑制结肠癌细胞系SW480中ATM的活化，从而抑制辐射诱导的DNA损伤的修复，提高SW480的辐射敏感性。 |
| 英文摘要: |
| Objective To investigate the effect of DDX46 gene on the radiosensitivity of colorectal cancer cells and underlying mechanism. Methods SW480 cells transfected with DDX46 RNAi or its empty plasmid by lentivirus were set as experimental group and control group, respectively. After 72 h of transfection, the cells were irradiated with 4 Gy X rays. The cell viabilities of these two groups were detected by CCK-8 assay. The number of γ-H2AX foci and the expressions of some key proteins related to DNA damage repair were detected by immunofluorescence technique and Western blot at 24 h after irradiation. Results At 24 h after 4 Gy irradiation, the cell vitality of experimental group was decreased to(15.02±3.92)%(t=-4.696, P<0.05)of control and(17.43±1.83)%(t=4.844, P<0.01) of nonirradiated cells, but there was no significant difference between 4 Gy irradiated control cells and nonirradiated cells. Meanwhile, compared with the control group, the ATM protein expression level (t=7.530, P<0.01) and the number of γ-H2AX foci (t=-3.108, P<0.05) were significantly increased in the experimental SW480 cells,while the expression levels of p-ATM and Rad50 were significantly reduced (t=4.260, 4.260, P<0.05), and the protein levels of DNA-PK in these two groups had tiny difference. Conclusions DDX46 RNA silence increases the radiation sensitivity of SW480 cells by inhibiting ATM activation and DNA repair. |
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