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中华放射肿瘤学杂志
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中华放射肿瘤学杂志  2013, Vol. 22 Issue (4): 326-328    DOI: 10.3760/cma.j.issn.1004-4221.2014.04.019
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RNA干扰抑制UHRF1基因表达对食管癌细胞放射增敏作用的研究
杨从容, 王雅棣, 李成林, 景绍武, 孙国贵
100700 北京军区总医院放疗科
UHRF1 expression inhibition by RNA interference enhances the radiosensitivity of esophageal cancer cells
YANG Cong-rong*, WANG Ya-di, LI Cheng-lin, JING Shao-wu, SUN GUO-gui
Department of Radiation Oncology, Military General Hospital of Beijing, Beijing 100700, China
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摘要  目的 研究RNA干扰抑制UHRF1表达对食管癌细胞系(TE-1)放射敏感性的影响及作用机制。方法 以慢病毒感染方法将UHRF1基因的短发夹状RNA (shRNA)转入TE-1细胞,并分为未转染组、转染NC-shRNA组、转染UHRF1-shRNA组,前二者为对照组。采用 RT-PCR和蛋白印记法检测转染前后细胞UHRF1的mRNA和蛋白表达,成克隆法、流式细胞术及蛋白印记法检测转染UHRF1-shRNA联合X线照射对TE-1细胞放射敏感性、周期、凋亡及DNA损伤标识蛋白γ-H2AX的影响。
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杨从容
王雅棣
李成林
景绍武
孙国贵
关键词 UHRF1基因放射敏感性G2+M期阻滞凋亡DNA损伤修复    
Abstract:Objective To study the effect of UHRF1 expression inhibition by RNA interference on the radiosensitivity of esophageal cancer cell line TE-1 and its mechanism. Methods Short hairpin RNA (shRNA) targeting UHRF1 gene was introduced into TE-1 cells by lentivector-mediated transfer. The cells were divided into three groups:non-transfected group, negative control (NC)-shRNA-transfected group, and UHRF1-shRNA-transfected group. The mRNA and protein expression levels of UHRF1 in TE-1 cells were measured by RT-PCR and Western blot before and after transfection. After transfection and X-ray radiation, the radiosensitivity of TE-1 cells was evaluated by colony formation assay;the cell cycle and cell apoptosis were determined by flow cytometry;the γ-H2AX (as a marker of DNA damage) level was measured by Western blot. Results After transfection with UHRF1-shRNA, the mRNA and protein expression levels of UHRF1 were significantly decreased in TE-1 cells, as compared with those in the NC-shRNA-transfected group and non-transfected group (0.11 vs 0.96 and 0.98, F=124.21, P=0.000;0.10 vs 0.89 and 0.94, F=125.25, P=0.000). The UHRF1-shRNA-transfected group had sensitization enhancement ratios of 1.53(D0 ratio) and 1.95(Dq ratio). X-ray radiation could cause G2/M arrest and increase apoptotic rate and γ-H2AX expression in TE-1 cells. Compared with the two control groups, the UHRF1-shRNA-transfected group showed significantly less G2/M arrest (F=500.15, P=0.000), a significantly higher apoptotic rate (F=100.10,P=0.000), and significantly higher residual γ-H2AX expression (F=61.00,P=0.000) at 24 hours after X-ray radiation. Conclusions RNA interference can effectively inhibit the UHRF1 expression and enhance the radiosensitivity of TE-1 cells. The mechanism may be related to cell cycle regulation, cell apoptosis, and DNA damage repair.
Key words UHRF1 gene    Radiosensitivity    G2/M arrest    Apoptosis    DNA damage repair   
收稿日期: 2013-01-05     
通讯作者: 王雅棣     E-mail: wangyadi@hotmail.com
引用本文:   
杨从容,王雅棣,李成林等. RNA干扰抑制UHRF1基因表达对食管癌细胞放射增敏作用的研究[J]. 中华放射肿瘤学杂志, 2013, 22(4): 326-328.
YANG Cong-rong,WANG Ya-di,LI Cheng-lin et al. UHRF1 expression inhibition by RNA interference enhances the radiosensitivity of esophageal cancer cells[J]. Chinese Journal of Radiation Oncology, 2013, 22(4): 326-328.
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http://journal12.magtechjournal.com/Jweb_fszlx/CN/10.3760/cma.j.issn.1004-4221.2014.04.019     或     http://journal12.magtechjournal.com/Jweb_fszlx/CN/Y2013/V22/I4/326
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