EFFECTS OF ANTIMETABOLITES ON RNA METABOLISM

抗代谢药对 RNA 代谢的影响

• 批准号: 3172016
• 负责人: BRUCE JEFFREY DOLNICK
• 金额: $ 7.9万
• 依托单位: ROSWELL PARK CANCER INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-05-01 至 1993-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3172016
• 关键词: DNA replication; RNA biosynthesis; RNA splicing; antimetabolites; antineoplastics; cell bank /registry; cell type; dihydrofolate reductase; drug metabolism; drug resistance; floxuridine; fluorouracil; human tissue; messenger RNA; methotrexate; neoplasm /cancer pharmacology; nucleic acid metabolism; polymerase chain reaction; precursor mRNA; ribosomal RNA

The purpose of this application is to extend previous studies of the effects of 5-fluorouracil (FUra) on RNA metabolism in an effort to better understand the RNA mediated effects of FUra cytotoxicity. This study will focus on two FUra induced aberrations of mRNA metabolism, pre-mRNA splicing and mRNA half life. It is specifically proposed that the following be accomplished: 1. Apply the polymerase chain reaction to quantitate steady state levels of dihydrofolate reductase (DHFR) RNA splicing substrates as well as splicing intermediates in the pre-mRNA processing pathway, in FUra treated and control cells. Extend this assay to non-gene amplified cells so that it may eventually be applied in vivo. 2. Measure the half life of 3.5 kb DHFR mRNA in cells grown in the presence or absence of FUra. Assess whether an increased half-life after cell growth in medium containing FUra is an explanation for the previously observed increase in this specific DHFR mRNA. Identify the 3' noncoding element(s) present in 3.5 kb DHFR mRNA which appear to mediate this effect of FUra on half-life deletion mapping and DNA transfection. 3. Study the effect of FUra pretreatment on methotrexate cytotoxicity in cells expressing mRNAs with altered 3' noncoding regions to correlate alterations in mRNA half life with DHFR drug responsiveness.

该应用的目的是扩展以前的研究 5-氟尿嘧啶（Fura）对RNA代谢的影响，以改善 了解RNA介导的Fura细胞毒性的作用。 这项研究会 专注于两个Fura诱导的mRNA代谢畸变，前MRNA剪接 和mRNA半生活。 特别提出以下是 完成： 1。应用聚合酶链反应来定量稳态水平 二氢叶酸还原酶（DHFR）RNA剪接底物以及 在Fura处理的MRNA处理途径中的剪接中间体 和控制细胞。 将此测定扩展到非基因扩增的细胞，以便 它最终可以在体内应用。 2。测量在生长在细胞中的3.5 kb DHFR mRNA的半寿命 存在或不存在Fura。 评估是否增加了半衰期之后 含有Fura的培养基中的细胞生长是对先前的 观察到该特定DHFR mRNA的增加。 识别3'非编码 3.5 kb DHFR mRNA中存在的元素似乎介导了这种效果 半衰期缺失映射和DNA转染的Fura。 3。研究FURA预处理对甲氨蝶呤细胞毒性的影响 用改变的3'非编码区域表达mRNA的细胞以相关 DHFR药物反应能力的mRNA半衰期改变。

项目成果

Quantitation of dihydrofolate reductase and thymidylate synthase mRNAs in vivo and in vitro by polymerase chain reaction.

通过聚合酶链反应对体内和体外二氢叶酸还原酶和胸苷酸合酶 mRNA 进行定量。

• 发表时间: 1992
• 期刊: Oncology research
• 影响因子: 3.1
• 作者: Dolnick,BJ;Zhang,ZG;Hines,JD;Rustum,YM
• 通讯作者: Rustum,YM

5-Fluorouracil substitution alters pre-mRNA splicing in vitro.

• DOI: 10.1016/s0021-9258(18)68949-5
• 发表时间: 1988-03
• 期刊: The Journal of biological chemistry
• 作者: S. Doong;B. Dolnick

5-Fluorouracil augmentation of dihydrofolate reductase gene transcripts containing intervening sequences in methotrexate-resistant KB cells.

5-氟尿嘧啶增强甲氨蝶呤耐药 KB 细胞中含有插入序列的二氢叶酸还原酶基因转录本。

• 发表时间: 1986
• 期刊: Molecular pharmacology
• 影响因子: 3.6
• 作者: Will,CL;Dolnick,BJ
• 通讯作者: Dolnick,BJ

5-Fluorouracil alters dihydrofolate reductase pre-mRNA splicing as determined by quantitative polymerase chain reaction.

通过定量聚合酶链式反应测定，5-Fluorouracil 会改变二氢叶酸还原酶前体 mRNA 剪接。

• 发表时间: 1993
• 期刊: Molecular pharmacology
• 影响因子: 3.6
• 作者: Wu,XP;Dolnick,BJ
• 通讯作者: Dolnick,BJ

5-Fluorouracil inhibits dihydrofolate reductase precursor mRNA processing and/or nuclear mRNA stability in methotrexate-resistant KB cells.

5-Fluorouracil 抑制甲氨蝶呤耐药 KB 细胞中二氢叶酸还原酶前体 mRNA 加工和/或核 mRNA 稳定性。

• 发表时间: 1989
• 期刊: The Journal of biological chemistry
• 作者: Will,CL;Dolnick,BJ
• 通讯作者: Dolnick,BJ