TARP1在端粒酶调控和乳腺癌发生中的作用研究

Telomere length and telomerase activity are directly relevant to human aging, a variety of diseases and tumorgenesis. Telomerase binding proteins play an important role in the telomere and telomerase regulation. Recently, we applied Bimolecular Fluorescence complementation technology (BiFC) to do high throughput screening of TERT interacting protein and found a novel telomerase binding protein TARP1, which can interact with core components of telomerase including TERT, DKC1 amd TPP1. We hypothesized that TARP1 acts as a central linker protein for telomerase complex and plays an important role in telomerase regulation. With the support of this project, firstly we will use biochemistrical and molecular biological methods to reveal the function and mechanism of TARP1 on telomerase regulation in human breast cancer cell lines. Then we will use BiFC system to screen and identify TARP1 interacting proteins. Through immunohistochemical study of tumor biopsy from breast cancer patients, we will analyse relationship between expression of the TARP1 and key breast cancer tumor markers. Furthermore，we will use TARP1-knockout mice and TARP1 knockout breast cancer cell lines in combined with the tumorgenesis experiments in nude mice, to study the the function and mechanism of TARP1 in breast cancer carcinogenesis in vivo. This research will not only greatly promote our understanding of role of TARP1 protein in telomerase regulation and telomere maintaining, but also provide new insight to relationship between telomerase assembly control and tumorgenesis.

端粒的长度和端粒酶活性与人类的衰老、疾病及肿瘤发生密切相关，端粒酶结合蛋白起着调控端粒和端粒酶的作用。在近期筛选端粒酶结合蛋白的研究工作中我们最新发现， TARP1蛋白能与端粒酶复合物核心组分TERT、DKC1以及TPP1相互作用。我们推测TARP1可能作为端粒酶复合物组分的连接蛋白，在端粒酶调控中起重要作用。申请人拟在本项目支持下，首先利用生化、分子生物学手段在细胞水平揭示TARP1在端粒酶调控中的功能和机制；然后运用双分子荧光互补技术筛选、鉴定与TARP1互作的蛋白；进一步利用乳腺癌病人的病理切片，通过免疫组化分析TARP1表达及其与乳腺癌关键指标的关系，同时制备TARP1敲除乳腺癌细胞系，并结合TARP1基因敲除小鼠乳腺癌模型深入研究其在乳腺癌发生中的作用和机制。本研究将推进对TARP1蛋白在端粒酶调控中作用的了解，并为深入理解端粒酶组装调控和肿瘤发生的关系提供新视野.

端粒的长度和端粒酶活性与人类的衰老、疾病及肿瘤发生密切相关，端粒酶结合蛋白起着调控端粒和端粒酶的作用。在BiFC筛选端粒酶结合蛋白的研究工作中我们发现 TARP1蛋白能与端粒酶复合物核心组分TERT、DKC1以及TPP1相互作用。我们推测TARP1可能作为端粒酶复合物组分的连接蛋白，在端粒酶调控中起重要作用。我们首先利用生化、分子生物学手段在细胞水平揭示TARP1在端粒酶调控中的功能和机制；然后运用双分子荧光互补技术筛选、鉴定与TARP1互作的蛋白；进一步利用乳腺癌病人的病理切片，通过免疫组化分析TARP1表达及其与乳腺癌关键指标的关系，同时制备TARP1敲除乳腺癌细胞系，并制备TARP1基因敲除小鼠研究其在端粒酶调控和乳腺癌发生中的作用和机制。本研究探索了TARP1蛋白以及其他端粒相关蛋白包括TOE1、DTX2等在端粒酶调控中作用，并为深入理解端粒酶组装调控和肿瘤发生的关系提供新视野。