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Adenovirus-mediated transfer of HPV 16 E6/E7 antisense RNA to human cervical cancer cells
K Hamada1, M Sakaue, R Alemany
1Department of Thoracic and Cardiovascular Surgery, The University of Texas, M.D. Anderson Cancer Center, Houston 77030, USA.
Abstract:
To explore the potential of an adenoviral antisense RNA transcript for gene therapy of cervical cancer, we introduced the antisense RNA transcript of E6 and E7 genes of human papillomavirus (HPV) 16 into cervical cancer cells harboring HPV 16 via a recombinant adenoviral vector, Ad5CMV-HPV 16 AS and analyzed the effects of expression of these genes on cell growth and tumor growth. Ad5CMV-HPV 16 AS contains the cytomegalovirus-promoter, E6 and E7 genes of HPV 16 in antisense orientation, and the SV40 polyadenylation signal in a mini-gene cassette, which is inserted into the E1-deleted region of modified adenovirus 5. The entire E6/E7 region of HPV 16 was amplified by polymerase chain reaction (PCR) before cloning into the mini-gene cassette. By reverse transcriptase-PCR, HPV 16 E6/E7 antisense RNA was detected in SiHa cells infected with Ad5CMV-HPV 16 AS. The growth of the Ad5CMV-HPV 16 AS-infected cells was greatly suppressed, as evidenced by a decrease in cell count. The growth inhibitory effect of Ad5CMV-HPV 16 AS was significantly enhanced by an adenoviral p53 construct, Ad5CMV-p53. In an ex vivo study in nude mice, tumorigenicity was completely inhibited in mice injected with Ad5CMV-HPV 16 AS-infected SiHa cells. These data suggest that transfection of cervical cancer cells with HPV 16 E6/E7 antisense RNA in a form such as Ad5CMV-HPV 16 AS is a potential novel approach to the therapy of HPV 16-positive cervical cancer.
Insights
This study explores adenoviral antisense RNA therapy for cervical cancer. Introducing human papillomavirus (HPV) 16 E6/E7 antisense RNA via Ad5CMV-HPV 16 AS significantly inhibited cancer cell and tumor growth.
Area of Science:
- Oncology
- Virology
- Gene Therapy
Background:
- Cervical cancer is frequently associated with persistent infection by high-risk human papillomavirus (HPV) types, particularly HPV 16.
- The viral E6 and E7 oncoproteins play crucial roles in the oncogenesis of HPV-associated cervical cancers.
- Targeting these viral oncoproteins presents a potential strategy for cervical cancer gene therapy.
Purpose of the Study:
- To investigate the therapeutic potential of an adenoviral vector expressing antisense RNA targeting HPV 16 E6 and E7 genes.
- To evaluate the impact of HPV 16 E6/E7 antisense RNA expression on cervical cancer cell growth and tumor development.
Main Methods:
- A recombinant adenoviral vector, Ad5CMV-HPV 16 AS, was constructed to deliver HPV 16 E6/E7 antisense RNA to cervical cancer cells.
- Polymerase chain reaction (PCR) and reverse transcriptase-PCR were used to confirm gene amplification and antisense RNA expression.
- Cell proliferation assays and in vivo tumorigenicity studies in nude mice were performed to assess therapeutic efficacy.
Main Results:
- Ad5CMV-HPV 16 AS successfully delivered and expressed HPV 16 E6/E7 antisense RNA in SiHa cervical cancer cells.
- Expression of HPV 16 E6/E7 antisense RNA significantly suppressed cervical cancer cell growth.
- Co-administration with an adenoviral p53 construct (Ad5CMV-p53) enhanced the growth inhibitory effect.
- Tumorigenicity was completely inhibited in nude mice injected with Ad5CMV-HPV 16 AS-infected SiHa cells.
Conclusions:
- Adenoviral delivery of HPV 16 E6/E7 antisense RNA (Ad5CMV-HPV 16 AS) demonstrates significant anti-proliferative and anti-tumorigenic effects.
- This approach offers a promising novel therapeutic strategy for HPV 16-positive cervical cancer.
- Combination therapy with p53 may further enhance treatment outcomes.