无需PCR扩增端粒酶检测荧光法的研究

Telomerase can synthesize telomere to maintain the length and structural integrity of telomere DNA using its RNA as a template, which can be found inside of 85% cancer cell. As one of the most important biomarker, it is of significant importance for early diagnosis of cancer to develop ultra-sensitive and high specific methods for detection of telomerase. New strategies for signal amplification and design of molecular recognition probe will be systematically studied based on the elongation mechanism of telomere DNA by telomerase using telomerase RNA as a template. Realtime detection of telomerase under physiological conditions will be explored by combing the high specificity of hairpin probe and signal amplification of nanoprobe and enzyme. New mechanism and methods will be established for simple and fast determination of telomerase without PCR technique. The inhibitor with potential anti-cancer ability will be screened by investigating the inhibition of G-quadruplex ligands on the telomerase activity. Therefore, it is of great theoretical and practical importance to the cancer early diagnosis and design and screening of anti-tumor drugs.

端粒酶依靠自身RNA模板合成端粒DNA、维持端粒的长度与结构完整，使得肿瘤细胞无限增殖，在85%的恶性肿瘤细胞内存在端粒酶。因此，作为诊断恶性肿瘤的重要标示物之一，端粒酶的超灵敏、高特异性检测对于恶性肿瘤的早期诊断具有极其重要的意义。本项目拟基于端粒酶以自身RNA为模板延长端粒DNA的机理，系统深入地研究分子识别探针设计和信号放大新策略。利用发夹型核酸探针的高特异性和纳米复合探针及酶辅助信号放大作用，探索近生理条件下实时检测端粒酶活性的新途径，提出研究端粒酶活性和抑制的新原理，构建无需PCR扩增特异性检测端粒酶的新方法。考察G-四链体配体对端粒酶活性的抑制，筛选具有抗肿瘤潜能的端粒酶抑制剂。此项目研究对于恶性肿瘤的早期诊断和抗肿瘤药物的设计与筛选具有重要的理论和实际意义。