TUMOR CELL DEATH INDUCED BY INHIBITION OF REPLICATION

复制抑制引起的肿瘤细胞死亡

• 批准号: 2683642
• 负责人: VICTOR V LEVENSON
• 金额: $ 10.5万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-04-05 至 2001-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2683642
• 关键词: DNA replication; HeLa cells; antineoplastics; antisense nucleic acid; aphidicolin; apoptosis; cell cycle; cell type; cellular immunity; complementary DNA; cytotoxicity; drug resistance; drug screening /evaluation; gene expression; gene induction /repression; genetic library; human genetic material tag; neoplastic cell; nucleic acid sequence; protein biosynthesis; transfection /expression vector

DESCRIPTION: Many cytotoxic drugs used in cancer chemotherapy inhibit DNA replication and induce programmed cell death (apoptosis) in tumor cells. It is unknown to what extent the inhibition of DNA replication per se is responsible for the induction of cell death by anticancer drugs. Cell death in cycling cells can be induced by treatment with known inhibitors of DNA replication, such as aphidicolin, a mycotoxin which inhibits elongation of replication by DNA polymerases alpha and delta, or mimosine, a plant amino acid which prevents initiation of replication in mammalian cells. The main goal of this application is to identify genes and genetic pathways that are involved in the induction of cell death by inhibitors of DNA replication. These genes will be identified through expression selection of genetic suppressor elements (GSE), short cDNA fragments that inhibit gene expression by encoding dominantly acting peptides or inhibitory antisense RNA. A normalized library of GSEs in a retroviral vector will be transduced into HeLa cells, and GSEs which protect cells from aphidicolin- or mimosine-induced cell death will be isolated. These GSEs will be characterized for their efficacy, effects on cell cycle progression, ability to protect cells against different DNA replication inhibitors and cell-type specificity. Full-length cDNAs for human genes corresponding to the selected GSEs will be isolated, and their structure and expression patterns in the cell cycle will be characterized. The isolated GSEs will also be tested for their effects on cellular response to replication-inhibiting chemotherapeutic drugs, to determine the role of replication-based mechanisms of cytotoxicity in their antitumor effect.

描述：癌症化疗中使用的许多细胞毒性药物都会抑制 DNA 复制并诱导肿瘤细胞的程序性细胞死亡（细胞凋亡）。 DNA复制本身的抑制程度尚不清楚 负责抗癌药物诱导细胞死亡。 细胞 用已知的抑制剂治疗可以诱导循环细胞死亡 DNA 复制的影响，例如 aphidicolin，一种抑制 DNA 复制的霉菌毒素 DNA聚合酶α和δ延长复制，或 含羞草碱，一种植物氨基酸，可阻止细胞复制的启动 哺乳动物细胞。该应用程序的主要目标是识别基因 以及参与诱导细胞死亡的遗传途径 通过DNA复制抑制剂。 这些基因将通过 遗传抑制元件 (GSE) 的表达选择，短 cDNA 通过编码显性作用抑制基因表达的片段 肽或抑制性反义RNA。 标准化的 GSE 库 逆转录病毒载体将被转导到HeLa细胞中，并且GSE 保护细胞免受阿非迪霉素或含羞草碱诱导的细胞死亡 孤立。这些 GSE 将以其功效、效果为特征 影响细胞周期进程、保护细胞免受不同 DNA 侵害的能力 复制抑制剂和细胞类型特异性。 全长 cDNA 与所选 GSE 相对应的人类基因将被分离，并且 它们的结构和在细胞周期中的表达模式将是 特点。 还将测试孤立的 GSE 的效果 关于细胞对复制抑制化疗药物的反应， 确定基于复制的细胞毒性机制的作用 他们的抗肿瘤作用。