title: In vitro and in vivo growth suppression of human papillomavirus 16-positive cervical cancer cells by CRISPR/Cas9.
abstract: 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.
authors.affiliation: Baoji Maternal and Child Health Hospital, 2 Xinjian Road East, WeiBin District, Baoji City, 721000, Shanxi Province, PR China; Xijing Hospital, Fourth Military Medical University, Xi'an, PR China; Department of Pharmacology and Toxicology, Beijing Institute of Radiation Medicine, Beijing 100850, PR China; Kyoto University, Kyoto 606-8507, Japan. Electronic address: usa_2002@163.com.
authors.forename: Shuai
authors.initials: S
authors.lastname: Zhen
authors.affiliation: Department of Pharmacology and Toxicology, Beijing Institute of Radiation Medicine, Beijing 100850, PR China.
authors.forename: Ling
authors.initials: L
authors.lastname: Hua
authors.affiliation: Kyoto University, Kyoto 606-8507, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Takahashi
authors.affiliation: Kyoto University, Kyoto 606-8507, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Narita
authors.affiliation: Department of Pharmacology and Toxicology, Beijing Institute of Radiation Medicine, Beijing 100850, PR China.
authors.forename: Yun-Hui
authors.initials: YH
authors.lastname: Liu
authors.affiliation: Baoji Hospital of Traditional Chinese Medicine, No 43, BaoFu Road, Baoji City, Shanxi Province, PR China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
mesh_terms: D052685:Alphapapillomavirus / Q000302:isolation & purification*; D001483:Base Sequence; D002455:Cell Division / Q000502:physiology*; D045744:Cell Line, Tumor; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000502:physiology*; D017931:DNA Primers; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D002583:Uterine Cervical Neoplasms / Q000473:pathology / Q000821:virology*
chemical_list: D017931:DNA Primers
keywords: BALB/C nude mouse; CRISPR/Cas9; E6; E7; HPV16; Tumor formation