TELOMERASE ASSOCIATED FACTORS AND THEIR ROLES IN CANCER PROGRESSION

端粒酶相关因子及其在癌症进展中的作用

• 批准号: 7724195
• 负责人: ELIZABETH H BLACKBURN
• 金额: $ 0.81万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-06-01 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7724195
• 关键词: Affinity Chromatography; Biochemistry; Cancer Cell Growth; Cancer cell line; Cell Proliferation; Cells; Complex; Computer Retrieval of Information on Scientific Projects Database; DNA Microarray Chip; DNA Microarray format; Data; Disruption; Double-Stranded RNA; Funding; Gene Expression; Grant; Growth; HeLa S3; Human; Induction of Apoptosis; Institution; Interferons; Malignant Neoplasms; Mass Spectrum Analysis; Microarray Analysis; Proteins; Protocols documentation; Research; Research Personnel; Resources; Ribonucleoproteins; Role; Small Interfering RNA; Source; Structure; Telomerase; Telomerase RNA Component; Telomerase inhibition; Telomere Length Maintenance; Telomere Shortening; United States National Institutes of Health; cancer cell; cell growth; novel; response; telomere; tumor progression

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Cancer cells commonly upregulate the ribonucleoprotein-telomerase. Inhibition of telomerase has previously been predicted to slow cancer growth, but only after a sufficient period elapses for telomere shortening to occur. Recently, we have inhibited cancer cell growth specifically by inhibiting telomerase function through an siRNA targeting the template region of wild-type telomerase RNA. Expression of such siRNA in cancer cells resulted in dramatic reduction of telomerase activity, rapid inhibition of cell growth and induction of apoptosis. Unexpectedly, those potent cellular effects were not apparently induced by the disruption of telomere structures, suggesting that inhibition of telomerase by siRNA may inhibit a function of telomerase in cancer cells other than telomere length maintenance. DNA microarray analysis in these cells indicates rapid changes in growth-related gene expression that is independent of interferon response triggered by double stranded RNA. These data suggested that telomerase may be essential for cancer cell proliferation in addition to telomere length maintenance. Therefore, a main focus of our research is to further understand the underlying mechanisms by which telomerase regulates cell proliferation through identification of novel telomerase associated factors. Biochemistry purification of telomerase complexes from human cancer cell line Hela S3 has been achieved using Tandem affinity purification (TAP) protocol. Proteins associated specifically with telomerase will be isolated and identified by Mass Spectrometry.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 癌细胞通常上调核糖核蛋白端粒酶。此前曾预测抑制端粒酶可以减缓癌症的生长，但只有在端粒缩短发生足够长的时间后才会发生。最近，我们通过针对野生型端粒酶RNA模板区域的siRNA抑制端粒酶功能，从而特异性抑制癌细胞生长。此类 siRNA 在癌细胞中的表达导致端粒酶活性显着降低、细胞生长快速抑制并诱导细胞凋亡。出乎意料的是，这些有效的细胞效应显然不是由端粒结构的破坏引起的，这表明siRNA对端粒酶的抑制可能会抑制癌细胞中除端粒长度维持之外的端粒酶功能。这些细胞中的 DNA 微阵列分析表明，生长相关基因表达的快速变化与双链 RNA 触发的干扰素反应无关。这些数据表明，除了维持端粒长度之外，端粒酶可能对癌细胞增殖也至关重要。因此，我们研究的一个主要重点是通过鉴定新的端粒酶相关因子来进一步了解端粒酶调节细胞增殖的潜在机制。使用串联亲和纯化 (TAP) 方案对人癌细胞系 Hela S3 中的端粒酶复合物进行生物化学纯化。将通过质谱分离和鉴定与端粒酶特异性相关的蛋白质。