靶向survivin的siRNA联合凋亡相关配体对肝癌细胞的影响  

siRNA targeting survivin induces sensitization of hepatocarcinoma cells to tumor necrosis factor related apoptosis-inducing ligand

作　　者：卢昕[1] 郑启昌[1] 舒晓刚[1] 熊俊[1] 秦涛[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院肝胆外科,武汉430022

出　　处：《中华普通外科杂志》2006年第7期523-525,共3页Chinese Journal of General Surgery

摘　　要：目的探讨靶向survivin的siRNA联合肿瘤坏死因子诱导凋亡相关配体(tumor necrosis factor related apoptosis-inducing ligand,TRAIL)对肝癌细胞增殖与凋亡的影响。方法构建靶向survivin基因的siRNA真核表达载体,观察可溶性TRAIL对肝癌细胞增殖与凋亡的影响。结果采用RT-PCR检测显示,siRNA在mRNA水平抑制survivin基因表达为73％。MTT法检测到可溶性TRAIL对HepG2、HepG2／Silence(-)细胞增殖无明显的抑制作用(P>0．05),通过抑制survivin表达,可使HepG2／Silence(+)细胞的12 h存活率明显降低,48 h降至最低[HepG2／Silence(+)0．518±0．017,对照组0．741±0．005,P<0．01]。荧光显微镜检测HepG2／Silence(+)细胞12 h、24 h、48 h的细胞凋亡率分别为:11．85％±0．72％、28．97％±0．43％、41．80％±0．90％,与对照组相比差异有统计学意义(F=22．37,P<0．05)。结论靶向survivin的siRNA在抑制肝癌细胞中survivin表达的同时可增强TRAIL作用的敏感性。Objective To study the effect of siRNA targeting survivin and tumor necrosis factor related apeptosis-inducing ligand on the proliferation and apeptosis of hepatocellular carcinoma （HCC） cells. Methods siRNA eukaryotic expression vector was constructed and the effects of soluble tumor necrosis factor related apoptosis-inducing ligand （sTRAIL）on HCC cells were observed. Results The recombinant plasmid Psilence （ ± ）-survivin was successfully constructed. Survivin mRNA expression inhibition ratio reached 73% by RT-PCR. Observed by MTY method sTRAIL failed to inhibit the survival rate of HepG2,HepG2/Silence（ - ） cells at 12 h,24 h ,48 h when compared to control groups. With the survivin gene being inhibited, the survival rate of HepG2/Silence（ ± ） cells（0. 518 ±0. 017） decreased in 12 h compared to control groups （0. 741 ± 0. 005 ） and reached the lowest level in 48 h （ P 〈 0. 05 ）. Significant apeptosis was observed by fluorescence microscopy. The apoptosis ratio of HepG2/Silence （ ＋ ） cells in 12 h,24 h,48 h was （ 11.85 ± 0. 72） %, （28. 97 ± 0. 43 ） %, （41.80 ± 0. 90） % respectively and the difference was significant when compared with control group （ all P 〈 0. 05 ）. Conclusion siRNA targeting survivin suppresses the expression of survivin in HCC cells, and enhances the sensitivity of target HCC cell lines to sTRAIL.

关 键 词：癌 肝细胞 细胞凋亡 基因 肿瘤坏死因子诱导凋亡相关配体 

分 类 号：R735.7[医药卫生—肿瘤]