大肠肿瘤APC基因产物的检测

应用免疫组化方法检测大肠癌、腺癌、移行粘膜、正常粘膜、的ＡＰＣ基因蛋白产物Ｐ＾３００ＡＰＣ阳性率分别为２０％、６６．７％、６６．７、７０％，差别有显著意义，１５例大肠腺瘤中，４例伴重度非典型增生者Ｐ＾３００ＡＰＣ染色呈阴性（Ｐ〈０．０１）。表明：Ｐ＾３００ＡＰＣ的低表达是大肠肿瘤发生中的重要早期事件。     

大肠肿瘤APC基因产物的检测

应用免疫组化方法检测大肠癌、腺瘤、移行粘膜、正常粘膜、的ＡＰＣ基因蛋白产物Ｐ￣（３００ＡＰＣ）阳性率分别为２０％、６６．７％、６６．７、７０％，差别有显著意义。１５例大肠腺瘤中，４例伴重度非典型增生者Ｐ￣（３００ＡＰＣ）染色呈阴性（Ｐ＜０．０１）。表明：Ｐ￣（３００ＡＰＣ）的低表达是大肠肿瘤发生中的重要早期事件。     

大肠癌组织中多胺水平测定的意义

用反相高压液相色谱法检测４３例大肠癌组织及癌旁粘膜中的多胺（精脒，精胺）的水平，结果发现：癌组织中精脒含量明显高于正常大肠粘膜（Ｐ〈０．０５）。精脒／精胺比值明显高于正常大肠粘膜及癌旁粘膜（Ｐ〈０．００１）；低分化癌高于高分化癌（Ｐ〈０．０５）；ＤｕｋｅｓＢ期及Ｃ期癌组织中精脒含量及精脒／精胺比值均明显高于Ａ期（Ｐ〈０．０５）。表明精脒参与了大肠癌细胞的增殖和分化，精脒水平及精脒／精胺比值反映了大     

HSP70mRNA在人大肠癌中的高表达及其临床意义

目的 探讨热休克蛋白７０（ＨＳＰ７０）在人大肠癌中的表达情况。方法 用ＲＮＡ斑点杂交方法测定３２例大肠癌病人标本中ＨＳＰ７０ｍＲＮＡ的表达情况。结果 在人大肠癌组织中ＨＳＰ７０ｍＲＮＡ的表达增高，是正常大肠粘膜的１．９２倍（Ｐ〈０．００１），肿瘤旁大肠粘膜的１．６０倍（Ｐ〈０．００１），肿瘤旁组织又是正常组织的１．３１倍（Ｐ〈０．０１），结论 ＨＳＰ７０基因在人大肠癌中的表达增高。     

直肠癌与癌旁粘膜肿瘤相关抗原表达的研究

应用免疫组化方法检测２０例直肠癌患者癌组织及癌旁粘膜肿瘤相关抗原的表达以及癌旁粘膜粘液性质的变化。结果显示：Ｔ抗原在癌组织的表达明显高于癌近侧端１０ｃｍ处粘膜的表达，但与癌远侧端５ｃｍ处粘膜的Ｔ抗原表达则无明显差异。在距直肠癌远端５ｃｍ及近端１０ｃｍ处粘膜唾液酸粘蛋白反应分别为４５％（９／２０）和２０％（４／２０），癌旁远侧端５ｃｍ处粘膜Ｔ抗原与唾液酸粘蛋白均呈阳性反应者为４０％（８／２０），均呈阴性者为３０％（６／２０），两者之间差异显著（Ｐ＜０．０５）。但在癌旁近侧端１０ｃｍ处两者之间则无显著差异（Ｐ＞０．０５）。由此表明，直肠癌旁粘膜肿瘤相关抗原的异常表达及其分泌的粘液性质的改变可能提示癌旁粘膜存在某种程度的恶性潜势。     

大肠癌细胞端粒酶活性及端粒动力学研究

目的 探讨端粒动力学改变与端粒酶活化间的关系及其在大肠癌形成演化过程中的作用。方法 采用ＴＲＡＰ－ＥＬＩＳＡ法检测５８例大肠癌及其中３０例相应癌旁组织、远癌切端肉眼正常粘膜组织和６例良性大肠疾患端粒酶活性。Ｓｏｕｔｈｅｒｎ－ｂｌｏｔ－ＥＣＬ法对上述３０例大肠癌患者Ｔ、Ｐ、Ｎ不同位点和６例良性大肠疾患的粘膜进行平均端粒长度测定结果 Ｔ组ＴＲＦＳ值较Ｐ、Ｎ组明显缩短,而端粒酶活性表达Ｔ组（９１．４）,     

大肠癌及癌旁粘膜中p21,p185,PCNA的表达及意义

目的 探讨癌基因相关蛋白Ｐ２１,Ｐ１８５及增殖细胞核抗原（ＰＣＮＡ）在大肠癌癌旁移行粘膜、近端断端大肠粘膜、腺瘤型息肉中表达的意义及其与大肠癌的关系。方法 应用化学方法对４０例原发性大肠癌及癌旁移行粘膜,２０例腺瘤 肉中Ｐ２１,Ｐ１８５及ＰＣＮＡ的表达进行检测。结果 Ｐ２１,Ｐ１７８５及ＯＰＣＮＡ在大肠癌癌旁移行粘膜中的表达率分别为２２．５％＊（９／４０）,１７．５％（６／４０）,１００％（４０／     

大肠癌组织胃泌素与c—myc,c—fos表达的关系

目的 研究大肠癌组织中胃泌素与癌基因ｃ－ｍｙｃ、ｃ－ｆｏｓ表达的关系,以探讨胃泌素对大肠癌的促增殖作用机理。方法 采用免疫组化ＳＰ法检测４８例大肠癌患者癌组织及癌旁粘膜胃泌素和癌基因ｃ－ｍｙｃ、ｃ－ｆｏｓ的表达。结果 大肠癌组织胃泌素阳性率为３９．５８％,高分化腺癌明显高于低分化和粘液腺癌（Ｐ〈０．０５）。癌组织的ｃ－ｍｙｃ和ｃ－ｆｏｓ表达阳性率显著高于癌旁粘膜和正常粘膜。胃泌素阳性组癌组织ｃ－ｍｙｃ和ｃ－ｆｏｓ表达阳性率分别为７８．９４％和７３．６８％,胃泌素阴性组分别为３７．９３％和３１．０４％,差异均有显著性意义（Ｐ〈０．０５,Ｐ〈０．０１）。结论 部分大肠癌细胞通过自分泌方式产生胃泌素,可能通过增加ｃ－ｍｙｃ、ｃ－ｆｏｓ等癌基因的表达,从而刺激癌细胞的价值。     

大肠癌患者血清及癌组织中生长抑素水平测定的临床意义

检测４３例大肠癌患者血清、癌组织及癌旁粘膜中生长抑素水平。结果大肠癌患者术前空腹血清生长抑素含量明显低于对照组（Ｐ〈０．００１），根治术后明显高于术前（Ｐ〈０．０１），但仍明显低于对照组（Ｐ〈０．００１）。大肠癌组织中生长抑素含量明显低于正常大肠粘膜（Ｐ〈０．００１），亦低于癌旁粘膜（Ｐ〈０．００１），但血清及癌组织中生长抑素水平与大肠癌组织学类型及Ｄｕｋｅ＇ｓ分期均无显著关系。提示生长抑素对大肠     

大肠癌发生过程中p16蛋白的表达及其临床意义

目的：探讨Ｐ１６蛋白在大肠癌中的表达及意义。方法：采用免疫组化ＡＢＣ法观察８０例大肠癌、大肠腺瘤及正常粘膜中Ｐ１６蛋白的表达情况。结果：大肠癌组织Ｐ１６蛋是性表达率为４０．０％,低于正常粘膜及腺瘤中的表达（Ｐ〈０．０５）,其阳笥表达率在癌组织的分化程度、有无淋巴结转移及病人生存时间等方面有显著性差异（Ｐ〈０．０５）。结论：大肠癌组织中存在Ｐ１６蛋白的缺失和突变,且Ｐ１６蛋白在大肠癌癌细胞的增殖和分     

大肠癌端粒酶各组分表达及其与端粒酶活性的关系

目的：研究端粒酶的3种组分与其活性的关系,探讨端粒酶激活的关键因素。方法：采用TRAP法检测大肠癌组织及相邻正常黏膜组织中端粒酶活性,用RT－PCR检测端粒酶3种组分的表达。结果：在64例（85．33％）癌组织中检测到端粒酶活性,正常黏膜中没有端粒酶活性,两者差异有显著性（P＜0．01）;hTR和TP1基因的表达在癌组织和正常黏膜没有差别,hTERT在癌组织中的表达要明显高于正常黏膜,差别具有显著性（P＜0．01）,hTERT基因表达强度与端粒酶活性密切相关。结论：大肠癌端粒酶活性表达具有肿瘤特异性,hTERT在癌组织中的表达明显高于正常粘膜,大肠癌中端粒酶活性和hTERT基因表达强度密切相关,hTERT基因表达可能是端粒酶激活的关键因素。     

大肠癌患者粪便标本的端粒酶活性研究

目的 探讨通过粪便途径筛查大肠癌的可行性和新方法。方法 应用聚合酶链端粒重复扩增（PCR－TRAP）银染技术,研究了43例大肠癌患者粪便中脱落细胞的端粒酶活性表达。结果 62．8％的大肠癌患者粪便标本中有端粒酶阳性表达。患者粪便标本中端粒酶阳性表达与其大肠癌Dukes分期、淋巴结转移和癌肿部位未见显著相关（P＞0．05）。1例结肠腺瘤患者粪便标本端粒酶表达阳性,其腺瘤组织也存在端粒酶活性表达。粪便端粒酶检测的敏感性、特异性和阳性预测值分别为62．8％、95．7％和96．4％。结论 PCR－TRAP银染检测大肠癌患者粪便脱落细胞的端粒酶活性表达为改善大肠癌筛查方法和大肠癌诊断作了新的尝试。     

大肠癌组织端粒酶活性的检测及意义

探讨端粒酶活化在大肠癌发生发展中的作用。方法:采用PCR技术对32例手术切除大肠癌及其癌旁组织及4例大肠腺瘤的端粒酶活性进行检测。结果:32例大肠癌组织有25例检出端粒酶活性,阳性率为78.1％。2例绒毛状腺瘤检出端粒酶活性,2例管状腺瘤为阴性。端粒酶活化与肿瘤部位、大小、组织学分级、浸润深度、淋巴结转移及Dukes分期无显著相关。结论:检测大肠癌组织端粒酶活性对阐明大肠癌的发病机理、癌变危险性的预测及早期诊断均可能有重要意义。     

胃癌与端粒酶活性表达及DNA倍体关系的研究

目的　揭示胃癌与端粒酶活性及DNA倍体的关系。方法　检测 30例胃癌标本 ,同时取无瘤残端作为对照。端粒酶检测采用端粒重复扩增 酶联免疫吸附法 (TRAP ELISA法 )。DNA倍体的测定采用流式细胞术 ,一步法检测DNA含量。结果　肿瘤瘤体端粒酶阳性率 83 3% (2 5 / 30 ) ,无瘤残端端粒酶阳性率 3 3%(1/ 30 ) (P <0 .0 5 ) ;端粒酶阳性瘤体平均直径 6 5cm ,阴性瘤体平均直径 3 6cm(P <0 .0 5 ) ;端粒酶阳性肿瘤淋巴转移率 5 3 3% (16 / 30 ) ,阴性者无淋巴转移 (0 / 5 ) (P <0 .0 5 ) ;端粒酶阳性肿瘤中异倍体肿瘤占 5 6 0 % (14/2 5 ) ,而端粒酶阴性者无异倍体出现 (P <0 .0 5 )。结论　胃癌端粒酶活性升高 ,端粒酶阳性肿瘤瘤体大 ,淋巴转移率高 ,且异倍体发生率高 ,预后差。提示端粒酶的激活与胃癌的发生发展有密切关系。     

Telomerase expression of malignant epithelial cells correlates with Dukes' stage in colorectal cancer

Background The ribonucleoprotein telomerase has been proposed as a potential prognostic marker for malignancy. Whether telomerase activity of clinical specimens correlates with other clinico‐pathological variables, however, remains controversial. This is at least in part due to the varying contribution that nonmalignant cells will make to the net activity when extracts are assayed for telomerase activity. We therefore designed experiments to assay telomerase activity of isolated malignant cells of primary colorectal cancers. Methods Thirty colorectal cancer and 20 corresponding specimen taken from macroscopically normal regions of the colon were mechanically disaggregated and digested with collagenase, DNase and hyaluronidase. The epithelial cell population was separated using Ber‐EP4 pan‐epithelial antibody and Magnetic Activated Cell Sorting technique. Haematoxylin and eosin staining was used to assess the proportion of recovered epithelial cells which were malignant. Telomerase activity was assayed by the Telomeric Repeat Amplification Protocol, which was quantified by PhosphorImager with Image Quant software. Results Epithelial cells of three of 20 normal mucosa specimens were telomerase positive with weak activity (mean 3.7 TPGs, Total Product Generated, range 1.4–5.1TPGs). In the cancer group the vast majority (>95%) of the epithelial cells recovered were malignant by cytological criteria. Epithelial cells were telomerase positive in all the cancers, with a wide range of telomerase activity values (mean 22.7 TPGs, range 0.19–308 TPGs). Telomerase activity correlated with Dukes' stage (r = 0.52, P = 0.004, Spearman's rank). Conclusions Pathological stage correlates with telomerase activity of the malignant cell population of the primary tumour in colorectal cancer. This suggests that telomerase activity increases during the progression of a cancer and may have implications for the design of anticancer (antitelomerase) agents.     

The clinical implications of telomerase activity in upper tract urothelial cancer and washings

OBJECTIVE: To measure telomerase activity in upper tract urothelial carcinomas (as renal pelvic tumours comprise nearly half of all kidney tumours in Taiwan, a much higher percentage than in other countries) and to determine whether telomerase activity could be used as an additional diagnostic marker in exfoliated cancer cells present in upper tract urothelial washing fluids, thus providing earlier diagnosis and treatment. Materials and methods Telomerase activity was assessed using the telomeric repeat amplification protocol assay in tissue samples from 31 upper tract urothelial carcinomas (from 29 patients). The feasibility of identifying cancer using telomerase activity in exfoliated cancer cells in 17 upper tract urothelial washing samples was also investigated. RESULTS: Telomerase activity was found in 30 (97%) of the 31 upper tract urothelial cancer tissue samples; telomerase activity was detectable in 95% of superficial cancers and in all 11 invasive tumours. The sensitivity of measuring telomerase activity was 100% for grade 1, 93% for grade 2 and 100% for grade 3 tumours. In contrast, telomerase activity was detected in only two (8%) of 26 normal adjacent tissue samples. When the telomerase activity of urothelial washing fluid was compared with that in the corresponding tumours, there was compatible telomerase activity in 15 of the 17 samples. Telomerase activity was more sensitive than voided urine cytology (15%) and washing fluid cytology (53%). In addition, the telomerase activity was high in metastatic lesions. CONCLUSION: Telomerase activity is present in most upper tract urothelial cancer tissues and may be present at an early stage of carcinogenesis. Telomerase activity can be detected in exfoliated cells in urothelial washing fluids in a high proportion of patients with upper tract urothelial cancer. These results suggest that measuring telomerase activity in the exfoliated cancer cells obtained from urothelial washing could be a potentially useful addition to the conventional diagnostic tools used to identify patients with upper tract urothelial carcinoma.     

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。     

Telomerase activity in the human breast

Telomerase is a ribonucleoprotein enzyme which appears to play an important role in carcinogenesis. Telomerase reactivation seems to be associated with immortalization and malignancy. Using a PCR-based assay, we examined telomerase activity in 50 breast tissue specimens, prospectively obtained from 37 women undergoing elective breast surgery. The specimens examined included normal breast (n=13), benign breast lesions (n=5), ductal carcinoma in situ (n=8) and infiltrating ductal carcinoma (n=24). All normal breast, benign breast and DCIS specimens lacked telomerase activity. Sixteen (67%) of 24 infiltrating carcinomas. In infiltrating ductal cancer, there was a statistically significant association between telomerase activity and nodal metastasis. The present results indicate that telomerase activity is associated with acquisition of invasive malignancy in the human breast and may have a role in complementing cytopathological diagnosis. Telomerase activity as a prognostic marker should be included in future validation studies. In DCIS, telomerase activity may be a late event associated with invasion of the basement membrane.     

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。