喉癌及癌旁组织端粒酶活性检测在喉癌手术切缘定界中的意义

目的探讨喉鳞状细胞癌、相应癌旁切缘组织的端粒酶活性表达及其在喉癌手术定界研究中的意义.方法采用多聚酶链反应-酶联免疫吸附法(PCR-ELISA)对47例喉鳞状细胞癌、21例不同距离的相应癌旁组织、20例喉炎性息肉组织的端粒酶活性进行定量检测.结果47例喉癌组织中39例端粒酶呈阳性(83%),而20例喉炎性息肉组织端粒酶均为阴性,两组具有显著性差异(P＜0.001).21例癌旁5 mm处的切缘组织中有6例端粒酶呈阳性(28.6%),但其活性水平均低于相应原发肿瘤组织.而10 mm处的切缘组织端粒酶均呈阴性,两组差异有显著性(P＜0.001).结论端粒酶在喉癌组织中有较高的表达,在不同距离的癌旁切缘组织中有不同程度的表达.喉癌及癌旁组织端粒酶活性可能作为预测喉癌浸润性的指标.可望从分子水平评估喉癌手术的切缘状态,以期为指导喉癌手术分子定界提供客观而有价值的资料.     

喉癌中Omi/HtrA2的表达及作用初探

目的:检测Omi/HtrA2在喉癌组织中的表达,初步探讨其在喉癌发生、发展中的意义。方法:采用RT-PCR与免疫组织化学方法,检测20例喉鳞状细胞癌及12例声带息肉手术切除标本中Omi/HtrA2mRNA与蛋白的表达情况。结果:癌旁正常组织中未检测到Omi/HtrA2蛋白表达,而在喉癌组织与声带息肉组织中均有表达。Omi/HtrA2基因在高分化喉鳞癌组织中的表达水平高于低分化型喉鳞癌组织、声带息肉以及癌旁的正常组织(P<0.01)。结论:Omi/HtrA2在喉鳞状细胞癌中有表达,表明Omi/HtrA2的促凋亡作用参与了喉癌的发生发展过程,为癌症治疗提供了新的理论基础。     

肝细胞癌端粒酶活性及hTERT mRNA表达与术后早期复发的关系

目的：研究肝细胞癌端粒酶活性及人端粒酶逆转录酶（hTERT）mRNA表达与肝细胞癌术后早期复发的关系。方法：采用ELISA—TRAP法检测60例肝癌组织及其癌旁组织端粒酶活性，RT—PCR法检测hTERT mRNA表达，5例正常肝脏组织作为对照。分析端粒酶活性及hTERT mRNA表达与临床病理之间的关系。结果：肝癌组织端粒酶活性及hTERT mRNA表达阳性率分别为86．7％（52／60）及90％（54／60），癌旁组织端粒酶活性及hTERT mRNA表达阳性率分别为40％（24／60）及43．3％（26／60）。正常肝脏组织均未检测到端粒酶活性及hTERT mRNA表达。癌旁组织端粒酶活性及hTERT mRNA表达与术后早期复发及包膜浸润、门静脉侵犯、肝内转移等恶性肿瘤的恶性生物学行为有关。结论：癌旁组织端粒酶活性及hTERT mRNA表达可能是肝细胞癌术后早期复发的预后指标。     

子宫颈病变中端粒酶活性的表达

目的：探讨宫颈糜烂，宫颈上皮内瘤样变（CIN）及宫颈癌中端粒酶的表达及其意义，方法：采用TRAP法检测30例宫颈糜烂，16例CIN组织，20例宫颈癌及20例正常宫颈组织中端粒酶活性。结果：宫颈糜烂者端粒酶活性阳性率为3．0％（1／30），CIN端粒酶活性阳性率为56％（9／16），20例宫颈癌组织中端粒酶阳性表达率为805（16／20），正常宫颈组织中端粒酶表达阴性。恶性肿瘤与正常或良性病变组织中端粒酶活性差异有显著性（P＜0．05）。结论：端粒酶的活化在宫颈癌的发生过程中起重要作用，端粒酶活性检测可能成为宫颈癌及宫颈癌前病变早期诊断诊断和病情监测的指标。     

食管癌端粒酶活性检测的意义

目的 探讨食管癌组织中端粒酶活性检测的临床意义。方法 采用聚合酶链反应－酶联免疫吸附法（ＰＣＲ－ＥＬＩＳＡ）检测了４３例食管癌组织及相应癌旁组织,８例食管良性疾病组织端粒酶活性表达。结果 ４３例食管癌组织中３６例端粒酶活性阳性表达,阳性率为８３．７％,而癌旁组织４例阳性,阳性率为９．３％,８例良性疾病食管组织端粒酶表达均为阴性。食管癌端粒酶检测阳性率明显高于癌旁组织及良性疾病食管组织。食管癌组织端     

口腔癌、癌前病变及癌前状态中抑癌基因PTEN的表达及其临床意义的研究

目的研究PTEN蛋白在口腔鳞状细胞癌、癌前病变及癌前状态组织中的表达及其与口腔鳞状细胞癌的临床病理及预后的关系。了解PTEN蛋白表达在口腔癌发生、发展中的作用。方法免疫组织化学法检测10例正常口腔粘膜、14例口腔扁平苔癣、20例口腔白斑、30例癌旁组织及46例口腔鳞状细胞癌组织中PTEN蛋白的表达水平,并进行半定量分析。结果正常口腔粘膜、口腔扁平苔癣、口腔白斑、癌旁组织及鳞状细胞癌组织中PTEN蛋白表达总阳性率分别为70．00％（7／10）、57．14％（8／14）、45．00％（9／20）、73．3％（22／30）及19．57％（9／46）。口腔白斑、扁平苔癣及鳞状细胞癌组织中VrEN蛋白表达水平显著低于正常口腔粘膜（P〈0．05）。口腔鳞状细胞癌组织中PTEN蛋白表达水平显著低于患者自身癌旁组织（P〈0．01）。结论PTEN蛋白表达水平降低或缺失在口腔鳞状细胞癌的发生发展过程中可能起重要作用。免疫组织化学法检测PTEN蛋白的表达对口腔癌早期诊断和预后判断的价值有待进一步研究。     

肝癌及癌旁组织中端粒酶检测的临床意义

目的研究端粒酶作为原发性肝细胞癌（ＨＣＣ）肿瘤标志物的可能性。方法采用ＴＲＡＰ方法检测了３３例原发性肝细胞癌及其３３例癌旁组织、４例肝转移癌及其４例癌旁组织、６例肝良性肿瘤和６例正常肝组织中的端粒酶活性。结果３３例原发性肝细胞癌组织中，有３０例端粒酶表达阳性，其阳性率为９０．９％。３３例癌旁组织中，有９例端粒酶表达阳性，其阳性率为２７．３％。４例肝转移癌端粒酶活性均阳性，４例癌旁组织中，２例端粒酶表达阳性。６例肝良性肿瘤中，仅１例端粒酶表达阳性。６例正常肝组织端粒酶表达均阴性。肝癌组织端粒酶表达与肿瘤临床病理特征无关。结论肝癌组织中普遍存在端粒酶活性表达，而良性和正常肝组织中端粒酶活性较少表达。端粒酶有可能成为诊断原发性肝细胞癌的肿瘤标志物     

LYVE-1与血管内皮生长因子-C在喉鳞状细胞癌中的表达和意义

目的通过测定喉鳞状细胞癌组织内LYVE-1和血管内皮生长因子-C（VEGF-C）的表达情况,为喉鳞状细胞癌转移和预后的判定以及治疗提供一定的理论依据。方法通过免疫病理学方法检测LYVE-1的表达并计数喉鳞状细胞癌组织中淋巴管密度（LVD）,RT-PCR法检测喉鳞状细胞癌中VEGF-C的表达,使用统计学方法对LVD和VEGF-C表达进行分析。结果喉癌组织内存在LYVE-1（＋）的管腔样结构,LYVE-1在喉癌组织和癌旁正常组织之间表达差异有统计学意义（P〈0．05）,喉癌组织中VEGF-C mRNA的平均水平与正常组织之间差异有统计学意义（P〈0．05）,瘤组织比正常组织高4-5倍,而且喉癌组织内VEGF-C表达与LVD之间存在相关性。结论喉鳞状细胞癌瘤组织中存在淋巴管;VEGF-C mRNA在喉鳞状细胞癌组织中的表达要比正常对照组织高,且VEGF-C mRNA的高表达可能会通过促进瘤内淋巴管的增生来促进喉癌的淋巴结转移。     

hOT7T175蛋白在喉鳞状细胞癌中的表达及临床意义

目的:检测hOT7T175蛋白在喉鳞状细胞癌中的表达并探讨其临床意义。方法:收集我院病理科喉鳞状细胞癌60例及癌旁喉组织（术后病理证实）20例。应用免疫组织化学方法（IHC）检测hOT7T175蛋白在喉鳞状细胞癌和癌旁喉组织中的表达情况。结果:喉鳞状细胞癌组织和癌旁喉组织中hOT7T175蛋白阳性率分别为58.33％和95.00％,二者差异有统计学意义（P<0.05）。hOT7T175蛋白与临床分期、病理分级、淋巴结转移分级密切相关。结论:hOT7T175蛋白在喉鳞状细胞癌中的表达率显著低于癌旁喉组织;hOT7T175基因可能与喉鳞状细胞癌的发生、发展过程有关。     

胃癌发生过程中端粒酶活性测定及临床意义

目的：通过检测胃癌及癌前病变组织中端粒酶活性，探讨其在胃癌发生发展中的作用及临床意义。方法：采用SP免疫组化法检测40例胃癌和18例癌前病变（其中慢性萎缩性胃炎伴肠化生10例，非典型增生8例）端粒酶活性。结果：胃癌及癌前病变中端粒酶阳性率分别为92．5％（37／40）和33．3％（6／18），两者比较差异有统计学意义，P〈0．05。端粒酶激活与胃癌的分化程度、病理分期和淋巴结转移无明显相关性，P〉0．05。结论：端粒酶活性表达与胃癌的发生密切相关，利用免疫组化检测端粒酶活性可作为胃癌早期诊断的参考指标，并应对阳性病例进行跟踪随访。     

胃癌及癌前病变组织中端粒酶RNA表达及其临床意义

目的 探讨胃癌及癌前病变胃粘膜端粒酶RNA与端粒酶活性检测及临床意义。方法 分别采用原位逆转录－PCR、端粒重复序列扩增法（TRAP）检测114例胃粘膜组织标本端粒酶RNA与端粒酶活性,其中包括慢性浅表性胃炎（CSG）32例、不典型增生（AH）34例、胃癌（GC）48例。结果 原位逆转录－PCR检测胃粘膜活检标本的端粒酶RNA阳性率为57．9％（66／114）,显著高于TRAP法检测端粒酶活性检出率（44．7％,51／114,P＜0．05）。在不同胃粘膜病变中,AH及GC组的端粒酶RNA阳性率分别为52．9％、100％,而CSG组胃粘膜中未检出端粒酶RNA;AH及GC组的胃粘膜端粒酶RNA阳性率分别显著高于CSG（P＜0．05）,而GC组端粒酶RNA阳性率亦明显高于AH组（P＜0．05）。端粒酶RNA主要分布于胃粘膜癌细胞及癌前病变上皮细胞的胞核内。结论 端粒酶RNA表达与胃癌的发生密切相关。原位逆转录－PCR检测胃粘膜端粒酶RNA可能是较端粒酶活性更灵敏的生物学指标,对胃粘膜癌变的预测和早期诊断有重要价值。     

不同病变胃粘膜端粒酶活性及其亚单位的检测

目的：探讨端粒酶及其亚单位（ＴＰ１、ｈＴＲ、ｈＴＲＴ）在胃粘膜癌变过程中的作用。方法：端粒酶的检测采用端粒末端重复序列扩增技术（ｔｅｌｏｍｅｒｉｃ　ｒｅｐｅａｔ　ａｍｐｌｉｆｉｃａｔｉｏｎ　ｐｒｏｔｏｃｏｌ,ＴＲＡＰ）,端粒酶亚单位的检测采用ＲＴ－ＰＣＲ法。结果：端粒酶阳性检出率在慢性萎缩性胃炎、肠上皮化生、异型增生及胃癌中分别为２４．６％（１４／５７）、３８．９％（７／１８）、３７．５％（３／８）及９２．３％（６０／６５）,正常胃粘膜未检测到端粒酶活性,与以上各病变组织有显著性差异（Ｐ＜０．０５～０．０１）,而慢性萎缩性胃炎、肠上皮化生和异型增生组织中端粒酶阳性率亦明显低于胃癌组织（Ｐ＜０．０１）;端粒酶的表达与临床病理指标无相关性;ＲＴ－ＰＣＲ定性检测发现端粒酶亚单位ｈＴＲ和ＴＰ１在大多数胃粘膜中都有表达,而ｈＴＲＴ主要在胃癌及部分癌前组织中表达,且在胃癌中ｈＴＲＴ的表达与端粒酶活性之间具有明显的相关性（Ｐ＜０．０１）。结论：端粒酶不仅在胃癌组织中高表达,在部分癌前组织中也有表达。提示端粒酶在胃癌的发生过程中可能具有重要作用;端粒酶亚单位ｈＴＲＴ不仅可能作为胃癌的诊断指标,而且还可能作为胃癌基因治疗的靶     

48. The value of CT scan and detection of telomerase activity in biopsy specimens for early diagnosis of lung carcinoma

Objective: To evaluate the diagnostic value of telomerase activity in the specimens of biopsy with bronchoscopy or cutting needle. Methods: Telomerase activity was measured in the biopsy apecimens taken from 52 patients suspected of having early lung cancer by CT scan. The PCR based silver staining telomeric repeat amplification protocol (TRAP) was used for detection of telomerase activity in 22 patients with early lung cancer (T1N0M0). Control study was made on the specimens taken from 24 patients with benign disease (cyst 3, TB 6, pseudtumor 5, pneumonjia 10). Results: The positive rates of telomerase activity were 86.45% (19/22) and 4.2% (1/24) in early lung cancers and benign lesions respectively (P<0.01). It was significantly higher in early lung cancers than in benign disease. All cases were diagnosed with surgical pathology and following for 2 years. Conclusion: Detecting telomerase activity in preoperative bronchoscope and cutting needle biopsy specimens may contribute to diagnosis of early lung cancer.     

乳腺癌组织中端粒酶活性的定量检测及其与临床病理的关系

背景与目的:端粒酶是一种在细胞永生化及癌症发生过程中起重要作用的核蛋白酶。最近关于乳腺癌中端粒酶活性与预后因素间的关系,因研究方法的差异而呈现出不一致的报道。本研究旨在建立一种可行的银染端粒酶定量检测法,以探讨乳腺癌中端粒酶活性与临床病理学预后因素间的关系。方法:运用银染端粒重复序列扩增法(SS-TRAP)检测了52例新鲜乳腺癌及其癌旁组织,32例乳腺良性病变和14例正常乳腺组织中端粒酶的活性表达,对结果予以定量并结合临床资料进行分析。结果:乳腺癌、癌旁组织、良性病变及正常乳腺组织中端粒酶阳性率分别为90.38%(47/52)、28.85%(15/52)、31.25%(10/32)、0(0/14)。端粒酶分别为36.91±15.35、8.27±4.37、14.10±5.28、0(TPG单位),单因素方差分析结果显示,乳腺癌组端粒酶活性水平明显高于其他3组(P值均<0.01)。Logistic回归分析结果表明,乳腺癌中端粒酶的表达与病理类型、分化程度明显相关(P=0.003及0.004),即随着肿瘤的进展,端粒酶活性亦增加。其中,浸润性非特殊癌端粒酶活性水平明显高于浸润性特殊癌(P<0.05);中、低分化癌端粒酶活性均高于高分化癌(P<0.05),中、低分化癌间无显著性差异(P>0.05)。端粒酶活性表达在患者病程、年龄、绝经状况间均无显著性差异(P>0.05)。结论:与经典TRAP     

Evaluation of telomerase activity in bronchial lavage as a potential diagnostic marker for malignant lung disease

Telomerase is a ribonucleoprotein DNA polymerase that maintains the telomeric region of chromosomes lost during successive rounds of cell division. We used the telomeric repeat amplification protocol (TRAP) assay to examine telomerase activity in bronchial lavage (BL) samples from individuals undergoing diagnosis of lung cancer. Telomerase activity was detected in 17 (47%) of 36 samples examined. In particular, 16 (70%) of 23 BL specimens obtained from lung cancer patients showed detectable telomerase activity, while only 1 of 13 (8%) specimens obtained from patients without lung cancer demonstrated activity (P=0.00038). Moreover, 9 (90%) of 10 BL specimens, which were cytologically positive for lung cancer, were also positive for telomerase activity, while 7 (54%) of 13 cytologically negative BL specimens for lung cancer showed detectable telomerase activity. Detection of telomerase activity combined with cytology were able to identify 17 (74%) of 23 lung cancer cases whereas cytology alone identified 10 (43%) of 23 such cases (P=0.035). Our findings indicate that telomerase is a specific marker for malignant lung disease and a potential complementary tool to cytology in the diagnosis of certain lung cancer cases.     

Telomerase activity: comparison between fine-needle aspiration and biopsy specimens for the detection of tumor cells.

BACKGROUND: The purpose of the current study was to determine telomerase activity as a sensitive biomarker for the detection of malignant cells in fine-needle aspiration (FNA) specimens. METHODS: FNA specimens with parallel samples of fresh tumor tissue were obtained from surgical specimens after surgical excision. Using a polymerase chain reaction-based telomeric repeat amplification protocol (TRAP) assay, telomerase activity was determined systematically in FNA specimens (n = 21) and corresponding available tissue biopsy specimens (n = 16) containing malignant cells. In addition to a case of myelolipoma, normal counterparts for 3 of 16 cancer cases, including both biopsy and FNA specimens, also were available for the determination of telomerase activity. RESULTS: Telomerase activity was observed in 14 of 16 of the FNA specimens (88%) and 15 of 16 of the corresponding biopsy specimens (94%). Telomerase activity was detected in both the biopsy specimen and the corresponding FNA specimen, with one exception (a case of mucinous adenocarcinoma of the cecum). In contrast, specimens from three normal tissue biopsies and FNA specimens of normal tissue adjacent to the malignant lesions, as well as the myelolipoma, exhibited no telomerase activity. It is interesting to note that both tissue biopsy specimens and FNA specimens from a patient with high grade sarcoma were negative for telomerase activity. The examination of hematoxylin and eosin-stained adjacent tissue biopsy sections or FNA smears revealed similar low populations of lymphocytes, including those cases that were negative for telomerase activity. There was agreement in the detection of telomerase activity between tissue biopsies and their corresponding FNA specimens in 15 of the 16 patients, indicating a 94% concordance rate (95% confidence interval, 70%, 98%). CONCLUSIONS: The results of the current study clearly suggest that the telomerase activity in FNA specimens was comparable to that of their corresponding biopsy specimens, and that this activity was associated with the presence of malignant cells. The TRAP assay has potential for use in the detection of malignant cells in FNA specimens, particularly cases in which the cytology is not characteristically malignant and/or is present in insufficient numbers. Cancer (Cancer Cytopathol) Copyright 1999 American Cancer Society.     

Clinical significance of telomerase activation and telomeric restriction fragment (TRF) in cervical cancer

Telomerase activation was examined in 50 cases of cervical cancer, 27 normal cervix and five cervical cancer cell lines using the sensitive polymerase chain reaction (PCR)-based TRAP (telomeric repeat amplification protocol) assay. Telomeric restriction fragment (TRF) length of these specimens was measured by Southern hybridisation. Telomerase activation was common in cervical cancers and was detected in 46/50 cases (92%). Telomerase activity was weak in normal cervix and was detected only in 2/27 cases (7.4%). Telomerase activity was detected in all stages of cervical cancer suggesting that it is an early event in cancer progression. The clinical significance of telomerase activation was analysed in 47 squamous cell carcinoma of the cervix. High telomerase activity was more frequently detected in advanced diseases (100% in stage III and stage IV cervical cancers combined) compared with early diseases (68.6% in stage I and stage II cancers combined). The difference was statistically significant (P < 0.02). Telomerase activity was not statistically correlated with other clinical parameters examined. This is the first report of telomeric length in human cervical cancer. Both shortening and elongation of TRF length in cervical cancers was observed. Advanced cervical cancers tended to have a wider range of variation of TRF length compared with early disease and normal cervix. There was no obvious relationship between TRF length and the clinical parameters examined including clinical staging, differentiation status of tumour, human papilloma virus (HPV) infection, recurrence rate, tumour size and invasion depth. The clinical significance of TRF length appears to be limited in cervical cancers. Our results indicate that telomerase activity is closely associated with tumour cells and may be useful as a marker for detection of tumour cells in cervical biopsies.