中华放射医学与防护杂志
中華放射醫學與防護雜誌
중화방사의학여방호잡지
Chinese Journal of Radiological Medicine and Protection
2012年
1期
40-43,59
,共5页
bcl-2%XIAP%UMSCC12%放射增敏%凋亡
bcl-2%XIAP%UMSCC12%放射增敏%凋亡
bcl-2%XIAP%UMSCC12%방사증민%조망
bcl-2%XIAP%UMSCC12%Radiosensitization%Apoptosis
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(f=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.
目的 探討同時沉默bcl-2和XIAP基因對頭頸部鱗癌UMSCC12細胞放射敏感性的影響.方法 實驗分為4組:對照siRNA轉染組、siRNA-bcl-2轉染組、siRNA-XIAP轉染組和siRNA-bcl-2與siRNA-XIAP共轉染組.採用siRNA bcl-2和siRNA-XIAP共轉染頭頸鱗癌細胞株UMSCC12,Western blot檢測蛋白水平基因沉默的效果.以caspase-3和caspase-9活性檢測自髮和放療誘導的凋亡,最後以剋隆形成實驗評價放射增敏效果.結果 共轉染siRNA-bcl-2和siRNA-XIAP有效沉默瞭UMSCC12細胞bcl-2和XIAP的蛋白錶達.caspase-3和caspase-9活性檢測提示,單獨沉默XIAP未增加細胞自髮和放射誘導的凋亡.單獨沉默bcl-2使放射誘導的凋亡增加,與對照siRNA轉染組照射後相比,caspase-3和caspase-9的活性差異有統計學意義(t=5.32、6.27,P＜0.05),但未增加細胞自髮的凋亡.共轉染後的caspase-3和caspase-9活性在未照射時比對照siRNA轉染組分彆提高至1.36和1.34倍(t=11.47、6.22,P＜0.05),照射後分彆提高至1.72和1.98倍(f=12.02、20.14,P＜0.05).剋隆形成實驗顯示,與對照siRNA轉染組比較,siRNA-XIAP的放射增敏比(SER)為1.06,siRNA-bcl-2的放射增敏比為1.15,siRNA-bcl-2與siRNA-XIAP共轉染組的放射增敏比為1.41,比其他組顯著升高.結論 與單獨沉默bcl-2和XIAP相比,同時沉默bcl-2和XIAP增加瞭UMSCC12細胞的放射敏感性,其機製可能與增加瞭UNSCC12細胞自髮和放射誘導的凋亡有關.
목적 탐토동시침묵bcl-2화XIAP기인대두경부린암UMSCC12세포방사민감성적영향.방법 실험분위4조:대조siRNA전염조、siRNA-bcl-2전염조、siRNA-XIAP전염조화siRNA-bcl-2여siRNA-XIAP공전염조.채용siRNA bcl-2화siRNA-XIAP공전염두경린암세포주UMSCC12,Western blot검측단백수평기인침묵적효과.이caspase-3화caspase-9활성검측자발화방료유도적조망,최후이극륭형성실험평개방사증민효과.결과 공전염siRNA-bcl-2화siRNA-XIAP유효침묵료UMSCC12세포bcl-2화XIAP적단백표체.caspase-3화caspase-9활성검측제시,단독침묵XIAP미증가세포자발화방사유도적조망.단독침묵bcl-2사방사유도적조망증가,여대조siRNA전염조조사후상비,caspase-3화caspase-9적활성차이유통계학의의(t=5.32、6.27,P＜0.05),단미증가세포자발적조망.공전염후적caspase-3화caspase-9활성재미조사시비대조siRNA전염조분별제고지1.36화1.34배(t=11.47、6.22,P＜0.05),조사후분별제고지1.72화1.98배(f=12.02、20.14,P＜0.05).극륭형성실험현시,여대조siRNA전염조비교,siRNA-XIAP적방사증민비(SER)위1.06,siRNA-bcl-2적방사증민비위1.15,siRNA-bcl-2여siRNA-XIAP공전염조적방사증민비위1.41,비기타조현저승고.결론 여단독침묵bcl-2화XIAP상비,동시침묵bcl-2화XIAP증가료UMSCC12세포적방사민감성,기궤제가능여증가료UNSCC12세포자발화방사유도적조망유관.
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 radiationinduced 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.
