肺癌组织的粒酶活性表达及意义

应用ＰＣＲ－端粒重复旬扩增法（ＴＲＡＰ）检测３１例肺癌、４例癌旁及２６例肺良性肿瘤患者肿瘤组织的端粒酶活性表达。结果显示，肺癌组织中端粒酶生表达率为８０．６％（２５／３１）、癌旁组织无表达、肺良性肿瘤组织表达率为７．７％（２／２６）。端粒酶活性表达与肺癌分化程度呈负相关，与肺癌分期无关。有及无淋巴结锗 的端粒酶活性表达率有显著性差异。提示端粒酶在肺癌的发生、发展及转移过程中皆起重要作用；其在肺癌组     

肺癌组织端粒酶活性测定的临床意义

目的：探讨端粒酶活性与肺癌的关系。方法：采用以聚合酶链反应（PCR）为基础的端粒重复序列扩增法（TRAP），结合聚丙酰胺凝胶电泳银染法检测经纤支气管镜取出的35例肺癌患者肺癌组织和相应健侧肺支气管粘膜及35例支气管－肺良性疾病患者支气管粘膜的端粒酶活性。结果：（1）肺癌组肺癌组织活检标本端粒酶阳性33例（94％），毛刷标本阳性32例（91％），其相应健侧肺活检标本阳性2例（5．7％），毛刷标本阳性4例（11％）；（2）支气管－肺良性疾病患者活检阳性0例（0％），毛刷阳性2例（5．7％）；（3）肺癌组织端粒酶活性与肺癌组织学类型和临床分期无明显关系（P＞0．05）。结论：端粒酶活性与肺癌密切相关，可作为检测肺组织癌的一种辅助手段。     

支气管活检标本的端粒酶活性检测对肺癌的诊断价值

目的：探讨肺癌组织端粒酶活性检测对肺癌的诊断价值。方法采用端粒酶重复序列扩增法检测纤维支气管镜活组织检查的肺癌病变组织（７０例）、肺癌对侧支气管组织（７０例）及非癌性肺疾病支气管组织（２０例）的端粒酶活性,并和病理学及细胞学检查结果进行比较。结果肺癌病变组织端粒酶检出率为８１％,明显高于肺癌对侧支气管组织（１４％）（Ｐ〈０．０１）及非癌性肺疾病支气管组织（１０％）的检出率（Ｐ〈０．０１）;肺癌组织     

外周血端粒酶活性对肺癌血转移诊断的探讨

目的：探讨以端粒酶重复序列扩增法(PCR—TRAP)检测血中肺癌端粒酶活性对于肺癌血转移诊断、分型分期的临床意义。方法：选取24例肺癌、26例肺良性疾病患，应用PCR—TRAP法测定端粒酶活性，磁分离酶免法检查癌胚抗原(CEA)及血沉。结果：肺癌组端粒酶活性阳性率明显高于肺良性疾病组(p＜0．01)，按肺癌恶化程度分，Ⅲ-Ⅳ期比Ⅰ-Ⅱ期高。CEA和血沉在肺癌与肺良性疾病组异常率无显性差异(p＞0．05)。22．2％(4／18)晚期肺癌外周血端粒酶活性呈阴性，肺良性疾病患中19．2％(5／26)外周血端粒酶活性呈阳性，肺癌患外周血端粒酶活性敏感性比CEA、ESR高(p＜0．01和p＜0．05)，特异性无显性差别(p＞0．05)。结论：端粒酶对于肺癌分期分型及预后判断有临床应用价值。改进检测方法，对提高肺癌血转移诊断的特异性和敏感度显得重要。     

肺癌支气管肺泡灌洗液细胞端粒酶测定的临床意义

目的探讨端粒酶活性在肺癌支气管肺泡灌洗液（ＢＡＬＦ）细胞中的表达与肺癌发生机制的关系。方法应用银染多聚酶链方法对肺癌患者和支气管、肺良性疾病患者各３０例的ＢＡＬＦ细胞端粒酶活性的表达进行检测。结果肺癌患者ＢＡＬＦ细胞端粒酶活性８３％阳性表达，支气管、肺良性疾病ＢＡＬＦ细胞表达均阴性（Ｐ＜０．００１）。同一肺癌患者患侧肺端粒酶活性在ＢＡＬＦ细胞８３％表达，健侧肺阳性表达２７％。结论端粒酶活性是肺癌诊断的标志物之一，与肺癌的发生发展密切相关。     

肺癌前病变、肺癌中FHIT基因表达的临床研究

背景与目的FHIT基因为近年发现的新候选抑癌基因，位于3P14．2跨越FRA3B易脆点，在包括肺癌在内的人类多种肿瘤中均存在异常表达。本研究旨在观察FHIT基因在人肺癌前病变、肺癌中表达情况，探讨FHIT基因在人肺癌发生、发展过程中的可能作用。方法采用免疫组化方法检测298例甲醛固定、石蜡包埋的标本（包括161例肺癌、51例肺癌前病变、30例正常肺组织、23例肺良性病变和33例肺癌转移淋巴结）中FHIT蛋白表达情况。结果FHIT蛋白在正常肺组织及肺良性病变组织中均无失表达；癌前病变组织及肺癌组织中失表达率分别为54．9％（28／51）和59．0％（95／161）：肺癌转移淋巴结组织中FHIT蛋白失表达率78．8％（26／33），各组问比较有显著性差异（P〈0．05）。肺癌组织中FHIT基因表达水平与肺癌组织学类型、肿瘤细胞分化程度、患者P-TNM分期、淋巴结转移程度存在相关性（P〈0．05）。FHIT蛋白失表达组肺癌患者的术后五年生存率显著低于表达组（P〈0．01）。吸烟组患者FHIT基因失表达率69．1％（94／136）显著高于无吸烟组49．5％（49／99）（P〈0．01）。结论FHIT蛋白失表达可能是肺癌发生过程中的早期分子事件，与肺癌的发生、发展及预后有关；吸烟导致FHIT蛋白表达下降可能是诱发肺癌的原因之一。     

肺癌组织中RKIP基因启动子区甲基化状态观察

目的观察肺癌组织中Raf激酶抑制蛋白（RKIP）基因启动子区甲基化状态变化,探讨其与肺癌临床病理特征的关系。方法采用RT-PCR和甲基化特异性PCR法（MSP）分析肺癌及相应癌旁肺组织中RKIP基因表达情况及其启动子区甲基化状态。结果肺癌组织中RKIP基因启动子区甲基化率为45．8％（27／56）,明显高于癌旁肺组织的13．3％（2／56）,P〈0．05。所有甲基化的肺癌组织中RKIP基因均无表达。有淋巴结转移的43例肺癌组织中,27例RKIP基因启动子甲基化;无淋巴结转移的40例中,11例RKIP基因启动子甲基化（P〈0．05）。结论肺癌组织中RKIP基因失表达与其启动子区甲基化有关,这可能是肺癌发生发展以及转移的原因之一。     

肺鳞癌组织中FHIT的表达变化及意义

采用免疫组化SP法检测36例肺鳞癌组织及11例癌旁组织中的脆性氨酸三联体（FHIT）。结果肺癌组织中FHIT蛋白阳性表达率为48．7％,显著低于癌旁组的90．9％（P〈0．01）。高、中分化肺癌组织中FHIT蛋白阳性表达率为60．O％,显著高于低分化组织的36．8％（P〈0．01）;有淋巴结转移的肺癌组织FHIT蛋白阳性表达率为43．7％,显著低于无淋巴转移者的75．0％（P〈0．05）;Ⅰ＋Ⅱ期肺癌组织中FHIT蛋白阳性表达率为52．6％,显著高于Ⅲ期的45．0％（P〈0．05）。认为FHIT在肺鳞癌的发生、发展中发挥作用,可作为判定肺鳞癌发生及转移能力的指标之一。     

端粒酶活性在原发性肝癌中的表达及临床意义

目的 探讨端粒酶活化在原发性肝癌发生发展中的作用。方法 采用端粒重复序列扩增技术（ＴＲＡＰ），对４２例手术切除的原发性肝细胞癌及癌旁组织的端粒酶活性进行检测。结果 ４２例肝癌组织有３４例检出端粒酶活性，阳性率为８１．０％。４２例癌旁组织有１２例检出端粒酶活性，阳性率为２８．６％。端粒酶活性与肿瘤大小、组织学分级及肿瘤转移无显著相关。结论 原发性肝癌组织端粒酶活性的检测对阐明原发性肝癌的发病机理，癌     

癌性胸水细胞端粒酶活性的检测意义

目的：研究癌性胸水细胞端粒酶活性的检测意义。方法：采用端粒重复序列扩增－核酸杂交分析法检测癌性胸水细胞端粒酶活性，并将检测结果与细胞学诊断进行比较。结果：49例诊断明确的癌性胸水细胞样本中23例细胞学阳性胸水有20例端粒酶阳性，5例细胞学可疑胸水有4例端粒酶阳性，21例细胞学阴性有14例端粒酶阳性。细胞学诊断和端粒酶检测总阳性率分别为46．9％（23／49）和77．6％（38／49），二联合检测率为83．7％（41／49）。结论：胸水细胞端粒酶活性的检测有助于癌性胸水的诊断，弥补胸水细胞学诊断的不足。     

Telomerase activity in human bladder tumors and bladder washing specimens

Telomerase appears to be an important factor for the control of cellular proliferation and tumorigenesis. Enzyme activity dramatically increases in almost all human tumors. The purpose of our study was to evaluate the role of telomerase activity as a marker for bladder cancer diagnosis and follow-up. By using the PCR-ELISA based on the TRAP (telomerase repeat amplification protocol) method, telomerase activity of bladder tumors (n = 77), normal-appearing adjacent tissues (n = 21) and bladder washings (n = 37) were analyzed. Telomerase activity was detected in 87% (67/77) of cancer tissues and in 38% (8/21) of normal-appearing adjacent tissues. However, the levels of enzyme activity were significantly higher in cancer tissues than in normal-appearing adjacent tissues (p < 0.05). Telomerase activity in bladder cancer tissues was not correlated to the tumor stage or grade. During a 26 months follow-up period, disease progression occurred in 66.7% of patients with invasive tumors where telomerase activity of the normal-appearing adjacent tissue was detectable, as compared to only 14.3% for patients who showed undetectable telomerase activity in adjacent, normal-appearing tissues (p = 0.094). When telomerase activity of bladder washing fluid was compared with its corresponding tumors, sensitivity of detection was 81% and specificity was 75%. In contrast, urine cytology only yielded a sensitivity of 31% in the detection of cancer. The detection ability between telomerase activity measurement in washing fluid and cytological examination had a trend toward the telomerase measurement identifying more cancer cases than the cytologic examination (p = 0.07). In conclusion, telomerase activity is present in early-stage bladder cancer and is a potential molecular marker for bladder tumors diagnosis. The expression of telomerase activity in normal-appearing mucosa adjacent to bladder tumor is probably an indicator of disease progression. Using the telomerase activity to detect exfoliated cells in bladder washing fluids could be a useful method in adjunct to urine cytology and cystoscopy in establishing the diagnosis and follow-up of bladder cancer.     

肺癌、良性实质性病变和正常肺组织端粒酶逆转录酶定位表达的研究

目的研究端粒酶在肺良、恶性病变和正常肺组织中表达的定位,以阐明端粒酶在上述疾病发病机制中的作用,以及在正常肺组织中的生物学意义。方法对38例肺癌、7例良性实质性病变及35例病灶旁正常肺组织标本用核酸原位杂交和免疫组织化学技术对端粒酶逆转录酶（hTERT）mRNA和蛋白质的定位表达进行了研究,同时与端粒酶活性进行了对比。结果肺癌、良性实质性病变和病灶旁正常肺组织端粒酶活性阳性分别为31/38、3/7、7/35（P<0.001）;肺癌组织癌细胞、肺良性实质性病变中的淋巴细胞、纤维母细胞、巨噬细胞等以及病灶旁正常肺组织小气道、细支气管上皮的部分细胞和少部分肺泡上皮细胞hTERT蛋白有阳性表达,在有淋巴小结的肺组织其生发中心可见hTERT表达,多数巨噬细胞也有hTERT阳性表达。hTERTmRNA所表达的细胞与免疫组织化学结果类似。肺癌hTERT免疫染色平均吸光度比较鳞癌A值比腺癌略高,Ⅲ期比Ⅰ期、Ⅱ期稍高,分化程度越低A值也越高,但P>0.05。结论(1)端粒酶除了参与肺癌的发病机制外,也参与肺良性实质性病变发生发展。（2）端粒酶对于维持正常肺组织支气管上皮细胞、肺泡巨噬细胞和淋巴细胞的功能可能有重要的生物学意义。(3)端粒酶是一种增殖指标,而非肺癌的恶性特异性标志,端粒酶作为肺癌诊断标记物时可能有一定的假阳性。     

Quantitative determination of telomerase activity in breast cancer and benign breast diseases.

Telomerase plays an important role in maintaining the stability of chromosomes. This ribonucleoprotein prevents chromosome ends (telomeres) from gradual loss with each cell division. It enables tumor cells to maintain telomere length, allowing indefinite replicative capacity. Telomerase activity has been detected in the majority of tumor and germ cells and in immortalized cell lines. Quantitative telomerase PCR-ELISA (TeloTAGGG Telomerase PCR ELISA(PLUS)) was evaluated for distinguishing benign and malignant breast tissue. Activity of telomerase was determined in 27 samples of fibrocystic and dysplastic tissues, 28 fibroadenomas and phylloid tumors, and 154 breast cancer tissues; 59 specimens were analyzed retrospectively. Analytical precision and linearity of the assay was tested using breast carcinoma cell line ZR-75-1 and breast tumor tissue extracts. About 4% of tumor samples were excluded from analysis due to interferences in the PCR reaction. Relative telomerase activity differed significantly in the groups of dysplastic tissues, fibroadenomas and carcinomas. The highest activity was found in breast cancer tissue. This method can identify breast cancer tissue with 73% clinical sensitivity and 93% specificity as compared to benign breast tumors. We did not find a correlation between telomerase activity and the tissue levels of estrogen and progesterone receptors, HER-2/neu oncoprotein concentration, tumor size, and lymph node positivity. Probability of disease-free survival was significantly lower for patients with telomerase activity higher than median value. As the assay for telomerase activity has very high analytical sensitivity and high specificity for cancer cells, this routinely used method may prove useful for distinguishing malignant phenotype of breast tissues.     

端粒酶逆转录酶和c-myc基因在肺癌中表达的研究

目的　研究端粒酶逆转录酶 (hTERT)和c myc基因在肺癌中的表达及其与端粒酶活性的关系。方法　采用端粒重复序列扩增 (TRAP)法对肺癌组织以及癌旁非癌肺组织的端粒酶活性进行测定。用原位逆转录聚合酶链反应 (RT PCR)和免疫组织化学技术分别检测上述组织标本中hTERTmRNA与c myc蛋白表达。结果　端粒酶活性在肺癌组织中阳性率为 73 .2 % (3 0 /4 1) ,在癌旁非癌肺组织为 6.3 % (2 /3 2 ) ,P <0 .0 1。肺癌及癌旁非癌肺组织中hTERTmRNA阳性表达率分别为 78.0 % (3 2 /4 1)和 9.4% (3 /3 2 ) ,P <0 .0 1,c myc蛋白阳性表达率分别为 80 .5 % (3 3 /4 1)和 15 .6% (5 /3 2 ) ,P <0 .0 1。肺癌组织hTERTmRNA表达与端粒酶活性呈显著正相关 (r =0 .70 7,P <0 .0 1) ,同时c myc蛋白与hTERTmRNA表达亦呈明显正相关 (r =0 .60 9,P <0 .0 1)。 结论　hTERTmRNA表达增强不仅参与肺癌的发生 ,而且可能是调节肺癌端粒酶活性的限速步骤。c myc蛋白可能通过激活hTERTmRNA表达而促进肺癌的形成     

端粒酶基因hTR在乳腺癌中的表达

用原位杂交的方法检测端粒酶基因hTR在46例乳腺及其癌旁组织，53例乳腺良性病变中的表达和分布情况。结果显示，46例乳腺癌中hTR表达阳性率为91．3％；癌旁组织中为0；53例乳腺良性病变中为3．9％。乳腺癌组织中hTR的表达与癌旁组织、良性病变比较差异有显著性（P均＜0．01）。乳腺癌组织中hTR的表达与肿瘤分期相关（P＜0．05），与病理类型、淋巴结转移、肿瘤大小无相关性（P均＞0．05）。认为端粒酶基因hTR表达检测在乳腺癌诊断中有一定价值。     

大肠癌及大肠息肉中端粒酶、PTEN的表达及意义

目的探讨端粒酶和PTEN在大肠癌及大肠息肉中的表达与病理特征的关系及临床意义。 方法应用免疫组织化学(S-P)法检测68例大肠息肉、25例息肉旁正常组织、15例结直肠癌癌灶组织、9例癌旁组织中端粒酶和PTEN蛋白的表达。 结果端粒酶在大肠癌组织中的阳性率(80.00%)显著高于癌旁组织(22.22%)、大肠息肉组织(19.12%)和息肉旁组织(0.00%)。PTEN在大肠癌组织中的阳性率(20.00%)显著低于癌旁组织(77.78%)、大肠息肉组织(54.41%)和正常大肠黏膜组织(80.00%)。在大肠癌及大肠息肉中,端粒酶和PTEN均成负相关(r＝－0.516;r＝－0.261)。端粒酶与息肉大小(r＝0.262)关系密切。端粒酶和PTEN的表达与大肠癌是否有淋巴结转移关系密切(r＝0.560;r＝－0.707)。 结论PTEN在大肠癌中的表达缺失,在腺瘤性息肉表现出缺失倾向,端粒酶和PTEN在大肠癌、大肠息肉中的表达呈负相关。两者的变化可能与结直肠肿瘤的恶性程度和疾病进展有关系。     

改进TRAP-PCR检测端粒酶活性对早期胃癌的诊断

目的探索端粒酶活性早期诊断恶性实体瘤的新方法．方法通过胃癌组织的分离和肿瘤细胞系的培养，用改良ＴＲＡＰＰＣＲ方法，来检测其端粒酶的活性．结果胃癌组织及肿瘤细胞系的端粒酶总表达率９３％（２５／２７），其中细胞系ＭＫＮ２８，ＭＫＮ４５，ＨＨＣＬ，ＳＧＣ７９０１及Ｔｃａ８１１３均为阳性表达；胃癌远隔的正常粘膜标本表达率为０％（０／２０）；其中有２００个肿瘤细胞存在便可测得其端粒酶的活性．结论胃癌组织及肿瘤细胞系的端粒酶表达，具有很强的肿瘤特异性和较高的表达率；改进ＴＲＡＰＰＣＲ方法具有提高恶性实体瘤早期诊断的潜在价值     

Real-time quantitative PCR of telomerase mRNA is useful for the differentiation of benign and malignant pancreatic disorders

The presence of telomerase activity has been proposed as a specific and sensitive marker for malignant tissue, and positivity rates of up to 95% have been reported in pancreatic cancer. In the present study telomerase activity analysis was reevaluated in 29 pancreatic cancer tissues compared with 36 chronic pancreatitis tissues and 21 normal controls, and a study was made of whether malignant and benign pancreatic disorders can be better differentiated using a novel technique real-time quantitative PCR analysis-analyzing telomerase mRNA expression. Telomerase activity was present in 35% (10 of 29) of pancreatic cancer samples, 3% (one of 36) of chronic pancreatitis samples, and none of the normal pancreatic tissue samples in the TRAP assay. Real-time quantitative PCR analysis revealed the presence of telomerase mRNA expression in 50% (10 of 20) of normal, 86% (31 of 36) of chronic pancreatitis, and 90% (26 of 29) of pancreatic cancer samples. However, quantification of the expression data revealed that the relative increase above normal was 5.5 (range, 3.5-8.6) for chronic pancreatitis and 23.9 (range, 18.6-30.7) for pancreatic cancer samples (p < 0.01). No relationship was found between telomerase activity and the fold increase of telomerase mRNA above normal and gender, patient age, tumor stage, or tumor grade. These data indicate that detection of telomerase activity using the TRAP assay has limitations in differentiating benign and malignant pancreatic disorders. However, telomerase mRNA analysis by real-time quantitative PCR analysis allows a highly sensitive detection and differentiation of pancreatic cancer from normal pancreas and chronic pancreatitis and thereby may serve as a new reliable, easy, and effective diagnostic tool for cancer diagnosis.