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Published on: March 11, 2014
[Inhibition of HPV16 E6 oncogene in cervical cancer by RNA interference]
Xiao-yu Niu1, Zhi-lan Peng, Wei-qiang Duan
1Department of Obstetrics and Gynecology, West China Second Hospital, Sichuan University, Chengdu 610041, China.
Objective:
The efficiency of HPV16 E6 gene silenced by RNA interference in vitro and in vivo was assessed.
Methods:
The specific siRNA of HPV16 E6 was designed and transfected into CaSki cells by liposome. Cell apoptotic rates and the changes in HPV16 E6 mRNA and protein before and after transfection were measured. Cervical cancer nude mice models were set up, siRNA was injected directly into subcutaneous tumor. The function of siRNA was evaluated by the changes in tumor volume, HPV16 E6 protein expression and apoptosis of tumor cells.
Results:
In vitro research, the cell apoptotic rates were 7.7%, 11.8%, 37.4% and 12.6% respectively at 24 h, 48 h, 5th day and 9th day after transfection. The HPV16 E6 mRNA was reduced by 77%, 83%, 59% and 41% at 24 h, 48 h, 5th day and 9th day after transfection. The inhibition rates of E6 protein measured by Flow cytometry were 79.7%, 80.4%, 71.3% and 57.4% at 24 h, 48 h, 5th day and 9th day after transfection, which were confirmed by the results of Western blot. In vivo research, E6 siRNA administration groups had great power in inhibiting tumor growth, restraining E6 protein expression, increasing tumor necrosis and apoptosis. The result of repeated injections of siRNA was better than that of single injection.
Conclusion:
RNA interference with HPV16 E6 is specific and highly efficient in vitro and in vivo.
Insights
RNA interference targeting the HPV16 E6 gene effectively silenced gene expression both in vitro and in vivo. This approach demonstrated significant potential for controlling cervical cancer progression by inducing apoptosis and inhibiting tumor growth.
Area of Science:
- Molecular Biology
- Oncology
- Gene Silencing Technologies
Context:
- Cervical cancer is a significant global health concern, often driven by persistent infection with high-risk human papillomavirus (HPV) types.
- The HPV16 E6 oncoprotein plays a crucial role in viral oncogenesis by disrupting tumor suppressor proteins.
- Developing targeted therapies to inhibit E6 function is a key strategy in cervical cancer treatment.
Purpose:
- To evaluate the efficacy of RNA interference (RNAi) targeting the HPV16 E6 gene.
- To assess the in vitro and in vivo efficiency of HPV16 E6 gene silencing using small interfering RNA (siRNA).
Summary:
- Specific siRNA targeting HPV16 E6 was designed and delivered to CaSki cells and cervical cancer xenografts.
- In vitro studies showed significant reduction in HPV16 E6 mRNA and protein levels, leading to increased cell apoptosis.
- In vivo experiments demonstrated that E6 siRNA administration inhibited tumor growth, reduced E6 protein expression, and promoted tumor necrosis and apoptosis, with repeated injections yielding superior results.
Impact:
- This study confirms that RNA interference is a specific and highly efficient method for silencing HPV16 E6.
- The findings suggest a promising therapeutic strategy for cervical cancer by targeting the HPV16 E6 oncogene.
- Successful in vivo gene silencing highlights the potential of siRNA-based therapies in clinical oncology.
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