Targeting tumor gene by shRNA-expressing Salmonella-mediated RNAi
Targeting tumor gene by shRNA-expressing Salmonella-mediated RNAi
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DOI：
10.1038/gt.2010.112
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发表时间：
2011-01-01
期刊：
影响因子：
5.1
通讯作者：
Inal, C.
中科院分区：
文献类型：
--
作者：
Guo, H.;Zhang, J.;Inal, C.
RNA干扰（RNA interference，RNAi）是基因治疗的重要研究手段，具有巨大的应用潜力。然而，体内RNAi的靶向诱导遇到了重大挑战。在这项研究中，一个新的pSLS质粒能够表达短发夹RNA（shRNA）转化减毒沙门氏菌肠道血清型鼠伤寒7207（SL）。用转化的S.含有pSLS的肠毒素（SL-pSLS-CAT）证实靶向CTNNB 1基因的shRNA的表达在培养的SW 480细胞中诱导CTNNB 1表达的有效和特异性沉默。与对照感染细胞相比，SW 480细胞中CTNNB 1敲低与显著降低的增殖和细胞死亡相关。此外，SL-pSLS-CAT介导的CTNNB 1敲低显著降低了SW 480异种移植小鼠中的肿瘤生长。这些肿瘤表现出CTNNB 1以及c-Myc和细胞周期蛋白D1水平降低。最后，SL-pSLS-CAT处理还导致这些基因在APC（Min）小鼠的息肉、粘膜组织和小肠中的表达水平降低。总之，这些数据表明，减毒的shRNA表达沙门氏菌可能是一个强大的新工具，在体外基因沉默，功能基因组学，和基于RNAi的抗癌或人类免疫缺陷病毒治疗的发展。Gene Therapy（2011）18，95-105; doi：10.1038/gt.2010.112; 2010年9月2日在线发表
RNA interference (RNAi) has been established as an important research tool that carries great potential for gene therapy. However, targeted induction of RNAi in vivo has met with significant challenges. In this study, a novel pSLS plasmid capable of expressing short hairpin RNAs (shRNAs) was transformed into attenuated Salmonella enterica serovar typhimurium strain 7207 (SL). In vitro infection studies with the transformed S. enterica containing pSLS (SL-pSLS-CAT) demonstrated that expression of shRNA targeting the CTNNB1 gene induced potent and specific silencing of CTNNB1 expression in cultured SW480 cells. CTNNB1 knockdown in SW480 cells was associated with markedly reduced proliferation and cell death compared with that of control infected cells. In addition, SL-pSLS-CAT-mediated CTNNB1 knockdown markedly reduced tumor growth in SW480 xenograft mice. These tumors exhibited reduced levels of CTNNB1, as well as c-Myc and cyclin D1. Finally, SL-pSLS-CAT treatment also resulted in reduced expression levels of these genes in polyps, mucosal tissues and in small intestines of APC(Min) mice. Taken together, these data suggest that attenuated shRNA-expressing Salmonella may be a powerful new tool for in vitro gene silencing, functional genomics, and the development of RNAi-based anticancer or human immunodeficiency virus therapeutics. Gene Therapy (2011) 18, 95-105; doi: 10.1038/gt.2010.112; published online 2 September 2010