膀胱癌癌旁组织端粒酶活性检测的临床意义

目的　探讨膀胱癌癌旁组织端粒酶活性检测的临床意义。　方法　采用端粒重复序列扩增 (TRAP)法 ,检测 2 4例膀胱癌组织及癌旁组织中端粒酶活性表达。　结果　 2 4例癌旁组织中端粒酶活性表达阳性 10例 (42 % ) ,癌旁组织端粒酶活性表达与原发灶癌病理分级分期相关 ,并与癌复发相关。　结论　膀胱癌癌旁组织端粒酶活性检测可作为判断膀胱癌预后的指标之一     

端粒酶hTERT基因反义核酸对前列腺癌细胞PC3端粒酶活性的抑制作用

目的探讨hTERT基因的两端部分硫代修饰反义寡核苷酸（ASPS—ODN）对前列腺癌细胞PC3端粒酶活性的抑制作用。方法采用端粒重复序列扩增法及TRAP-PCR-ELISA法检测前列腺癌细胞的端粒酶活性；采用RT-PCR方法检测hTERT基因mRNA的表达水平；以免疫组化通过流式细胞仪检测hTERT基因蛋白水平的变化。结果hTERT基因的两端部分硫代修饰ASPS—ODN作用于PC3细胞48h，其端粒酶活性下降，作用72h，其端粒酶活性受到抑制。结论通过hTERT基因的两端部分硫代修饰ASPS—ODN特异性抑制hTERT基因mRNA的表达，可降低前列腺癌细胞PC3端粒酶活性。     

血管瘤组织中端粒酶逆转录酶基因表达与端粒酶活性的关系

目的　探讨端粒酶逆转录酶 (hTERT )基因在血管瘤组织中的表达及其与端粒酶活性的关系。方法　采用原位逆转录 聚合酶链反应 (RT PCR )和链霉抗生物素蛋白 过氧化物酶(SP)免疫组织化学方法分别检测 40例 (增生期 2 0例、退化期 2 0例 )血管瘤组织及 2 0例正常皮肤组织中hTERTmRNA表达 ,并采用端粒重复序列扩增 (TRAP)法测定血管瘤组织中端粒酶活性。结果　增生期血管瘤组织中hTERTmRNA阳性表达率为 80 .0 % ,退化期血管瘤组织中hTERTmRNA阳性表达率为 15 .0 % ,而正常皮肤组织中hTERTmRNA无阳性表达 ,增生期血管瘤与退化期血管瘤hTERTmRNA表达率差异具有显著性 (P <0 .0 1)。增生期血管瘤组织中端粒酶活性为81.4% ,退化期血管瘤组织中端粒酶活性 2 1.6% ,正常皮肤组织中无端粒酶活性 ,增生期血管瘤与退化期血管瘤组织中端粒酶活性比较 ,差异有显著性 (P <0 .0 1)。结论　端粒酶活性与血管瘤内皮细胞增殖状况和hTERTmRNA表达密切相关 ,在血管瘤的发生和发展中起着重要的作用。     

表没食子儿茶素没食子酸酯对肝癌细胞端粒酶逆转录酶和c-myc基因表达的调控作用

目的探讨表没食子儿茶素没食子酸酯(EGCG)作用肝癌细胞时端粒酶活性的变化、端粒酶逆转录酶(TERT)及cmyc基因表达的改变,以探讨EGCG对肝癌细胞端粒酶活性的调控机制。方法用聚合酶链反应免疫酶联吸附试验(PCR-ELISA)测定肝癌细胞端粒酶活性,逆转录聚合酶链反应(RTPCR)检测肝癌细胞中人端粒酶逆转录酶(hTERT)和c-mycmRNA表达。结果在一定时间、一定剂量范围内,EGCG能抑制肝癌细胞端粒酶活性,随着浓度的增加和时间的延长,端粒酶活性逐渐下调;尤其当EGCG浓度超过50mg/L或作用时间超过36h时,端粒酶活性下调更明显,与对照组比较差异用统计学意义(P<0.01);EGCG能抑制肝癌细胞hTRETmRNA的表达,其下降趋势与端粒酶活性下降趋势基本一致,且下调幅度较端粒酶活性下降更明显,两者呈正相关(r=0.931,P<0.01);而EGCG对肝癌细胞cmycmRNA表达的抑制率比hTRETmRNA表达的抑制率更高,并且c-mycmRNA表达先受到抑制,与hTRETmRNA表达的抑制相关明显(r=0.907,P<0.01)。结论EGCG对c-myc的抑制作用可能导致hTERTmRNA的抑制并进一步下调端粒酶活性。     

The association between telomerase, histopathological parameters, and KI-67 expression in breast cancer.

BACKGROUND: Telomerase is a ribonucleoprotein enzyme that appears to play an important role in carcinogenesis. Telomerase reactivation seems to be associated with immortalization and malignancy. METHODS: Using a polymerase chain reaction (PCR)-based assay known as the TRAP (telomeric repeat and amplification protocol) assay, we examined telomerase activity in 60 breast specimens prospectively collected from 39 patients undergoing elective breast surgery in our center. The specimens included adjacent noncancerous breast (n = 21), benign breast disease (n = 5), and infiltrating carcinoma (n = 34). Ki-67 expression was determined in 32 invasive breast cancer specimens using immunohistochemistry techniques. The histopathological features were determined by light microscopy by an experienced breast pathologist. RESULTS: Telomerase activity was detected in 24 (71%) of 34 infiltrating carcinomas. None of the adjacent noncancerous specimens nor the benign breast lesions expressed telomerase activity. Telomerase reactivation was significantly associated with nodal metastasis and Ki-67 expression. There was no significant association between telomerase activity and menopausal status, tumor grade, or tumor size. CONCLUSIONS: Telomerase reactivation is associated with the acquisition of malignancy in the human breast. Telomerase activity is significantly associated with nodal metastasis and cellular proliferation as measured by Ki-67 expression in human breast cancer.     

Detection of telomerase activity in breast masses by fine-needle aspiration

Background: Telomerase is an RNA-dependent DNA polymerase that compensates for the telomere shortening that occurs in its absence. Reactivation of telomerase is thought to be an important step in cellular immortalization, and recent studies have indicated that telomerase activity is often detected in primary human malignancies. The clinical implications of telomerase activity in human tumors are currently under investigation. Methods: Eighty-nine samples (46 FNAs and 43 gross tissue biopsies) from 44 patients with breast masses were analyzed prospectively for the presence of telomerase activity by a modification of the telomere repeat amplification protocol (TRAP). All samples were obtained directly from the excised mass at the time of specimen removal in the operating room. Results: Telomerase activity was detected in 17 of 19 (90%) FNA samples and 15 of 18 (83%) invasive breast cancer tissue biopsies. Telomerase was also detected in 9 of 16 (56%) FNAs and 8 of 15 (53%) tissue biopsies from 16 fibroadenomas. Other benign proliferative lesions (n=5) did not have detectable telomerase activity in either FNA or tissue specimens. FNA-TRAP results correlated with the gross tissue specimen TRAP results in 95% of all cases. Conclusion: The FNA-TRAP assay for telomerase detection is a highly sensitive and accurate method for the detection of telomerase activity in breast masses. Future application of these techniques should facilitate evaluation of telomerase as a tumor marker in the clinical management of breast and other solid malignancies. These authors contributed equally to this work.     

PCR-ELISA法检测膀胱肿瘤患者尿脱落细胞端粒酶活性

目的：探讨尿脱落细胞端粒酶活性变化在膀胱肿瘤诊断中的作用。方法应用PCR－ELISA法检测53例膀胱肿瘤患者尿液脱落细胞端粒酶的活性。结果：非膀胱肿瘤和膀胱肿瘤患者尿液脱落细胞端粒酶活性阳性率分别为64．15％（34．53）和7．69％（2／26）,健康对照者7例均为阴性,膀胱肿瘤患者与正常人及非膀胱肿瘤患者的端粒酶活性分别相比,差别均有极显著性意义（P＜0．001）。但端粒酶活性与肿瘤的分期分级无相关性。结论：尿脱落细胞端粒酶活性检测可以作为诊断膀胱肿瘤的无创性检测方法,但不能预测膀胱肿瘤的临床分期分级。     

TRAP银染法检测端粒酶在胃癌组织中的表达

目的了解胃癌组织中端粒酶表达的状况及其临床意义,研究非放性 TRAP 检测端粒酶的可行性。方法用 TRAP 银染法检测58例胃癌及相应患者的正常胃粘膜,12例胃腺瘤和9例胃溃疡组织的端粒酶表达情况。结果 58例胃癌组织中49(84.5％)例端粒酶表达阳性,正常粘膜无一例检测到端粒酶活性(P<0.001),12例胃腺瘤,9例胃溃疡组织中各有1例检测到端粒酶活性。胃癌组织端粒酶的表达与年龄、肿瘤大小、分化程度、浸润深度、淋巴结转移及 TNM 分期等临床病理参数无明显相关(P<0.05)。结论端粒酶是一种较理想的胃癌肿瘤标记物,TRAP 银染法是一种值得推广的检测端粒酶活性的方法。     

膀胱癌尿脱落细胞端粒酶活性检测及其临床意义

目的检测尿脱落细胞端粒酶活性并探讨其临床意义。方法应用改良的端粒重复序列扩增（ＴＲＡＰ）银染方法,分别对膀胱癌组织、正常膀胱组织,以及膀胱癌患者和非尿路上皮肿瘤患者的尿脱落细胞、膀胱冲洗液进行端粒酶活性检测。结果１２例正常膀胱组织均无端粒酶活性,４８例膀胱癌组织中４４例（９１．７％）端粒酶阳性。膀胱癌患者尿液及膀胱冲洗液中脱落细胞端粒酶阳性率分别为８３．３％（４０／４８）和８７．５％（４２／４８）。１２例分化良好（Ｇ１级）膀胱癌患者中,尿液和膀胱冲洗液中脱落细胞端粒酶阳性率分别为７５．０％（９／１２）和８３．３％（１０／１２）。结论尿脱落细胞端粒酶活性检测敏感性高,可用于膀胱癌的早期诊断和术后随访。     

大肠癌肝转移肿瘤中端粒酶活性和p16基因纯合缺失

目的 研究人大肠癌肝转移肿瘤中端粒酶活性和p16基因纯合缺失及其临床意义。方法 采用TRAP银染方法和半定量多重PCR检测24例大肠癌转移肿瘤组织中端粒酶活性及p16基因的纯合缺失情况,并结合临床病理参数,进行统计学分析。结果 本组24例大肠癌肝转移肿瘤组织中检测到19例端粒酶阳性,阳性率为79．2％。端粒酶活性与转移瘤大小、肝内转移灶数目、分化程度、HBsAg以及是否伴有纤维化无关。在24例转移瘤组织中有9例标本中检测到p16基因的缺失,阳性率为37．5％,p16基因的失与端粒酶活性相关。结论 大肠癌肝转移肿瘤中端粒酶活性和p16基因异常对阐明大肠癌转移的生物学行为有重要的意义。     

胰液中端粒酶及亚单位检测对胰腺癌的诊断价值

目的　探讨检测胰液中端粒酶逆转录酶 (hTERT)mRNA表达和端粒酶活性对胰腺癌的诊断与鉴别诊断价值。方法　对比分析 2 4例胰腺癌和 14例慢性胰腺炎患者胰液中的hTERTmRNA和端粒酶活性的检测结果。结果　hTERTmRNA在胰腺癌中表达阳性率为 87.5 % ( 2 1/ 2 4) ;慢性胰腺炎中阳性率为 2 1.4% ( 3 / 14 ) (P <0 .0 0 1)。胰腺癌与慢性胰腺炎胰液中端粒酶阳性检出率分别为83 .3 % ( 2 0 / 2 4) ,2 8.6% ( 4 / 14 ) (P <0 .0 0 1)。结论　胰液中检测hTERTmRNA表达和端粒酶活性对胰腺癌诊断与鉴别诊断有一定价值。