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Abstract
Conclusion: Our results indicate that siRNA E6 and/or E7 may have potential as a gene-specific therapy for human papillomavirus (HPV) type 16 (HPV16)-related squamous cell carcinoma of the head and neck (HNSCC). Objectives: To evaluate the effectiveness of siRNA targeting E6 and/or E7 on the in vitro and in vivo growth suppression of HPV16-related HNSCC. Methods: HPV16-related HNSCC (UM-SCC47) cell lines were used for the present study. Expression of HPV viral oncogenes E6 and/or E7 and their cellular targets, p53 and pRb, was evaluated by real-time PCR, Western blotting, and immunofluorescence staining. To study the effect of siRNA on tumor growth in vivo, we developed animal models. Representative tumors harvested from each group were processed for apoptosis analyses (TUNEL assay) and immunofluorescence staining for p53 and pRb. Results: E6 and E7 oncogenes of HPV16 were down-regulated by E6 and/or E7 targeting siRNAs, respectively. The expression of p53 and pRb proteins in both the E6 siRNA group and E7 siRNA group was up-regulated compared with those of control groups. The cellular proliferation and apoptosis indexes of E6 and/or E7 siRNA groups were higher than those of controls. In vivo studies showed significant inhibitory effect of E6 and/or E7 siRNA compared with those of control groups, which was consistent with in vitro studies.
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Acknowledgments
We express our gratitude to Dr Thomas E. Carey and Dr Carol R. Bradford, University of Michigan, for kindly providing the UM-SCC47 cell lines and H. Matsunaga for assisting with immunohistochemical analyses. This research was supported by Grants-in-aid for Scientific Research (C) from Japan Society for the Promotion of Science; project no. 20592016 (N.O.), and project numbers 20251283 and 21390462 (K.N.). This study was presented at the 2011 Annual Meeting of the American Academy of Otolaryngology-Head and Neck Surgery Foundation, September 11–14, 2011 in San Francisco, CA.
Declaration of interest: The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper.