INHIBITION OF PAPILLOMAVIRUSES IN ORAL EPITHELIUM

人乳头瘤病毒（HPV）被认为是重要的病因 口腔癌 它们存在于许多肿瘤中， 口腔上皮细胞恶性表型。 在目前的赠款中， 在此期间，我们已经开发了切割HPV-18的RNA转录物的核酶， 并抑制宫颈癌细胞系的恶性表型 表达HPV-18的病毒 在下一个赠款期间，我们将扩大这一点， 努力实现三个目标。 具体目标1是拓宽频谱 抗HPV核酶 这将通过制造新的核酶来实现， 抑制60%以上口腔癌中发现的HPV。 病毒载体 将在一个单一的细胞中表达所有三种核酶， 构建体 该多核酶载体的功能将进一步优化 从而表达最强的抗HPV活性。 具体目标2将 使用核酶研究HPV在口腔癌生长中的作用 细胞 一大块口腔癌细胞和其他细胞将被 考察 我们将检验HPV表达病毒E6的假设， 和E7蛋白，其反过来又阻断细胞p53的活性， RB蛋白，从而防止癌细胞的程序性死亡。 具体目标3将是发现抗HPV核酶是否作为基因具有活性 口腔癌的治疗药物。 口腔癌将从 患者并移植到裸鼠体内。 在那里，他们将得到 表达抗HPV核酶的病毒载体。 抗癌效果 将用于预测这种形式的基因治疗对 口腔癌患者。 这个项目将回答基本的问题 关于口腔癌的病因学，并将有助于 开发新的治疗方法。

Human papillomaviruses (HPVs) are believed to be important in the etiology of oral cancer. They are present in many of the tumors and can transform oral epithelial cells to a malignant phenotype. In the current grant period we have developed ribozymes which cut the RNA transcript of HPV-18, and which inhibit the malignant phenotype of a cervical cancer cell line that expresses HPV-18. In the next grant period we will expand upon this work to achieve three aims. Specific Aim 1 will be to broaden the spectrum of anti-HPV ribozymes. This will be done by making new ribozymes that inhibit HPV that are found in over 60% of oral cancers. A virus vector will be made which expresses all three of the ribozymes in a single construct. The functions of this multi-ribozyme vector will be opitmized so as to express the strongest anti-HPV activity. Specific Aim 2 will be to use the ribozymes to study the role of HPVs in the growth of oral cancer cells. A large panel of oral cancer cells and other cells will be examined. We will test the hypothesis that the HPVs express the viral E6 and E7 proteins, which in turn block the activity of the cellular p53 and RB proteins, thus preventing programmed death of the cancer cells. Specific Aim 3 will be to find if the anti-HPV ribozymes are active as gene therapy agents against oral cancer. Oral cancers will be removed from patients and transplanted to nude mice. There they will be treated with the virus vector that express anti-HPV ribozymes. The anti-cancer effects will be used to predict the effects of this form of gene therapy on patients with oral cancer. This project will answer basic questions regarding the etiology of oral cancer and will contribute to the development of new treatments.