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重复的单链悬垂物，该端子被提出，以帮助在染色体末端形成一个环结构。大量证据表明，通过实验破坏，DNA损伤或进行性端粒缩短发生端粒悬垂的暴露。可以通过添加与端粒3'悬垂同源的小型DNA寡核苷酸来模仿这种暴露，作为DNA损伤反应（例如凋亡）的信号。为了探索此反应的机制和可能的治疗效用，我们设计了与端粒悬垂物同源的11-碱基寡核苷酸，用于研究黑色素瘤中凋亡的研究。目的：（1）研究p73，e2f1和rb在端粒悬垂DNA引起的凋亡反应中的作用。 （2）检查抗凋亡蛋白livin的降低以及端粒悬垂DNA的激活。 （3）研究黑色素瘤细胞中黑色素细胞分化标记物的端粒悬垂序列，例如MART-1/MELAN A，酪氨酸酶，GP100，TRP-1和TRP-2。 （4）研究这些寡核苷酸是否会减少SCID小鼠中转移性结节的数量和肿瘤的大小。 各种蛋白质的调节和调节将通过蛋白质印迹研究。凋亡的分析将在FACS扫描上进行，所有转染将由Lipofectamine Plus进行，并通过Western印迹进行分析。 意义：上述项目将研究这些寡核苷酸在体内诱导凋亡的机制和实用性，及其对肿​​瘤性肿瘤性和转移的影响。高转移率对黑色素瘤的病原体造成了很大的影响，是黑色素瘤患者死亡的主要原因。该项目将促进这些基于端粒的DNA破坏模拟寡核苷酸作为一种新型的抗癌疗法的发展。