Zinc finger nucleases targeting the human papillomavirus E7 oncogene induce E7 disruption and a transformed phenotype in HPV16/18-positive cervical cancer cells.

Cervical cancer is mainly caused by infections of high-risk human papillomavirus (HR-HPV). Persistent expression of HR-HPV oncogenes E6 and E7 is implicated in malignant transformation. The aim was to provide proof-of-concept data to support use of zinc finger nucleases (ZFN) targeting HPV E7 to treat HPV-related cervical cancer. We designed and constructed ZFNs that could specifically recognize and cleave HPV16/18 E7 DNA. We tested the cleavage efficiency of selected ZFN16-E7-S2 and ZFN18-E7-S2 by using single-strand annealing (SSA) assay. Cell viability and colony formation assays were used to estimate the inhibition of cell growth that received treatments of ZFNs. Gene disruption of HPV E7 and downstream genes were examined by Western blotting. Cell apoptosis assay was used to test the specificity and efficiency of induction of HPV type-specific apoptosis. We also introduced xenograft formation assays to estimate the potential of inhibition of HPV-related disease. We found ZFN16-E7-S2 and ZFN18-E7-S2 disrupted HPV E7 oncogenes in HPV16/18-positive cervical cancer cells. Both ZFNs effectively led to inhibition of type-specific cervical cancer cell growth, and specifically induced apoptosis of corresponding HPV16- and HPV18-positive cervical cancer cell lines. ZFN16-E7-S2 and ZFN18-E7-S2 also repressed xenograft formation in vivo. ZFNs targeting HPV16/18 E7 could effectively induce disruption of E7 oncogenes and lead to type-specific and efficient growth inhibition and apoptosis of HPV-positive cells. ZFNs targeting HPV16/18 E7 oncogenes could be used as novel therapeutic agents for the treatment of HPV-related cervical cancer.

Ding W ，Hu Z ，Zhu D ，Jiang X ，Yu L ，Wang X ，Zhang C ，Wang L ，Ji T ，Li K ，He D ，Xia X ，Liu D ，Zhou J ，Ma D ，Wang H ... -  《-》

Honeybee venom possesses anticancer and antiviral effects by differential inhibition of HPV E6 and E7 expression on cervical cancer cell line.

Kim YW ，Chaturvedi PK ，Chun SN ，Lee YG ，Ahn WS ... -  《-》

In vitro and in vivo growth inhibition of human cervical cancer cells via human papillomavirus E6/E7 mRNAs' cleavage by CRISPR/Cas13a system.

Sustained infection of high-risk human papillomavirus (HR-HPVs), especially HPV16 and HPV18, is a major cause of cervical cancer. E6 and E7 oncoproteins, encoded by the HPV genome, are critical for transformation and maintenance of malignant phenotypes of cervical cancer. Here, we used an emerging programmable clustered regularly interspaced short palindromic repeat (CRISPR)/Cas13a system to cleave HPV 16/18 E6/E7 messenger RNAs (mRNAs). The results showed that customized CRISPR/Cas13a system effectively and specifically knocked down HPV 16/18 E6/E7 mRNAs, inducing growth inhibition and apoptosis in HPV16-positive SiHa and HPV18-positive HeLa Cell lines, but not in HPV-negative C33A cell line. Simultaneously, we detected downregulation of E6/E7 oncoproteins and upregulation of tumor suppressor P53 and RB proteins. In addition, we used subcutaneous xenograft tumor growth assays to find that the weight and volume of tumors in the SiHa-16E6CR1 group knocked down by the CRISPR/Cas13a system were significantly lower than those in the SiHa-VECTOR group lacking crRNA. Our study demonstrated that targeting HPV E6/E7 mRNAs by the CRISPR/Cas13a system may be a candidate therapeutic strategy for HPV-related cervical cancer.

Chen Y ，Jiang H ，Wang T ，He D ，Tian R ，Cui Z ，Tian X ，Gao Q ，Ma X ，Yang J ，Wu J ，Tan S ，Xu H ，Tang X ，Wang Y ，Yu Z ，Han H ，Das BC ，Severinov K ，Hitzeroth II ，Debata PR ，Xu W ，Fan W ，Jin Z ，Cao C ，Yu M ，Xie W ，Huang Z ，Hu Z ，You Z ... -  《-》

RNA interference against HPV16 E7 oncogene leads to viral E6 and E7 suppression in cervical cancer cells and apoptosis via upregulation of Rb and p53.

Sima N ，Wang W ，Kong D ，Deng D ，Xu Q ，Zhou J ，Xu G ，Meng L ，Lu Y ，Wang S ，Ma D ... -  《APOPTOSIS》

TALEN-mediated targeting of HPV oncogenes ameliorates HPV-related cervical malignancy.

Hu Z ，Ding W ，Zhu D ，Yu L ，Jiang X ，Wang X ，Zhang C ，Wang L ，Ji T ，Liu D ，He D ，Xia X ，Zhu T ，Wei J ，Wu P ，Wang C ，Xi L ，Gao Q ，Chen G ，Liu R ，Li K ，Li S ，Wang S ，Zhou J ，Ma D ，Wang H ... -  《-》