中华放射医学与防护杂志

李文雁,刘巍,孟晓洁.siRNA共转染沉默bcl-2和XIAP基因对UMSCC12细胞放射增敏作用的实验研究[J].中华放射医学与防护杂志,2012,32(1):40-43,59.LI Wen-yan,LIU Wei,MENG Xiao-jie.Simultaneous silence of bcl-2 and XIAP increases radiosensitivity of UMSCC12 head and neck cancer cells[J].Chin J Radiol Med Prot,2012,32(1):40-43,59

siRNA共转染沉默bcl-2和XIAP基因对UMSCC12细胞放射增敏作用的实验研究

Simultaneous silence of bcl-2 and XIAP increases radiosensitivity of UMSCC12 head and neck cancer cells

投稿时间：2011-07-09

DOI：10.3760/cma.j.issn.0254-5098.2012.01.009

中文关键词: bcl-2 XIAP UMSCC12 放射增敏凋亡

英文关键词:bcl-2 XIAP UMSCC12 Radiosensitization Apoptosis

基金项目:

作者	单位
李文雁	050051 石家庄, 河北医科大学第三医院放疗科
刘巍	河北医科大学第四医院肿瘤内科
孟晓洁	Department of Molecular Biosciences and Urology, University of Kansas, Lawrence, KS, USA

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中文摘要:

目的探讨同时沉默bcl-2 和XIAP基因对头颈部鳞癌UMSCC12细胞放射敏感性的影响。方法实验分为4组:对照siRNA转染组、siRNA-bcl-2转染组、siRNA-XIAP转染组和siRNA-bcl-2与siRNA-XIAP共转染组。采用siRNA bcl-2和 siRNA-XIAP 共转染头颈鳞癌细胞株UMSCC12,Western blot检测蛋白水平基因沉默的效果。以caspase-3和caspase-9活性检测自发和放疗诱导的凋亡,最后以克隆形成实验评价放射增敏效果。结果共转染siRNA-bcl-2和siRNA-XIAP有效沉默了UMSCC12细胞bcl-2和XIAP的蛋白表达。caspase-3和caspase-9活性检测提示,单独沉默XIAP未增加细胞自发和放射诱导的凋亡。单独沉默bcl-2使放射诱导的凋亡增加,与对照siRNA转染组照射后相比,caspase-3 和caspase-9的活性差异有统计学意义(t=5.32、6.27,P<0.05),但未增加细胞自发的凋亡。共转染后的caspase-3和caspase-9活性在未照射时比对照siRNA转染组分别提高至1.36和1.34倍(t=11.47、6.22,P<0.05),照射后分别提高至1.72和1.98倍(t=12.02、20.14,P<0.05)。克隆形成实验显示,与对照siRNA转染组比较,siRNA-XIAP 的放射增敏比(SER)为1.06,siRNA-bcl-2 的放射增敏比为1.15,siRNA-bcl-2与siRNA-XIAP共转染组的放射增敏比为1.41,比其他组显著升高。结论与单独沉默bcl-2和XIAP相比,同时沉默bcl-2和XIAP增加了UMSCC12细胞的放射敏感性,其机制可能与增加了UNSCC12细胞自发和放射诱导的凋亡有关。

英文摘要:

Objective To investigate the radiosensitization effect of the simultaneous silence of bcl-2 and XIAP on human head and neck cancer cell line in vitro. Methods Four groups of UMSCC12 cells were transfected with siRNA-bcl-2, siRNA-XIAP, siRNA-bcl-2 plus siRNA XIAP, and siRNA control, respectively. The silence efficiency was tested by Western blot assay. Apoptosis was evaluated with the activities of caspase-3 and caspase-9, and radiosensitization effect was evaluated with clonogenic assay. Results Bcl-2 and XIAP protein expressions were effectively eliminated in the cells simultaneously transfected with siRNA-bcl-2 and siRNA-XIAP. Transfection of cells with bcl-2 siRNA increased radiation-induced apoptosis (t=5.32,6.27;P<0.05), but transfection with XIAP siRNA did not impact cell apoptosis. Since the simultaneous transfection of the above two siRNAs, the activities of caspase-3 and caspase-9 were increased by 1.36 and 1.34 times (t=11.47,6.22;P<0.05)in nonirradiated cells and increased by 1.72 and 1.98 times (t=12.02,20.14;P<0.05) in irradiated cells, respectively. The colonic assay showed that the SERs were 1.06, 1.15 and 1.41 for the cells transfected with XIAP siRNA, bcl-2 siRNA and both siRNAs, respectively. Conclusions Compared to single silence of XIAP or bcl-2, simultaneous silence of XIAP and bcl-2 offers a potential approach to improving the radiosensitivity of the head and neck cancer cells, and apoptosis might contribute to the enhancement of radiosensitization.

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