In vitro and in vivo growth suppression of human papillomavirus 16-positive cervical cancer cells by CRISPR/Cas9
In vitro and in vivo growth suppression of human papillomavirus 16-positive cervical cancer cells by CRISPR/Cas9
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DOI：
10.1016/j.bbrc.2014.07.014
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发表时间：
2014-08-08
影响因子：
3.1
通讯作者：
Li, Yan
中科院分区：
文献类型：
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作者：
Zhen, Shuai;Hua, Ling;Li, Yan
高危人乳头瘤病毒癌基因（E6和E7）的表达失控是宫颈癌发病和进展的关键事件。因此，这两种病毒致癌基因被认为是理想的治疗靶点。为了开发HPV相关癌症的基因特异性治疗方法，我们建立了靶向HPV 16启动子E6/E7的CRISPR/Cas9，并靶向E6、E7转录本，将CRISPR/Cas9转导到宫颈HPV-16阳性细胞株SiHa中。结果表明，CRISPR/Cas9靶向启动子，以及靶向E6和E7，导致p53和p21蛋白的积累，从而显著降低体外宫颈癌细胞的增殖能力。然后我们将细胞皮下接种到裸鼠体内，建立移植瘤动物模型，发现CRISPR/Cas9靶向（启动子+E6+E7）转录物的细胞孵育后，小鼠的肿瘤发生和生长明显受到抑制。我们的研究结果可能为靶向HR-HPV关键癌基因的CRISPR/Cas9作为一种新的治疗策略在宫颈癌和其他hpv相关癌症治疗中的应用提供证据。(C) 2014爱思唯尔公司版权所有。
Deregulated expression of high-risk human papillomavirus oncogenes (E6 and E7) is a pivotal event for pathogenesis and progression in cervical cancer. Both viral oncogenes are therefore regarded as ideal therapeutic targets. In the hope of developing a gene-specific therapy for HPV-related cancer, we established CRISPR/Cas9 targeting promoter of HPV 16 E6/E7 and targeting E6, E7 transcript, transduced the CRISPR/Cas9 into cervical HPV-16-positive cell line SiHa. The results showed that CRISPR/Cas9 targeting promoter, as well as targeting E6 and E7 resulted in accumulation of p53 and p21 protein, and consequently remarkably reduced the abilities of proliferation of cervical cancer cells in vitro. Then we inoculated subcutaneously cells into nude mice to establish the transplanted tumor animal models, and found dramatically inhibited tumorigenesis and growth of mice incubated by cells with CRISPR/Cas9 targeting (promoter+E6+E7)-transcript. Our results may provide evidence for application of CRISPR/Cas9 targeting HR-HPV key oncogenes, as a new treatment strategy, in cervical and other HPV-associated cancer therapy. (C) 2014 Elsevier Inc. All rights reserved.