Retrovirus-mediated delivery of HPV16 E7 antisense RNA inhibited tumorigenicity of CaSki cells.

In cervical cancer, high-risk human papillomavirus (HPV) genes are expressed solely in cancerous cells and have been proposed to be the most important etiological factors for cervical cancer, thus making them suitable targets for gene therapy. In this study, we aim to inactivate the HPV16 E7 in CaSki cells and test the possibility of reducing the tumorigenicity of these cells. The full-length HPV16 E7 cDNA was cloned in the pBabe-puro or pWZL-Hygro retrovirus vector in reverse orientation and was stably transfected into CaSki cells by replication-defective retrovirus infection giving rise to CaSki-E7AS and CaSki-E7AS2X cells. Immunoprecipitation/Western analysis and real-time RT-PCR were performed to document the levels of HPV16 E7 gene product. Flow cytometry was performed to study changes in the cell cycle in response to reduced E7 protein. The expression of bcl-2, RB, and E2F-1 was studied using Western blot analysis. Tumorigenicity of CaSki, CaSki-E7AS, and CaSki-E7AS2X cells was assayed with subepidermal tumor growth in nude mice. We have documented that the delivery of the antisense gene construct resulted in the reduction of HPV16 E7 protein expression and cell proliferation in CaSki cells. Furthermore, we demonstrated that these changes were accompanied by cell cycle arrest, up-regulation of RB, and down-regulation of E2F-1 and bcl-2 proteins. More importantly, dose-dependent transduction of the antisense HPV16E7 construct was able to inhibit and/or retard the tumorigenicity of CaSki cells in vivo. Down-regulation of HPV16 E7 with antisense RNA is beneficial in reducing the tumorigenicity of CaSki cells and can potentially be useful for HPV-associated malignancy gene therapy.

Choo CK ，Ling MT ，Suen CK ，Chan KW ，Kwong YL ... -  《GYNECOLOGIC ONCOLOGY》

Effect of BPV1 E2-mediated inhibition of E6/E7 expression in HPV16-positive cervical carcinoma cells.

Moon MS ，Lee CJ ，Um SJ ，Park JS ，Yang JM ，Hwang ES ... -  《GYNECOLOGIC ONCOLOGY》

Reversal of the malignant phenotype of cervical cancer CaSki cells through adeno-associated virus-mediated delivery of HPV16 E7 antisense RNA.

Wu S ，Meng L ，Wang S ，Wang W ，Xi L ，Tian X ，Chen G ，Wu Y ，Zhou J ，Xu G ，Lu Y ，Ma D ... -  《CLINICAL CANCER RESEARCH》

Suppression of tumorigenesis by transcription units expressing the antisense E6 and E7 messenger RNA (mRNA) for the transforming proteins of the human papilloma virus and the sense mRNA for the retinoblastoma gene in cervical carcinoma cells.

Hu G ，Liu W ，Hanania EG ，Fu S ，Wang T ，Deisseroth AB ... -  《CANCER GENE THERAPY》

Adenovirus-mediated transfer of HPV 16 E6/E7 antisense RNA to human cervical cancer cells.

Hamada K ，Sakaue M ，Alemany R ，Zhang WW ，Horio Y ，Roth JA ，Mitchell MF ... -  《GYNECOLOGIC ONCOLOGY》