STUDY OF TELOMERASE ACTIVITY AND EXPRESSION OF THE CATALYTIC SUBUNIT ASSOCIATED WITH THIS GENE IN HUMAN UROLOGICAL CANCER.

研究人类泌尿癌中端粒酶活性和与该基因相关的催化亚基的表达。

• 批准号: 09671645
• 负责人: FUJIOKA Tomoaki
• 金额: $ 1.86万
• 依托单位: IWATE MEDICAL UNIVERSITY SCHOOL OF MEDICINE,DEPARTMENT OF UROLOGY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 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-09671645/
• 关键词: Telomerase activity; Telomerase-subunit; TRAP assay; RT-PCR method; renal cell carcinoma; bladder cancer; oncogene; tumor-maker; テロメラーゼ; 分子生物学; テロメラーゼ・サブセット; hEST2; hTRT; TLP1; TPI; 膀胱正常組織; 集団検診; 尿剥離細胞; TRAP Assay; 前立腺癌; テロメア長; サザンブロツ

Teromerase is the ribonucleoprotein enzyme associated with the immortalization and oncogenesis of cancer cells. Telomerase activity was examined in 47 RCCs using PCR-based telomeric repeat amplification protocol (TRAP) assay. Among 30 RCCs, 18 samples(60%) displayed the activity, white none of the normal tissues exhibited the activity. Subdivision of the tumors according to telomerase activity did not reveal any obvious difference in distribitution of tumor size, stage, histocytological subtype, or DNA-ploidy. However, statistically significant relationship between the frequency of telomerase-positive activity and serum lAP levels or tumor grade was foumd, respectively (p<0.05). These results indicate that telomerase activity may be an important factor for' evaluating the malignancy potential of RCC, rather than for detecting malignancy of the kidney.Although TRAP assay is a highly sensitivity method, it has been reported that Taq inhibitor and other factors in method might cause false results during PCR amplification. Recently, three new major components of telomerase genes which are hTR, TLP1/TPI, and hEST2/hTRT has been cloned. In 27 human urinary bladder cancers, expression of hEST2/hTRT and TLP1/TPI was studied by using revel-se transcriptase-polymerase chain reaction (RT-PCR) method. In all 27 cancers (100%), hEST2/hTRT expression was detected, while TLP1/TPL was displayed in 25 of 27 speciniens(92%). Five normal bladder tissues showed no expression of hEST2/hTRT, but 3 of 5 tisses (60%) did TLPl/TP1 expression. These results indicated that RT-PCR for detecting expression of hEST2/hTRP gene may be a powerful method for the mass screening or diagnosis of urinary bladder.

端粒酶是一种核糖核蛋白酶，与癌细胞的永生化和癌变有关。采用基于pcr的端粒重复扩增方法（TRAP）检测47例rcc的端粒酶活性。在30例rcc中，18例（60%）显示活性，其余正常组织均无活性。根据端粒酶活性对肿瘤进行细分，在肿瘤大小、分期、组织细胞学亚型、dna倍性分布等方面均无明显差异。端粒酶阳性活性频率与血清lAP水平、肿瘤分级有统计学意义（p<0.05）。这些结果表明，端粒酶活性可能是评估肾癌恶性潜能的重要因素，而不是检测肾脏恶性肿瘤的重要因素。虽然TRAP检测是一种高灵敏度的方法，但有报道称，方法中的Taq抑制剂等因素可能导致PCR扩增过程中出现错误结果。近年来，端粒酶基因的三个新的主要组成部分hTR、TLP1/TPI和hEST2/hTRT被克隆出来。采用逆转录-聚合酶链反应（rev -se transcripase -polymerase chain reaction, RT-PCR）方法对27例人膀胱癌组织中hEST2/hTRT和TLP1/TPI的表达进行了研究。在所有27例肿瘤中（100%）检测到hEST2/hTRT表达，而在27例肿瘤中有25例（92%）检测到TLP1/TPL表达。5例正常膀胱组织未见hEST2/hTRT表达，而5例组织中有3例（60%）有TLPl/TP1表达。以上结果提示，检测hEST2/hTRP基因表达的RT-PCR技术可作为大规模筛查或诊断膀胱疾病的有力手段。