New highly potent and specific E6 and E7 siRNAs for treatment of HPV16 positive cervical cancer

• Published in: Cancer gene therapy Vol. 15; no. 3; pp. 140 - 153
• Main Authors: Yamato, K, Yamada, T, Kizaki, M, Ui-Tei, K, Natori, Y, Fujino, M, Nishihara, T, Ikeda, Y, Nasu, Y, Saigo, K, Yoshinouchi, M
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01.03.2008; Nature Publishing Group
• Subjects: Animals; Antitumor activity; Atelocollagen; Biomedical and Life Sciences; Biomedicine; Care and treatment; Cell Line, Tumor; Cell Proliferation; Cellular Senescence - genetics; Cellular Senescence - physiology; Cervical cancer; Cervix; Complications and side effects; Cyclin-dependent kinase inhibitor p21; Female; Gene Expression; Gene Therapy; Genetic aspects; Genetic Therapy - methods; HeLa Cells; Human papillomavirus; Human papillomavirus 16; Human papillomavirus 16 - genetics; Human papillomavirus 16 - growth & development; Humans; Immunoblotting; Luciferases - genetics; Luciferases - metabolism; Mice; Mice, Inbred NOD; Mice, SCID; Oncogene Proteins, Viral - genetics; original-article; Papillomaviridae; Papillomavirus E7 Proteins; Papillomavirus infections; Phenotypes; Physical characteristics; Repressor Proteins - genetics; Reverse Transcriptase Polymerase Chain Reaction; Risk factors; RNA, Small Interfering - genetics; RNA, Small Interfering - physiology; RNA-mediated interference; Senescence; siRNA; Time Factors; Transfection; Tumor Burden; Uterine Cervical Neoplasms - genetics; Uterine Cervical Neoplasms - therapy; Uterine Cervical Neoplasms - virology; Xenograft Model Antitumor Assays; Japan; cervical cancer; siRNA therapy; HPV16
• ISSN: 0929-1903; 1476-5500
• DOI: 10.1038/sj.cgt.7701118

Persistent infection by high-risk types of human papillomaviruses (HPV) is a necessary cause of cervical cancer, with HPV16 the most prevalent, accounting for more than 50% of reported cases. The virus encodes the E6 and E7 oncoproteins, whose expression is essential for maintenance of the malignant phenotype. To select efficacious siRNAs applicable to RNAi therapy for patients with HPV16+ cervical cancer, E6 and E7 siRNAs were designed using siDirect computer software, after which 10 compatible with all HPV16 variants were selected, and then extensively examined for RNAi activity and specificity using HPV16+ and HPV16−cells. Three siRNAs with the highest RNAi activities toward E6 and E7 expression, as well as specific and potent growth suppression of HPV16+ cancer cells as low as 1 n M were chosen. Growth suppression was accompanied by accumulation of p53 and p21 WAF1/CIP1 , as well as morphological and cytochemical changes characteristic of cellular senescence. Antitumor activity of one of the selected siRNAs was confirmed by retarded tumor growth of HPV16+ cells in NOD/SCID mice when locally injected in a complex with atelocollagen. Our results demonstrate that these E6 and E7 siRNAs are promising therapeutic agents for treatment of virus-related cancer.