New highly potent and specific E6 and E7 siRNAs for treatment of HPV16 positive cervical cancer
New highly potent and specific E6 and E7 siRNAs for treatment of HPV16 positive cervical cancer
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DOI：
10.1038/sj.cgt.7701118
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发表时间：
2008-03-01
影响因子：
6.4
通讯作者：
Yoshinouchi, M.
中科院分区：
文献类型：
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作者：
Yamato, K.;Yamada, T.;Yoshinouchi, M.
高危型人乳头瘤病毒（HPV）的持续感染是宫颈癌的必要原因，其中HPV16最为普遍，占报告病例的50%以上。该病毒编码E6和E7癌蛋白，其表达对于维持恶性表型至关重要。为了选择适用于HPV16+宫颈癌患者RNAi治疗的有效sirna，使用siDirect计算机软件设计E6和E7 sirna，然后选择10个与所有HPV16变体兼容的sirna，然后使用HPV16+和HPV16-细胞广泛检测RNAi活性和特异性。选择对E6和E7表达RNAi活性最高的3种sirna，对HPV16+癌细胞低至1 nM的生长具有特异性和有效的抑制作用。生长抑制伴随着p53和p21（WAF1/CIP1）的积累，以及细胞衰老特征的形态学和细胞化学变化。其中一种sirna的抗肿瘤活性被证实，当局部注射与间胶原复合物时，NOD/SCID小鼠HPV16+细胞的肿瘤生长被延缓。我们的研究结果表明，这些E6和E7 sirna是治疗病毒相关癌症的有希望的治疗药物。
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 nM 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.