PATHOBIOLOGY OF CERVICAL INTRAEPITHELIAL NEOPLASIA

宫颈上皮内瘤变的病理学

• 批准号: 3191457
• 负责人: Christopher Paul Crum
• 金额: $ 13.3万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-06-01 至 1993-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3191457
• 关键词: Papillomavirus; cervix neoplasms; gene expression; genetic manipulation; genetic mapping; genetic recombination; genetic transcription; human tissue; laboratory rabbit; molecular cloning; neoplasm /cancer invasiveness; nucleic acid hybridization; open reading frames; preneoplastic state; squamous cell carcinoma; transforming virus; viral carcinogenesis; virus genetics

Genital papillomaviruses are associated with a spectrum of changes in the squamous epithelium, including condylomata, precancers (intraepithelial neoplasms) and invasive carcinomas. Specific HPV types, such as type 16, are associated with genital precancers, which, in contrast to condyloma, have a high risk of progressing to invasion. In this project, we will investigate HPV 16 gene expression in genital precancers to determine if HPV 16 codes unique transcripts or translation products in precancers which may account for differences in their biology vis a vis condylomata. To accomplish this, we will analyze transcription of HPV 16 open reading frames (ORF's) in precancers by RNA-RNA in-situ hybridization and cDNA cloning. The former will make it possible to localize transcription of individual ORF's in the epithelium and the latter will more precisely identify the structure of the transcripts produced. Specific early ORF's of HPV 16 which are known to be expressed in precancers, including those which are analogous to transforming genes in bovine papillomavirus (E4, E5, E6) will be cloned into Path vectors and the Tryp-fusion proteins produced will be used to generate polycolonal antibodies in rabbits. These will be reacted with tissue samples by western blot analysis and immunohistochemistry. These studies will aid in defining precisely the HPV gene products produced in the early stages of genital squamous neoplasia and will provide reagents which will be useful for further analysis of their function. Finally, to elucidate the host response to HPV infection, we will attempt to transform established cell lines (NIH 3T3, CREF) with high molecular weight DNA from precancers. Transforming sequences will be characterized in an effort to establish a profile of genes which are necessary to the process of HPV mediated neoplastic transformation in genital squamous epithelium.

生殖器乳头瘤病毒与一系列相关 鳞状上皮的变化，包括尖锐湿疣， 癌前病变（上皮内肿瘤）和浸润性癌。 特定 HPV 类型（例如 16 型）与生殖器相关 与尖锐湿疣相比，癌前病变的风险很高 进展到入侵。 在这个项目中，我们将研究 HPV 生殖器癌前病变中 16 基因的表达以确定是否 HPV 16 编码癌前病变中独特的转录本或翻译产物 这可能解释了它们在生物学上的差异 尖锐湿疣。 为了实现这一点，我们将分析 通过 RNA-RNA 检测癌前病变中的 HPV 16 开放阅读框 (ORF) 原位杂交和cDNA克隆。 前者会成功 可以将单个 ORF 的转录定位在 上皮细胞和后者将更准确地识别 产生的转录本的结构。 特定的早期ORF 已知在癌前病变中表达的 HPV 16，包括 那些类似于牛的转化基因 乳头瘤病毒（E4、E5、E6）将被克隆到 Path 载体中并 产生的胰蛋白酶融合蛋白将用于生成 兔多克隆抗体。 这些将发生反应 通过蛋白质印迹分析组织样本和 免疫组织化学。 这些研究将有助于定义 正是HPV病毒早期产生的基因产物 生殖器鳞状肿瘤并将提供试剂 对于进一步分析它们的功能很有用。 最后，要说明的是 宿主对HPV感染的反应，我们将尝试改变 已建立的高分子量细胞系（NIH 3T3、CREF） 来自癌前病变的 DNA。 变换序列将是 其特点是努力建立基因谱， 是 HPV 介导的肿瘤过程所必需的 生殖器鳞状上皮的转化。