Cloning and clinical application of telomerase cDNA

端粒酶cDNA的克隆及临床应用

• 批准号: 09557195
• 负责人: IDE Toshinori
• 金额: $ 8.64万
• 依托单位: HIROSHIMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09557195/
• 关键词: telomere; telomerase; cellular lifespan; cellular senescence; cellular immortality; cancer diagnosis; テロメラーゼcDNA; hTRT; テロメラーゼ発現; 細胞癌化; 癌組織

New findings of this project are 1) telomerase was partially purified from porcine testis to a few protein bands on acrylamidegel electrophoresis, but, since cDNA (hTERT) cloning of assumed catalytic subunit of telomerase has been reported by other reserchers (Aug. 1 997), we abandoned cloning project through protein, purification, 2) expression of hTERT mRNA was found to be correlated with presence 0f telomerase activity, 3) telomerase-negative human fibroblasts expressed telomerase activity after transfection with hTERT cDNA, 4) these telomerase positive human fibroblasts extended both telomere length and proliferative lifespan, but they did not immortalized, 5) telomerase-negative human fibroblasts transformed with SV40 T-antigen also expressed telomerase activity after. transfection with hTERT cDNA, 6) these telomerase-positive SV4O-transformed human fibroblasts extended both telomere length and proliferative lifespan, and they immortalized.

本项目的新发现有：1)从猪睾丸中部分纯化到端粒酶，在丙烯酰胺凝胶电泳上显示出几个蛋白带，但由于其他研究者已经报道了端粒酶催化亚基的cDNA （hTERT）克隆（1997年8月），我们放弃了通过蛋白纯化的克隆项目；2)发现hTERT mRNA的表达与端粒酶活性存在相关。3)转染hTERT cDNA后，端粒酶阴性的人成纤维细胞表达端粒酶活性；4)端粒酶阳性的人成纤维细胞延长了端粒长度和增殖寿命，但没有长生不老；5)转染SV40 t抗原后，端粒酶阴性的人成纤维细胞也表达了端粒酶活性。转染hTERT cDNA后，这些端粒酶阳性的sv40o转化的人成纤维细胞延长了端粒长度和增殖寿命，并实现了长生不老。

项目成果

Hiyama,T.: "In situ mRNA hybridizaion technique for analysis of human telomerase RNA in gastric precancerous and cancerous lesions." Jpn.J.Cancer Res.89. 1099-1103 (1998)

Hiyama,T.：“用于分析胃癌前期和癌性病变中人端粒酶 RNA 的原位 mRNA 杂交技术。”

Yasui, W., et al.: "Expression of telomerase catalytic component, telomerase reverse transcriptase, in human gastric carcinoma." Jpn.J.Cancer Res.89. 1187-1194 (1998)

Yasui, W., et al.：“端粒酶催化成分、端粒酶逆转录酶在人胃癌中的表达。”

Yoshiki,T.: "Extra-Chromosomal telomerase repeat DNA in telomerase-negative immortalized cell lines." Biochem.Biophys.Res.Com.247. 765-772 (1998)

Yoshiki,T.：“端粒酶阴性永生化细胞系中的染色体外端粒酶重复 DNA。”

Kitamoto, M., et al.: "Telomerase activity in precancerous hepatic nodules." Cancer. 85. 245-246 (1999)

Kitamoto, M., et al.：“癌前肝结节中的端粒酶活性。”

Hirose, M., et al.: "A new method to detect and measure telomerase activity by transcription-mediated amplification (TMA) and hybridization protection assay (HPA)." Clinical Chem.44. 2446-2452 (1998)

Hirose, M. 等人：“一种通过转录介导的扩增 (TMA) 和杂交保护测定 (HPA) 检测和测量端粒酶活性的新方法。”