RNAi-Mediated Knockdown of Cottontail Rabbit Papillomavirus Oncogenes Using Low-Toxicity Lipopolyplexes as a Paradigm to Treat Papillomavirus-Associated Cancers
The cottontail rabbit papillomavirus (CRPV)-associated VX2 carcinoma of the New Zealand White rabbit serves as a model system for human papillomavirus (HPV)-associated head and neck squamous cell carcinomas (HNSCCs). The aim of this study was to evaluate the tumor-inhibiting effect of RNAi-mediated...
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MDPI AG
2023-09-01
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| Online Access: | https://www.mdpi.com/1999-4923/15/10/2379 |
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| author | Uzma Ali Michael Bette Ghazala Ambreen Shashank R. Pinnapireddy Imran Tariq André Marquardt Boris A. Stuck Udo Bakowsky Robert Mandic |
| author_facet | Uzma Ali Michael Bette Ghazala Ambreen Shashank R. Pinnapireddy Imran Tariq André Marquardt Boris A. Stuck Udo Bakowsky Robert Mandic |
| author_sort | Uzma Ali |
| collection | DOAJ |
| description | The cottontail rabbit papillomavirus (CRPV)-associated VX2 carcinoma of the New Zealand White rabbit serves as a model system for human papillomavirus (HPV)-associated head and neck squamous cell carcinomas (HNSCCs). The aim of this study was to evaluate the tumor-inhibiting effect of RNAi-mediated knockdown of the CRPV oncogenes, E6 and E7, using siRNA-loaded lipopolyplexes (LPPs). VX2-carcinoma-derived cells were cultured for up to 150 passages. In addition, CRPV E6 and E7 oncogenes were transiently expressed in COS-7 cells. Efficiency and safety of LPPs were evaluated in both VX2 cells and the COS-7 cell line. Both of these in vitro CRPV systems were validated and characterized by fluorescence microscopy, Western blot, and RT-qPCR. Efficient knockdown of CRPV E6 and E7 was achieved in VX2 cells and COS-7 cells pretransfected with CRPV E6 and E7 expression vectors. Knockdown of CRPV oncogenes in VX2 cells resulted in reduced viability, migration, and proliferation and led to a G0/G1 block in the cell cycle. CRPV E6 and E7 siRNA-loaded LPPs could represent promising therapeutic agents serving as a paradigm for the treatment of papillomavirus-positive cancers and could be of value for the treatment of CRPV-associated diseases in the rabbit such as papillomas and cancers of the skin. |
| first_indexed | 2024-03-10T20:58:47Z |
| format | Article |
| id | doaj.art-8088d61dc9fd45a4b09a65c197dcb63a |
| institution | Directory Open Access Journal |
| issn | 1999-4923 |
| language | English |
| last_indexed | 2024-03-10T20:58:47Z |
| publishDate | 2023-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pharmaceutics |
| spelling | doaj.art-8088d61dc9fd45a4b09a65c197dcb63a2023-11-19T17:43:41ZengMDPI AGPharmaceutics1999-49232023-09-011510237910.3390/pharmaceutics15102379RNAi-Mediated Knockdown of Cottontail Rabbit Papillomavirus Oncogenes Using Low-Toxicity Lipopolyplexes as a Paradigm to Treat Papillomavirus-Associated CancersUzma Ali0Michael Bette1Ghazala Ambreen2Shashank R. Pinnapireddy3Imran Tariq4André Marquardt5Boris A. Stuck6Udo Bakowsky7Robert Mandic8Department of Pharmaceutics and Biopharmaceutics, Philipps-Universität Marburg, 35037 Marburg, GermanyInstitute of Anatomy and Cell Biology, Philipps-Universität Marburg, 35037 Marburg, GermanyDepartment of Pharmaceutics and Biopharmaceutics, Philipps-Universität Marburg, 35037 Marburg, GermanyDepartment of Pharmaceutics and Biopharmaceutics, Philipps-Universität Marburg, 35037 Marburg, GermanyDepartment of Pharmaceutics and Biopharmaceutics, Philipps-Universität Marburg, 35037 Marburg, GermanyDepartment of Pathology, Klinikum Stuttgart, 70174 Stuttgart, GermanyDepartment of Otorhinolaryngology, Head and Neck Surgery, University Hospital Marburg, Philipps-Universität Marburg, 35043 Marburg, GermanyDepartment of Pharmaceutics and Biopharmaceutics, Philipps-Universität Marburg, 35037 Marburg, GermanyDepartment of Otorhinolaryngology, Head and Neck Surgery, University Hospital Marburg, Philipps-Universität Marburg, 35043 Marburg, GermanyThe cottontail rabbit papillomavirus (CRPV)-associated VX2 carcinoma of the New Zealand White rabbit serves as a model system for human papillomavirus (HPV)-associated head and neck squamous cell carcinomas (HNSCCs). The aim of this study was to evaluate the tumor-inhibiting effect of RNAi-mediated knockdown of the CRPV oncogenes, E6 and E7, using siRNA-loaded lipopolyplexes (LPPs). VX2-carcinoma-derived cells were cultured for up to 150 passages. In addition, CRPV E6 and E7 oncogenes were transiently expressed in COS-7 cells. Efficiency and safety of LPPs were evaluated in both VX2 cells and the COS-7 cell line. Both of these in vitro CRPV systems were validated and characterized by fluorescence microscopy, Western blot, and RT-qPCR. Efficient knockdown of CRPV E6 and E7 was achieved in VX2 cells and COS-7 cells pretransfected with CRPV E6 and E7 expression vectors. Knockdown of CRPV oncogenes in VX2 cells resulted in reduced viability, migration, and proliferation and led to a G0/G1 block in the cell cycle. CRPV E6 and E7 siRNA-loaded LPPs could represent promising therapeutic agents serving as a paradigm for the treatment of papillomavirus-positive cancers and could be of value for the treatment of CRPV-associated diseases in the rabbit such as papillomas and cancers of the skin.https://www.mdpi.com/1999-4923/15/10/2379head and neck cancerHNSCCVX2 carcinomapapillomavirusCRPVE6 |
| spellingShingle | Uzma Ali Michael Bette Ghazala Ambreen Shashank R. Pinnapireddy Imran Tariq André Marquardt Boris A. Stuck Udo Bakowsky Robert Mandic RNAi-Mediated Knockdown of Cottontail Rabbit Papillomavirus Oncogenes Using Low-Toxicity Lipopolyplexes as a Paradigm to Treat Papillomavirus-Associated Cancers Pharmaceutics head and neck cancer HNSCC VX2 carcinoma papillomavirus CRPV E6 |
| title | RNAi-Mediated Knockdown of Cottontail Rabbit Papillomavirus Oncogenes Using Low-Toxicity Lipopolyplexes as a Paradigm to Treat Papillomavirus-Associated Cancers |
| title_full | RNAi-Mediated Knockdown of Cottontail Rabbit Papillomavirus Oncogenes Using Low-Toxicity Lipopolyplexes as a Paradigm to Treat Papillomavirus-Associated Cancers |
| title_fullStr | RNAi-Mediated Knockdown of Cottontail Rabbit Papillomavirus Oncogenes Using Low-Toxicity Lipopolyplexes as a Paradigm to Treat Papillomavirus-Associated Cancers |
| title_full_unstemmed | RNAi-Mediated Knockdown of Cottontail Rabbit Papillomavirus Oncogenes Using Low-Toxicity Lipopolyplexes as a Paradigm to Treat Papillomavirus-Associated Cancers |
| title_short | RNAi-Mediated Knockdown of Cottontail Rabbit Papillomavirus Oncogenes Using Low-Toxicity Lipopolyplexes as a Paradigm to Treat Papillomavirus-Associated Cancers |
| title_sort | rnai mediated knockdown of cottontail rabbit papillomavirus oncogenes using low toxicity lipopolyplexes as a paradigm to treat papillomavirus associated cancers |
| topic | head and neck cancer HNSCC VX2 carcinoma papillomavirus CRPV E6 |
| url | https://www.mdpi.com/1999-4923/15/10/2379 |
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