EBV EXPRESSION IN NASOPHARYNGEAL CARCINOMA

鼻咽癌中 EBV 的表达

• 批准号: 3170879
• 负责人: NANCY JOAN RAAB-TRAUB
• 金额: $ 14.62万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-06-01 至 1991-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3170879
• 关键词: Epstein Barr virus; bacteriophage lambda; epithelium; gene expression; genetic library; genetic manipulation; genetic mapping; genetic promoter element; genetic transcription; hyperplasia; messenger RNA; molecular cloning; nasopharyngeal neoplasms; neoplasm /cancer genetics; nucleic acid hybridization; nucleic acid sequence; oncogenic virus; preneoplastic state; tissue /cell culture; viral carcinogenesis; virus DNA; virus RNA; virus genetics

Nasopharyngeal carcinoma (NPC) is an epithelial malignancy which occurs at high incidence in endemic regions in Southern China, Northern Africa, and among Alaskan Eskimoes but also occurs worldwide in all populations. We have identified the viral sequences which encode RNA in tumor tissue and have identified specific viral transcripts from three regions of the EBV genome which are transcribed in NPC. We will determine when these transcripts are synthesized in lymphoid cell lines in vitro and in the NPC epithelial fusion cell line. NPC-KT. In order to further define which viral functions are expressed in malignant epithelial tissues we will use single-stranded RNA probes representing specific latent functions generated from vectors containing the SP6 and T7 bacteriophage promoters in Northern analyses or in nuclease protection assays. RNA preparations from NPC biopsy specimens an NPC which can be passaged in nude mice, C15, and the NPC cell line, NPC-KT will be compared to RNA from lymphoid cell lines. To facilitate a detailed analysis of viral transcription from the entire genome, we have constructed a cDNA library to NPC RNA in the vector, lambda gt11, by priming cDNA synthesis with oligo dT. Twelve viral specific clones have been identified. The viral cDNA structures will determined by restriction enzyme and sequence analysis. Additional libraries will be repeated from RNA from the same specimen and from additional NPC samples. In some instances cDNA will be primed with oligonucleotides for viraal genes. We will identify hyperplasias or other oropharyngeal malignancies associated with EBV by screening for EBV DNA. The structure of the viral termini will be analyzed to determine if the infected tissue produces linear virion DNA or contains only the episomal form. Monoclonal proliferations will be identified by this assay. Viral transcripts will be analyzed in the EBV-positive hyperplasias through the use of Northern blots, nuclease protection assays and cDNA cloning.

鼻咽癌（NPC）是一种上皮性恶性肿瘤 该病在南方流行地区发病率较高 中国、北非和阿拉斯加爱斯基摩人 发生在全世界所有人群中。 我们已经确定了病毒 编码肿瘤组织中 RNA 的序列已被鉴定 来自 EBV 基因组三个区域的特定病毒转录本 是在 NPC 中转录的。 我们将确定何时这些 转录物在体外和体内的淋巴细胞系中合成 鼻咽癌上皮融合细胞系。 NPC-KT。 为了进一步明确表达哪些病毒功能 恶性上皮组织我们将使用单链RNA 代表特定潜在功能的探针生成 含有 SP6 和 T7 噬菌体启动子的载体 Northern 分析或核酸酶保护测定。 核糖核酸 鼻咽癌活检标本的制备物 在裸鼠、C15 和 NPC 细胞系 NPC-KT 中传代 与来自淋巴细胞系的 RNA 进行比较。 为了便于对病毒转录进行详细分析 整个基因组，我们构建了 NPC RNA 的 cDNA 文库 在载体 lambda gt11 中，通过用寡核苷酸引发 cDNA 合成 dT。 已鉴定出十二个病毒特异性克隆。 病毒式传播 cDNA 结构将通过限制性内切酶确定 序列分析。 其他库将重复从 来自同一样本和其他 NPC 样本的 RNA。 在某些情况下，cDNA 将用寡核苷酸引发 病毒基因。 我们将识别增生或其他口咽恶性肿瘤 通过筛查 EBV DNA 与 EBV 相关。 的结构 将分析病毒末端以确定是否被感染 组织产生线性病毒粒子DNA或仅包含游离病毒粒子 形式。 单克隆增殖将通过该测定来鉴定。 将在 EBV 阳性增生中分析病毒转录本 通过使用 Northern 印迹、核酸酶保护测定和 cDNA克隆。