沉默Rev1基因对人结肠癌细胞放射敏感性的影响  

Effect of Revl siRNA on the sensitivity of human colon cancer cells to X-ray radiation

作　　者：孔飞飞 李元杰[1] 马璐[1] 隋御[1] 折虹[2] 徐方[1] Kong Feifei;Li Yuanjie;Ma Lu;Sui Yu;Zhe Hong;Xu Fang(Key Laboratory of Fertility Preservation and Maintenance of the Ministry of Education,Ningxia Medical University,Yinchuan 750004,Chin;Department of Radiotherapy,Cancer Hospital,General Hospital of Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学生育力保持教育部重点实验室,银川750004 [2]宁夏医科大学总医院附属肿瘤医院放疗科,银川750004

出　　处：《中华放射医学与防护杂志》2018年第10期721-727,共7页Chinese Journal of Radiological Medicine and Protection

基　　金：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）

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。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.

关 键 词：结肠癌细胞 Rev1基因 X射线 放射敏感性 

分 类 号：R735.35[医药卫生—肿瘤]