乳腺癌端粒酶活性的研究

探讨乳腺癌中端粒酶活性的意义及其与腋窝淋巴结状态的关系。方法用建立在多聚酶链反应基础上的端粒重复序列扩增法,对８８例乳腺癌和１６例乳腺良性疾病进行端粒酶活性检测。结果８８例乳腺癌中,７５例,检测到端粒酶活性,而在１６例乳腺良性疾病中仅２例端粒酶阳性,两组的差异有显著性。     

Telomerase is a highly sensitive and specific molecular marker in fine-needle aspirates of breast lesions.

PURPOSE: Telomerase has been detected in a majority of human malignant tumors, making telomerase activity (TA) one key difference between mortal and immortal cells. In this study, we evaluated in blind-trial fashion the association of TA with cytologic and final clinical/pathologic diagnosis in fine-needle aspirates (FNAs) of breast lesions. MATERIALS AND METHODS: In 172 FNAs, including 80 samples that were cytologically malignant, 18 that were atypical but not diagnostic for malignancy, and 74 that were cytologically benign, TA was determined by a modified nonradioactive telomeric repeat amplification protocol (TRAP) assay. Final diagnosis was made by pathologic examination of follow-up surgical material available for all the cytologically malignant samples, a majority of the cytologically atypical samples, and a portion of the cytologically benign samples. RESULTS: TA was detected in 85 of 172 samples. Comparison of the cytologic and histologic diagnoses with TA showed that 80 of 87 samples from patients with breast cancer were telomerase-positive, resulting in a sensitivity of 92%. TA was found in four of five FNAs from carcinomas that were considered cytologically atypical but not diagnostic for malignancy. Eighty of 85 samples from patients with benign breast lesions were telomerase-negative, revealing a specificity of 94%. The five positive cases in this group were all fibroadenomas with low TA. Among the 18 cases with a cytologic diagnosis of atypia, there was a strong positive relationship between TRAP findings and histologic diagnosis. CONCLUSION: The detection of TA in FNAs of breast lesions is a highly sensitive and specific marker of malignancy and may be used as an adjunct in cases with an equivocal cytologic diagnosis.     

Intratumoral oestrone sulphatase activity as a prognostic marker in human breast carcinoma.

Oestrone sulphatase is an important source of local synthesis of biologically active oestrogens in human breast cancer. The oestrone sulphatase enzyme in the particulate fraction of human breast carcinoma was characterised. The Km was 8.91 microM, and the Vmax was 0.022 nmol min-1 mg-1. Oestrone sulphatase activity was detected in 93 of 104 human breast carcinoma samples (89%), and mean activity was 0.041 nmol min-1 mg-1 (range 0-0.399 nmol min-1 mg-1). There was no significant correlation between intratumoral oestrone sulphatase activity and oestrogen receptor status, or with any other prognostic factors. Intratumoral enzyme levels were not associated with time to recurrence or with overall survival time. It thus appears that, although a useful source of intratumoral oestrogens, oestrone sulphatase activity is not of prognostic significance in breast carcinoma.     

Telomerase activity in ductal carcinoma in situ of the breast

Telomerase plays an important role in maintaining the stability of the chromosomes. Activity of telomerase has been detected in proliferating and immortalized cell lines and in a number of malignant tumors including invasive breast cancer. The aim of the study was to examine telomerase activity in ductal carcinoma in situ (DCIS), which is considered to be a precursor lesion of infiltrating breast carcinoma, using a PCR-based telomerase activity protocol (TRAP). We examined 35 samples obtained from histologically confirmed breast biopsies, including 13 normal breast tissues, 11 infiltrating ductal carcinoma (IDC), nine DCIS, and two DCIS with microinvasion. Telomerase activity was demonstrated in 8/9 samples of DCIS, both samples of DCIS with microinvasion, and all but one sample of IDC. Normal breast tissue had no demonstrable telomerase activity. Our results indicate that telomerase is activated frequently in early breast carcinogenesis, although its utilization as a biomarker in DCIS is questionable.     

alphaB-crystallin as a marker of lymph node involvement in breast carcinoma

BACKGROUND: It was found previously that alphaB-crystallin, a small heat-shock protein, was overexpressed in a metastatic variant of the GI101A human breast carcinoma cell line. The objective of the current study was to determine whether the expression of alphaB-crystallin in primary breast carcinomas was associated with lymph node metastasis and survival. METHODS: Expression of alphaB-crystallin was measured in human breast carcinoma cell lines by immunoblotting. Expression in human breast carcinomas was evaluated by immunohistochemical staining of tissue microarrays that contained samples from 317 patients with lymph node-negative breast carcinoma and 291 patients with lymph node-positive breast carcinoma. RESULTS: It was found that alphaB-crystallin was expressed constitutively in certain breast carcinoma cell lines, including those that were capable of metastasizing in immunodeficient mice. Expression of alphaB-crystallin in human tissue samples was associated strongly with lymph node involvement (P < 0.0001; chi-square test) and, to a lesser degree, with high nuclear grade (P = 0.05). Increased intensity of expression was correlated with shorter survival (P = 0.0091; log-rank test). Multivariate analysis indicated that alphaB-crystallin expression was not independent of lymph node status as a predictor of survival. CONCLUSIONS: The data obtained in the current study revealed a strong association between high expression levels of alphaB-crystallin in primary breast carcinoma specimens and lymph node involvement. Further studies will be needed to prospectively elucidate the role of this novel tumor marker as a clinical prognostic marker in local and locally advanced breast carcinoma as well as its potential status as a new target for therapy in patients with breast carcinoma.     

Uroplakin as a marker for typing metastatic transitional cell carcinoma on fine-needle aspiration specimens

BACKGROUND: Immunohistological markers specific for a single type of epithelium are rare. Recently, urothelium tissue-specific genes were cloned. The genes encoded a family of transmembrane proteins, uroplakins, that are expressed only in urothelial mucosa. Using uroplakin antibodies on paraffin-embedded tissue, a previous study demonstrated positive staining in 66% of metastatic transitional cell carcinoma (TCC) cases and negative staining in all other tumors (including breast, ovarian, lung, and gastrointestinal carcinomas) tested. The current study addresses the diagnostic value of uroplakins in conventional fine-needle aspiration (FNA) material in establishing a diagnosis of metastatic TCC. METHODS: Representative slides from 27 FNA cases of metastatic TCC and 52 non-TCC carcinomas were collected. The avidin-biotin-peroxidase method was utilized, using polyclonal antiuroplakin as the primary antibody on 95% ethanol-fixed, Papanicoloau-stained direct smears. RESULTS: Twenty-five of 27 metastatic TCC cases (93%) were found to stain positively for uroplakin with a superficial membrane/microluminal staining pattern. A few cells with diffuse membranous staining also were noted in 48% of the positive metastatic TCC cases. The superficial membrane/microluminal staining pattern was not observed in any of the non-TCC carcinomas. However, approximately 6% of these cases (3 of 52 cases) did show rare tumor cells with diffuse membranous staining. CONCLUSIONS: The application of uroplakin antibodies to 95% ethanol-fixed FNA direct smears has improved the sensitivity of the antibody for metastatic TCC while maintaining a specificity comparable to that of paraffin-embedded tissue. The authors believe that these antibodies have diagnostic potential in cytopathology in the evaluation of metastatic TCC.     

乳腺癌患者外周血中端粒酶活性表达及其临床意义

目的：探讨端粒酶活性在乳腺癌患外周血癌细胞中的表达及其与浸润转移的关系。方法：通过用TRAP-PCR-ELISA法检测65例乳腺癌患外周血端粒酶的活性，按照肿瘤大小、分级程度、浸润情况、淋巴结有无转移来分析端粒酶与乳腺癌患生物学行为的相关性。结果：外周血中检测端粒酶活性阳性率为40．00％，低于相应的乳腺癌组织阳性率(61．54％)，乳腺癌患外周血中肿瘤细胞端粒酶相对活性的升高与患的淋巴结转移和浸润呈正相关。结论：检测患外周血中端粒酶活性可能优于检测肿瘤组织的端粒酶活性，可以对肿瘤患的病情进行实时监控，指导临床化疗。     

Telomerase activity in ductal carcinoma in situ and invasive breast cancer.

The increasing number of breast carcinoma in situ detected by screening procedures makes it imperative to develop improved markers to stratify the risk of invasive cancer. Telomerase is detectable in invasive cancer, but not in normal tissues. We have microdissected frozen tissue blocks containing both DCIS and invasive cancer to assay the telomerase activity of these two lesions. The 46 available cases of concurrent DCIS and invasive breast cancer resulted in 43 DCIS samples and 38 invasive cancer samples adequate for analysis. Seventy per cent of the DCIS and all invasive cancer samples tested had detectable telomerase activity. In addition, we analysed telomerase activity in ten cases of DCIS that were not associated with invasive cancer, and detected telomerase activity in seven (70%). Mixing experiments showed no evidence of telomerase inhibitors in telomerase negative samples. Furthermore, periductal inflammatory infiltrates were shown to be a potential confounding source of telomerase activity. Since DCIS lesions appear to be heterogeneous with respect to telomerase activity, and telomerase activation appears to precede the development of invasive cancer, telomerase activity may be a useful adjunct in stratifying the risk of developing invasive breast cancer in patients with DCIS.     

The use of CA15.3 as a serum tumour marker in breast carcinoma

There is, as yet, no tumour marker which is sufficiently specific and sensitive for use in the routine assessment of breast cancer patients. CA15.3 is a recently described tumour marker determined by two monoclonal antibodies. We have estimated CA15.3 by immunoradiometric assay in 187 patients attending a breast clinic. Eighty-one patients with benign disease were used as controls and 32 U/ml was taken as the upper limit of the normal range (means + 3SD = 31.7). Of 58 women with Stage I and II disease, only four had abnormal concentrations of CA15.3 and all are disease-free at a mean follow-up of 31 months. Seven women with normal CA15.3 concentrations developed recurrent disease at a mean of 18.7 months (range 10-25 months). Seven-day postoperative values were significantly lower than pre-operative values. There was no association between the CA15.3 value and the axillary nodal status. The patients with disseminated disease had a wide range of CA15.3 concentrations and there was no association between the CA15.3 concentration and the apparent tumour load.     

原发性肺癌与食管癌组织中端粒酶活性表达的临床研究

目的 :探讨端粒酶作为一种新的生物学标志物用于常见胸部恶性肿瘤诊断和预后评价等方面的临床应用价值。方法 :采用端粒重复序列扩增法对原发性肺癌、食管癌手术切除组织及相应的癌旁组织进行端粒酶活性分析 ,并以相应的良性病变作为对照。结果 :3 2例肺癌组织中端粒酶阳性率为 84 4%( 2 7/3 2 ) ,癌旁组织为 9 4% ( 3 /3 2 ) ,良性组织中未检出端粒酶阳性。端粒酶活性随肺癌临床分期有升高趋势 ,伴淋巴结转移标本组阳性率 94 7% ( 18/19)明显高于非淋巴结转移组69 2 % ( 9/13 ) ,P <0 0 5 ;但与肿瘤病理组织学类型、分化程度等差异无统计学意义 ,P >0 0 5。 3 7例食管癌端粒酶阳性率为 86 5 % ( 3 2 /3 7) ,癌旁组织为 18 9% ( 7/3 7) ,正常食管组织未检出端粒酶阳性。当癌侵及肌层及外侵后 ,其阳性表达率 10 0 % ( 2 3 /2 3 )高于局限于黏膜及黏膜下层者 71 4% ( 10 /14 ) ,P <0 0 1。伴淋巴结转移标本阳性率 10 0 % ( 2 1/2 1)高于非淋巴结转移者 75 % ( 12 /16) ,P <0 0 5。结论 :端粒酶有可能成为肺癌、食管癌早期诊断和判断预后的重要生物标志物和治疗新靶点。     

子宫内膜癌端粒酶活性研究

目的探索端粒酶活性与子宫内膜癌发生发展的关系。方法采用ＰＣＲ－ＴＲＡＰ方法检测３４例子宫内膜癌和６例正常子宫内膜组织的端粒酶活性。结果在３４例子宫内膜癌中,２８例端粒酶活性阳性;６例正常子宫内膜组织中,有１例端粒酶活性阳性。结论端粒酶激活与子宫内膜癌的发生发展有密切关系,并有可能成为子宫内膜癌的临床肿瘤标记物     

Telomerase activity in transthoracic fine-needle biopsy aspirates from non-small cell lung cancer as prognostic factor of patients' survival

PURPOSE: Evaluation of the relationship between telomerase activity in transthoracic fine-needle biopsy (TFNB) aspirates collected from peripheral non-small cell lung cancer (NSCLC), cancer advancement, risk of death and free of cancer recurrence survival. MATERIAL AND METHODS: The study group consisted of 93 patients with peripheral infiltration of the lung. All of them had TFNB of the focal pulmonary lesion performed. The aspirates were subjected to standard cytological evaluation. Telomerase activity in the specimens was determined with the PCR-ELISA PLUS method. Cancer advancement, manner of treatment and survival were assessed in patients with NSCLC. RESULTS: A benign lesion of the lung was recognised in 14 cases. NSCLC was newly diagnosed in 79 subjects. Nobody with benign infiltration had a detectable level of telomerase in lung infiltration. Increased telomerase activity was observed in 56 (70.9%) patients with lung cancer. It was significantly more often detected in patients with non-operable NSCLC (clinical stage IIIB plus IV) and patients with distant metastases (stage IV alone). Cox hazard analysis revealed that presence of telomerase activity in primary NSCLC is an independent prognostic factor of survival. Increased telomerase activity in TFNB aspirates was related to 7 times higher relative risk of death during the study [RR=6.9 (CI: 1.8-26.8); p<0.05] and 2.5 times higher risk of cancer recurrence after radical treatment [RR=2.5 (CI: 0.3-9.3); p<0.05]. CONCLUSION: Telomerase activity in aspirates collected through TFNB of primary peripheral NSCLC could be a helpful independent prognostic factor of distant metastases and risk of death or cancer recurrence.     

Telomerase activity in stage II colorectal carcinoma

BACKGROUND: Telomerase is a ribonucleoprotein polymerase that adds telomeric repeats to chromosome ends. This enzyme is deficient in the majority of normal somatic cells, but often is reactivated during tumorigenesis. In the current study, the authors examined telomerase activity in human American Joint Committee on Cancer Stage II colorectal carcinomas and correlated it with traditional prognostic indicators and disease outcome. METHODS: The telomerase repeat amplification protocol (TRAP) was employed to determine telomerase activity in 122 surgical specimens (from 77 male and 45 female patients) of human Stage II colorectal carcinoma. The primary site of the tumor was the colon in 52 cases and the rectum in 70 cases. Telomerase activity was correlated with traditional prognostic indicators such as gender, age, T classification, tumor size, tumor grade, and disease outcome (overall survival and disease-free survival). The Median follow-up for patients who still were alive was 5.8 years. RESULTS: Telomerase activity was detected in 80% of the tumors (98 of 122 tumors). Telomerase-positive patients differed from telomerase-negative patients in that they tended to be female (41% vs. 21%; P = 0.1), presented with primary tumors of the colon more frequently (49% vs. 17%; P = 0.01), and had a higher T classification (T(4)) (62% vs. 38%; P = 0.04). Univariate and multivariate analyses demonstrated a correlation between telomerase activity and disease-free survival (P = 0.05). CONCLUSIONS: Although a large percentage of Stage II colorectal carcinoma samples were positive for telomerase activity, the prognosis for patients with telomerase-negative tumors was found to be worse than that for patients with telomerase-positive tumors.     

Telomerase activity as a prognostic factor in colorectal cancer

BACKGROUND: As the enzyme telomerase extends the life of the cell through its ability to lengthen telomeres, its activity in different types of tumor has been evaluated as a possible factor involved in tumorigenesis. The aim of this study was to assess the prognostic significance of telomerase activity in patients with colorectal carcinoma. PATIENTS AND METHODS: Telomerase activity was determined in 103 patients undergoing surgery for colorectal cancer between 2001 and 2003. Telomerase activity was determined by an enzyme-linked immunoassay based on the amplification of telomeric repeat sequences (TRAP assay). RESULTS: 90% of our study population showed telomerase activity. Telomerase activity was related to tumor stage and site: a lower proportion of patients with stage A tumors showed telomerase activity compared to more advanced stages; and more patients with colon than with rectal carcinomas were telomerase positive. Multivariate analysis revealed that by adjusting for tumor stage, telomerase activity could be used to predict the risk of death or recurrence (p < 0.001). CONCLUSIONS: Activation of telomerase seems to be a frequent event related to the stage of the tumor in colorectal tumorigenesis. Our findings suggest that telomerase activity can predict a greater risk of death or recurrence, irrespective of the more conventional prognostic factors.