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.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 癌细胞通常上调核糖核蛋白 - 凝集素酶。先前预测端粒酶的抑制作用会减缓癌症的生长，但仅在经过足够的时期以供端粒缩短发生后。最近，我们通过针对野生型端粒酶RNA的模板区域抑制端粒酶功能特别抑制癌细胞生长。这种siRNA在癌细胞中的表达导致端粒酶活性急剧降低，快速抑制细胞生长和凋亡的诱导。出乎意料的是，这些有效的细胞作用显然不是由于端粒结构的破坏而引起的，这表明siRNA抑制端粒酶可能会抑制端粒酶在端粒长度维持以外的癌细胞中的功能。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，除端粒长度维持外，端粒酶除了维持端粒外，对于癌细胞的增殖至关重要。因此，我们研究的主要重点是进一步了解端粒酶通过鉴定新型端粒酶相关因素来调节细胞增殖的潜在机制。使用串联亲和力纯化（TAP）方案实现了人类癌细胞系HELA S3的端粒酶复合物的生物化学纯化。特异性与端粒酶相关的蛋白质将通过质谱分离和鉴定。