胃癌组织中端粒酶活性与细胞凋亡的关系

目的为了探讨胃癌组织中端粒酶与细胞凋亡的关系。方法采用原位杂交和原位末端DNA片段标记技术对60例胃癌组织中端粒酶和细胞凋亡指数(AI)进行检测。结果60例胃癌中49例端粒酶阳性(81.7%)，55例(91.7%)可观察到凋亡细胞。32例淋巴结转移癌中端粒酶阳性率为93.8%，明显高于无淋巴结转移组的67.9%(P<0.05)；淋巴结转移组的AI为5.51±2.70，明显低于无淋巴结转移组的8.82±3.17(P<0.05)；淋巴结转移组胃癌中的端粒酶活性与AI呈负相关(P<0.05)。结论端粒酶活性和细胞凋亡可作为胃癌的生物学行为和预后的观察指标。     

胆管癌组织及胆汁中端粒酶活性的检测及意义

目的：检测胆管癌组织和胆汁中的端粒酶活性，以研究与端粒酶与胆管癌的关系及对胆管癌的诊断意义。方法：用PCR—ELISA方法检测20例胆管癌的癌组织和胆汁中端粒酶活性，同时用相同的方法检测20例正常胆管组织和胆汁中的端粒酶活性以作对照。结果：20例胆管癌组织中，端粒酶活性阳性为80％（16／20），胆汁中端粒酶活性阳性率75％（15／20）。正常胆管组织端粒酶活性阳性表达率为0（0／20），胆汁中端粒酶活性阳性率为0（0／20）。结论：端粒酶可能参与了胆管癌的发生发展过程，检测胆汁中端粒酶活性可有助于胆管癌的诊断。     

前列腺衰老逃逸现象的实验研究

目的:了解良性前列腺增生腺体中不同组织的端粒酶阳性细胞分布情况,从端粒酶角度研究前列腺增生的机制。方法:采用TRAPHybKit测定端粒酶活性。实验分两步。第一步,测定正常前列腺与良性增生腺体的端粒酶阳性率。第二步,测定前列腺增生腺体中增生结节和包膜的端粒酶活性。结果:13例正常前列腺端粒酶阳性率为15.38%(2/13),35例前列腺增生组织端粒酶阳性率为25.71%(9/35)。增生结节、包膜各33管中端粒酶阳性率分别为42.42%(14/33)和3.03%(1/33);前列腺增生组织中端粒酶阳性表达率显著高于正常前列腺组织,P<0.01,增生结节阳性率高于包膜组织,P<0.05。结论:前列腺增生组织中端粒酶活性升高,前列腺增生组织中端粒酶分布不均匀,增生结节含端粒酶阳性细胞高于包膜组织。提示前列腺的衰老逃逸可能与端粒酶活性有关。     

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     

前列腺衰老逃逸现象的实验研究

目的：了解良性前列腺增生腺体中不同组织的端粒酶阳性细胞分布情况，从端粒酶角度研究前列腺增生的机制。方法：采用TRAPHyb Kit测定端粒酶活性。实验分两步。第一步，测定正常前列腺与良性增生腺体的端粒酶阳性率。第二步，测定前列腺增生腺体中增生结节和包膜的端粒酶活性。结果：13例正常前列腺端粒酶阳性率为15．38％（2／13），35例前列腺增生组织端粒酶阳性率为25．71％（9／35）。增生结节、包膜各33管中端粒酶阳性率分别为42．42％（14／33）和3．03％（1／33）；前列腺增生组织中端粒酶阳性表达率显著高于正常前列腺组织，P〈0．01，增生结节阳性率高于包膜组织，P〈0．05。结论：前列腺增生组织中端粒酶活性升高，前列腺增生组织中端粒酶分布不均匀，增生结节含端粒酶阳性细胞高于包膜组织。提示前列腺的衰老逃逸可能与端粒酶活性有关。     

肝癌细胞中端粒酶活性的检测

目的　本实验为检测肝癌组织中是否有端粒酶活性，端粒酶活性与肝癌临床检测指标ＡＦＰ及高低转移潜能是否相关而设计。　方法　本实验用ＴＲＡＰ法从组织中抽提端粒酶，经反转录并ＰＣＲ 扩增后用地高辛标记的探针作斑点印迹杂交（ 肝癌标本部分） 和ＰＡＧ电泳加银染法显色（ 人肝癌高低转移裸鼠模型） 。　结果　本实验检测了１１ 例肝癌患者有肝癌组织和癌周组织的端粒酶活性，结果肝癌组织中端粒酶活性为９３－８８（单位：ＩＯＤ值） ，癌周组织端粒酶活性为２４－０９，两者的差别有统计学意义（Ｐ＜ ０－００１） 。正常组织（ｎ ＝ ５） 中未检测到端粒酶活性。端粒酶活性与ＡＦＰ浓度的相关系数ｒ＝０ ．７２，与肿瘤大小的相关系数ｒ＝ －０－２９８。人肝癌高转移模型裸鼠ＬＣＩＤ２０ 的端粒酶活性为（７７－６７±６－９３），低转移模型裸鼠ＬＣＩＤ３５ 的为８２－１６ ±１５－９３），两者之间并无显著性差异（ Ｐ＞ ０－０５） 。　结论　肝癌组织的端粒酶活性明显地高于癌周组织，正常组织中未检测到端粒酶活性。端粒酶活性与ＡＦＰ及肿瘤大小无关。端粒酶活性和肝癌的转移潜能可能不直接相关     

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     

胃癌端粒酶活性与DNA倍体的关系及临床意义

目的：探讨胃癌组织中端粒酶活性与DNA倍体的关系及临床意义。方法：用端粒酶重复序列扩增法（TRAP）检测胃癌组织中的端粒酶活性,用DNA图像分析仪（ICM）检测肿瘤细胞核DNA倍体。结果：胃癌组织中端粒酶的阳性表达率为81．25％,早期胃癌病例的阳性率为55．5％,进展期胃癌的阳性率87．2％。48例中异倍体肿瘤33例,占68．7％。33例异倍体胃癌中91％（30／33）显示端粒酶活性。结论：胃癌组织中端粒酶活性与DNA异倍体率有明显的相关性,端粒酶的表达与胃癌的病程进展有关。     

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。     

膀胱癌组织中端粒酶活性的检测及其意义

目的 研究端粒酶与膀胱移行细胞癌（TCC）生物学行为的关系。方法 方法 采用端粒酶重复序列扩增法及酶联免疫吸附性检测58例TCC组织、10例癌旁膀胱粘膜及10例正常膀胱粘膜组织的端粒酶活性。结果 TCC组织端粒酶阳性率为86．2（50／58）,不同病理分级、临床分期之间无显著性差异（P＞0．05）;癌旁组织中只有1例端粒酶阳性;正常组织中端粒酶均为阴性。结论 TCC组织中端粒酶活性阳性率明显高于正常膀胱粘膜组织及癌旁膀胱粘膜组织。各病理分级和临床分期之间端粒酶活性虽然无显著性差异（P＞0．05）,但端粒酶活性的强弱分布不同。端粒酶活性可作为TCC早期诊断、鉴别诊断及预测复发的肿瘤标志物之一。     

Significance of telomerase activity in the diagnosis of small differentiated hepatocellular carcinoma

Precise diagnosis of well-differentiated hepatocellular carcinoma (HCC) is sometimes difficult to establish. Telomerase activity was examined by telomeric-repeat-amplification protocol (TRAP) in 37 HCC nodules smaller than 3 cm in diameter, including 24 fine-needle-aspiration biopsy specimens, 22 non-tumor chronic-liver-disease tissues (9 chronic hepatitis and 13 liver cirrhosis) and 3 normal liver tissues. Telomerase activity was assayed by serially diluted samples and quantitated by using an internal telomerase assay standard (ITAS). Telomerase activity was detected in all HCC and in 11 of 22 non-tumor chronic-liver-disease tissues. Normal liver samples had undetectable telomerase activity. Cut-off level of telomerase activity for its practical usage in HCC diagnosis was tentatively set for 0.6 μg liver protein/assay at 10-cell equivalent activity of a gastric-cancer cell line, MKN-1. This level was twice the highest activity in non-tumor chronic liver disease therefore, telomerase activity in all non-tumor liver samples was below this level. The telomerase-positive incidence exceeding this cut-off level was 73% (11/15) in well-differentiated HCC, 94% (16/17) in moderately differentiated HCC and 100% (5/5) in poorly differentiated HCC. Well-differentiated HCC showed low positivity by other diagnostic markers. 21% by AFP, 0% by PIVKA-II and 13% by angiography. The detection of telomerase activity may thus be a useful additional tool for precise and early diagnosis of small differentiated HCC, even when diagnosis is inconclusive by conventional techniques. Int. J. Cancer 74:141-147, 1997. © 1997 Wiley-Liss, Inc.     

Surgical Significance of Telomerase Activity in Noncancerous Liver Tissue from Patients with Hepatocellular Carcinoma

Telomerase activity has been detected in tissue from noncancerous liver of patients with chronic liver disease, but its functional significance remains to be elucidated. We therefore evaluated the telomerase activity in surgically obtained noncancerous liver tissue from 20 hepatocellular carcinoma (HCC) patients. Two samples of noncancerous liver tissue were obtained from each patient: one from the parenchyma adjacent to the HCC nodules of the resected specimen; the other from the parenchyma distant from the HCC nodules of the remnant liver. Telomerase activity was assayed by a non-radioisotope quantitative system based on "TRAP-eze.'Five samples from the noncancerous liver tissue adjacent to the HCC nodules (25.0%) were telomerase-positive; all such cases showed high-grade malignant potential, such as intrahepatic metastasis and/or portal vascular invasion and infiltration of the fibrous capsule in the corresponding HCC nodules, and telomerase positivity showed neither a relationship with the histological activity index scores nor a correlation with liver function. Interestingly, no telomerase activity was detected in any of the 20 samples obtained from the parenchyma of the remnant liver. These results indicate that telomerase in noncancerous liver tissue is associated not with the hepatic condition accompanying HCC, but with the biological characteristics of the tumor itself, and may derive from infiltrating cancer cells. Determination of telomerase status may aid in designing more effective surgical procedures.     

原发性肝癌细针活检端粒酶活性检测的临床意义

目的:探讨原发性肝癌(HCC)经皮肝穿刺细针活检端粒酶活性检测的临床意义.方法:25例直径1.2～3cm的HCC进行了无水酒精注射治疗(PEIT).PEIT治疗前及PEIT治疗后1周在B超引导下经皮肝穿刺细针活检,所得微量肝癌组织采用非放射性同位素银染端粒重复序列扩增法(TRAP)行端粒酶活性检测及病理学检查.3例正常肝脏组织及5例肝硬变组织作为对照研究.结果:PEIT治疗前,病理学检查诊断率为80%(20/25),端粒酶活性阳性率为84%(21/25);PEIT治疗后1周,病理学检查未发现肿瘤细胞,3例端粒酶活性阳性(12%),其中1例经手术治疗后临床治愈,2例未手术治疗者1年内复发.其余22例端粒酶转阴的患者1年内未复发.3例正常肝脏组织及5例肝硬变组织端粒酶活性检测均阴性.结论:经皮肝穿刺活检微量肝脏组织端粒酶活性检测可应用于小肝癌的辅助诊断,在原发性肝癌PEIT治疗的疗效评价及预后判断方面具有一定的应用价值.     

Hop bitter acids inhibit tumorigenicity of hepatocellular carcinoma cells in vitro

Bitter acids (BAs) from the hop plant Humulus lupulus L. exhibit multiple beneficial biological properties with promising effects in cancer therapy and prevention, but information regarding the effects on hepatocellular carcinoma (HCC) is missing. Here, we used two different hop bitter acid extracts enriched for either α-acids or β-acids to obtain insight into whether biological activity varies between these two groups of BAs. At a concentration of 25 μg/ml, only the β-acid rich started to induce aspartate transaminase (AST) release, and a significant increase was detected with 50 μg/ml of both extracts. Already at lower concentrations both extracts led to a dose-dependent inhibition of proliferation, and migration was suppressed at a concentration as low as 5 μg/ml in HCC cells. The focus on different signaling pathways revealed an inhibition of ERK1/2 phosphorylation, downregulation of AP-1 activity and an alleviation of nuclear factor κB (NFκB) activity in HepG2 cells incubated with 5 μg/ml of both extracts, whereby the β-acid rich extract showed more pronounced effects. In conclusion, we identified ERK1/2, AP-1 and NFκB, which are important factors in tumor development and progression, as targets of hop BAs. Thus, these data suggest the potential use of BAs as functional nutrients for both prevention and treatment of HCC.