壳聚糖-三聚磷酸钠纳米颗粒介导的shRNA递送对TGFB1基因沉默最佳位点的选择

Selection of optimal sites for TGFB1 gene silencing by chitosan-TPP nanoparticle-mediated delivery of shRNA.

作者信息

Wang So-Ly, Yao Hui-Hua, Guo Ling-Ling, Dong Liang, Li Shi-Gang, Gu Yong-Ping, Qin Zheng-Hong

机构信息

Department of Pathology, Soochow University School of Medicine, Suzhou, China.

出版信息

Cancer Genet Cytogenet. 2009 Apr 1;190(1):8-14. doi: 10.1016/j.cancergencyto.2008.10.013.

DOI:10.1016/j.cancergencyto.2008.10.013

PMID:19264227

Abstract

Most human tumors produce high levels of TGF-beta1, whose autocrine and paracrine actions promote tumor cell invasiveness and metastasis. Currently, many experimental therapies that target TGFB1 have utilized antisense DNA or RNA interference (RNAi). Despite the great potential of RNAi, the selection of effective target sites and proper delivery systems for short hairpin RNA (shRNA) remains a significant issue. Here, we used chitosan nanoparticle-mediated delivery of a shRNA-expressing vector to inhibit TGFB1 expression in the human rhabdomyosarcoma cell line RD. Knockdown of TGFB1 by shRNA resulted in a decrease in RD cell growth in vitro and tumorigenicity in nude mice. The efficiency of TGFB1 gene silencing varied with the selection of targeting sites. These data suggest that chitosan nanoparticle-mediated delivery of an shRNA produces efficient TGFB1 knockdown in rhabdomyosarcoma cells and may be a method of choice for shRNA delivery for gene therapy.

摘要

大多数人类肿瘤会产生高水平的转化生长因子β1（TGF-β1），其自分泌和旁分泌作用会促进肿瘤细胞的侵袭和转移。目前，许多针对TGFB1的实验性疗法都采用了反义DNA或RNA干扰（RNAi）技术。尽管RNAi具有巨大潜力，但为短发夹RNA（shRNA）选择有效的靶位点和合适的递送系统仍然是一个重大问题。在此，我们使用壳聚糖纳米颗粒介导递送表达shRNA的载体，以抑制人横纹肌肉瘤细胞系RD中的TGFB1表达。shRNA敲低TGFB1导致RD细胞在体外的生长以及在裸鼠中的致瘤性降低。TGFB1基因沉默的效率因靶位点的选择而异。这些数据表明，壳聚糖纳米颗粒介导递送shRNA可在横纹肌肉瘤细胞中有效敲低TGFB1，并且可能是用于基因治疗的shRNA递送的一种选择方法。