应用RNA干扰技术阻断SYT-SSX表达治疗滑膜肉瘤的体外实验研究  

Knockdown fusion gene SYT-SSX expression by RNAi induces apoptosis in synovial sarcoma cells line HS-SY-Ⅱ

作　　者：杨毅[1] 郭卫[1] 李大森[1] 

机构地区：[1]北京大学人民医院骨与软组织肿瘤治疗中心,100044

出　　处：《中国骨肿瘤骨病》2006年第3期142-146,共5页Chinse Journal Of Bone Tumor And Bone Disease

基　　金：国家自然基金资助项目(编号:30571869/c03030306)

摘　　要：目的研究siRNA(smallinterferingRNA)对滑膜肉瘤细胞系HS-SY-II中融合基因SYT-SSX表达的抑制作用,为RNAi技术在肿瘤生物治疗中的应用提供实验基础。方法体外合成3条针对滑膜肉瘤特异性融合基因SYT-SSX的siRNA,转染HS-SY-II细胞系,分别应用逆转录聚合酶链反应(RT-PCR)和Western杂交方法检测SYT-SSX基因和蛋白的表达情况,并用MTT比色法测定细胞的增殖活性,Annexin-FITC/PI双标记流式细胞计数检测细胞凋亡。结果设计的siRNA中,1条可以完全封闭SYT-SSX的转录,另2条分别使SYT-SSX基因表达降低至37.3%和70.1%。转染3条siRNA后,HS-SY-II细胞融合蛋白表达分别降低至75.03%、82.15%和16.59%。同时,细胞生长受到抑制,并发生细胞凋亡,凋亡率分别为32.51%、26.81%和47.19%。结论siRNA可以有效抑制HS-SY-II细胞系中融合基因SYT-SSX的表达,从而抑制细胞生长,并导致细胞凋亡。Objective To investigate the effect of specific small interfering RNA （siRNA） targeting against SYT-SSX fusion gene on the biological traits of synovial sarcoma cell line HS-SY-Ⅱ. Methods Three siRNAs against the SYT-SSX gene were designed and transfected into HS-SY-Ⅱ cells respectively. SYT-SSX gene expression was measured using RT-PCR, and fusion protein expression was analysed using Western blot. Cell proliferation was measured with MTT assay. The apoptotic cells were stained with Annexin-FITC/PI and analysed by flow cytometry. Results SYT-SSX mRNA was completely blocked by one of siRNA, and 62.7% and 29.2% attenuating of fusion gene expression were observed after transfecting with the other two siRNA respectively. The corresponding decrease of SYT-SSX protein expression was confirmed by Western blot analysis. As aconsequence, cell growth inhibition was observed, 10.51%, 7.81% and 35.72% of cells apoptosis were observed after transfecting with three siRNA respectively. Conclusions These results demonstrated that siRNA against SYT-SSX could effectively silence or downregulate fusion gene expression in HS-SY-Ⅱ cell line, and inhibit cell growth and induce cell apoptosis.

关 键 词：滑膜肉瘤 siRNA SYT-SSX 融合基因 生物治疗 

分 类 号：R738[医药卫生—肿瘤]