The Effect of the Telomere 3' Overhang DNA on Melanoma.

端粒 3 突出 DNA 对黑色素瘤的影响。

• 批准号: 6795699
• 负责人: NEELU PURI
• 金额: $ 7.63万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-15 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6795699
• 关键词: DNA; DNA damage; SCID mouse; SDS polyacrylamide gel electrophoresis; apoptosis; biological signal transduction; cell differentiation; cell growth regulation; cysteine endopeptidases; flow cytometry; melanoma; metastasis; neoplasm /cancer therapy; nucleic acid sequence; oligonucleotides; pathologic process; telomere; terminal nick end labeling; transfection; western blottings

DESCRIPTION (provided by applicant): The 3' end of each telomere is a single-stranded overhang of repeats of TTAGGG that is proposed to help form a loop structure at the chromosome ends. Considerable evidence suggests the exposure of the telomere overhang occurs by experimental disruption, DNA damage or progressive telomere shortening. This exposure can be mimicked by adding small DNA oligonucleotides homologous to the telomere 3' overhang as a signal for DNA damage responses such as apoptosis. To explore the mechanism and possible therapeutic utility of this response we designed an 11-base oligonucleotide homologous to the telomere overhang for studies on apoptosis in melanoma described below. Aims: (1) Study the role of p73, E2F1 and Rb on the apoptotic response induced by the telomere overhang DNA. (2) Examine the decrease of anti-apoptotic protein livin and activation of caspases by telomere overhang DNA. (3) Study the induction by the telomere overhang sequence of markers of melanocyte differentiation such as Mart-1/Melan A, Tyrosinase, gp100, TRP-1 and TRP-2 in melanoma cells. (4) Investigate whether these oligonucleotides will reduce the number of metastatic nodules and size of tumors in SCID mice. The modulation and regulation of the various proteins will be studied by Western blotting. Analysis of apoptosis will be done on the FACS scan and all transfections will be performed by lipofectamine plus and analysed by Western blotting. Significance: The above project will study the mechanism and utility of these oligonucleotides to induce apoptosis in vivo and in vitro and its effect on tumodgenecity and metastasis of tumors.The high rate of metastasis contributes greatly to the pathogenesis of melanoma and are major causes of death among melanoma patients. The project will facilitate the development of these telomere based DNA-damage mimetic oligonucleotides as a novel anti-cancer therapy.

描述（由申请人提供）：每个端粒的 3' 末端是 TTAGGG 重复的单链突出端，建议帮助在染色体末端形成环结构。大量证据表明，端粒突出的暴露是由于实验破坏、DNA 损伤或进行性端粒缩短而发生的。这种暴露可以通过添加与端粒 3' 突出端同源的小 DNA 寡核苷酸来模拟，作为 DNA 损伤反应（例如细胞凋亡）的信号。为了探索这种反应的机制和可能的治疗效用，我们设计了一种与端粒突出端同源的 11 碱基寡核苷酸，用于下述黑色素瘤细胞凋亡的研究。目的：(1)研究p73、E2F1和Rb在端粒突出DNA诱导的细胞凋亡反应中的作用。 (2)检查端粒突出DNA对抗凋亡蛋白livin的降低和caspases的激活。 (3)研究黑色素细胞分化标志物Mart-1/Melan A、酪氨酸酶、gp100、TRP-1和TRP-2等端粒突出序列对黑色素瘤细胞的诱导作用。 (4) 研究这些寡核苷酸是否会减少 SCID 小鼠中转移结节的数量和肿瘤的大小。 将通过蛋白质印迹研究各种蛋白质的调节和调节。细胞凋亡分析将在 FACS 扫描上进行，所有转染将通过 lipofectamine plus 进行并通过蛋白质印迹进行分析。 意义：该项目将研究这些寡核苷酸在体内外诱导细胞凋亡的机制和用途，以及其对肿瘤发生和转移的影响。高转移率对黑色素瘤的发病有重要影响，是导致死亡的主要原因在黑色素瘤患者中。该项目将促进这些基于端粒的 DNA 损伤模拟寡核苷酸的开发，作为一种新型抗癌疗法。