Invasion factors uPA/PAI-1 and HER2 status provide independent and complementary information on patient outcome in node-negative breast cancer.

PURPOSE: The independent clinical relevance of invasion factors urokinase-type plasminogen activator (uPA)/PAI-1 and HER2 status was evaluated in lymph node-negative breast cancer patients (N = 118) without adjuvant systemic therapy after long-term follow-up of more than 10 years (median, 126 months). PATIENTS AND METHODS: Levels of uPA and its inhibitor PAI-1 were prospectively measured by enzyme-linked immunosorbent assay in primary tumor tissue extracts. HER2 gene amplification (HER2_AMP) was evaluated by fluorescence in situ hybridization (FISH; Ventana Medical Systems HER-2/neu probe; Tucson, AZ), and HER2 protein overexpression (HER2_EXP) was evaluated by immunohistochemistry (IHC; Oncogene Science antibody Ab-3; Cambridge, MA) on parallel-cut formalin-fixed paraffin-embedded tissue sections. RESULTS: uPA/PAI-1 was high (either one or both factors were high) in 44% of the tumors. HER2_AMP was detected by FISH in 33% of the patients, and HER2_EXP was found by IHC in 44% of the patients. In a multivariate analysis of established and tumor-biologic prognostic factors, uPA/PAI-1 was the only independent prognostic factor for disease-free survival ([DFS]; P <.001; relative risk [RR], 8.3; 95% confidence interval [CI], 3.4 to 20.4). Although HER2_AMP and HER2_EXP did not reach significance for DFS, they were significant for overall survival (OS), even in multivariate analysis (HER2_AMP: P =.004; RR, 3.7; 95% CI, 1.5 to 9.2; HER2_EXP: P =.009; RR, 3.4; 95% CI, 1.4 to 8.7). CONCLUSION: After long-term follow-up, uPA/PAI-1 levels in primary tumor tissue reliably and strongly indicate an aggressive course of disease in lymph node-negative breast cancer independent of HER2 status. The particular prognostic effect of HER2 status on OS may reflect its ability to predict resistance to systemic therapy.     

Risk-group discrimination in node-negative breast cancer using invasion and proliferation markers: 6-year median follow-up.

Factors reflecting two major aspects of tumour biology, invasion (urokinase-type plasminogen activator (uPA), plasminogen activator inhibiter (PAI-1), cathepsin D) and proliferation (S-phase fraction (SPF), Ki-67, p53, HER-2/neu), were assessed in 125 node-negative breast cancer patients without adjuvant systemic therapy. Median follow-up time was 76 months. Antigen levels of uPA, PAI-1 and cathepsin D were immunoenzymatically determined in tumour tissue extracts. SPF and ploidy were determined flow-cytometrically, Ki"'-67, p53, and HER-2/neu immunohistochemically in adjacent paraffin sections. Their prognostic impact on disease-free (DFS) and overall survival (OS) was compared to that of traditional factors (tumour size, grading, hormone receptor status). Univariate analysis determined PAI-1 (P < 0.001), uPA (P = 0.008), cathepsin D (P = 0.004) and SPF (P = 0.023) as significant for DFS. All other factors failed to be of significant prognostic value. In a Cox model, only PAI-1 was significant for DFS (P < 0.001, relative risk (RR) 6.2). In CART analysis for DFS, the combination of PAI-1 and uPA gave the best risk group discrimination. For OS, PAI-1, cathepsin D, tumour size and ploidy were statistically significant in univariate, but PAI-1 was the only independently significant factor in Cox analysis (P < 0.001, RR 8.9). In particular, this analysis shows that PAI-1 is still a strong and independent prognostic factor in node-negative breast cancer after extended 6-year median follow-up.     

Survivin expression and targeting in breast cancer

IntroductionSurvivin a multifunctional protein that controls cell division, inhibition of apoptosis and promotion of angiogenesis. It is expressed in most human neoplasm, but is absent in normal and differentiated tissues. The purpose of this article is to overview the expression of survivin, effect of its expression in response to treatment, correlation with other markers and newer advancement in targeting survivin.MethodsA detailed search of Medline was carried out using the following search strategy: “((survivin) OR ((apoptosis) AND (inhibitor OR inhibitors))) AND ((breast) AND (neoplasm OR neoplasms OR tumor OR tumor OR cancer OR carcinoma))”. Abstract of all articles thus identified were reviewed to identify the relevant studies, full articles of studies thus identified were then obtained and reviewed. All relevant data was extracted and tabulated.ResultsSurvivin expression by Immunohistochemistry was identified in 65.3% (55.2–90.0%) of the breast cancer patients among the identified studies while survivin mRNA by RT-PCR was identified in 93.6% (90–97%). Survivin expression has been reported to be associated with over expression of HER 2, vascular endothelial growth factor (VEGF), urokinase plasminogen activator (uPA)/PAI-1.ConclusionSurvivin is over expressed in majority of breast cancers. The over expression of survivin is found to correlate with HER 2 and EGFR expression. Survivin expression has been found to confer resistance to chemotherapy and radiation. Targeting survivin in experimental models improves survival. More studies are needed on the role of survivin in multi drug resistance (MDR) in the presence of Pgp/uPA/PAI-1 and the impact of survivin over expression in triple negative breast cancer.     

Prognostic impact of proteolytic factors (urokinase-type plasminogen activator, plasminogen activator inhibitor 1, and cathepsins B, D, and L) in primary breast cancer reflects effects of adjuvant systemic therapy.

PURPOSE: Prognostic and predictive impact of five proteolytic factors associated with tumor invasion and metastasis in primary breast cancer were evaluated after long-term follow-up. EXPERIMENTAL DESIGN: Antigen levels of urokinase-type plasminogen activator, plasminogen activator inhibitor-1 (PAI-1), Cathepsins B, D, and L were determined using immunochemical assays in primary tumor tissue of 276 patients. RESULTS: During follow-up (median 109 months), 119 (43%) patients relapsed, and 117 (42%) died. In the whole collective, lymph node status (P < 0.001; RR 3.8), Cathepsin L (P < 0.001; RR 2.6), and PAI-1 (P = 0.027; RR 1.7) were the only independent significant factors in multivariate analysis for disease-free survival (DFS). For overall survival (OS), lymph node status (P < 0.001; RR 2.9), Cathepsin L (P = 0.017; RR 1.9), PAI-1 (P = 0.01; RR 1.9), and grading (P = 0.026; RR 1.7) were significant. In the node-negative subgroup, PAI-1 was the only significant factor for DFS (P = 0.004; RR 3.7) and the strongest factor (P = 0.004; RR 3.7) for OS next to grading (P = 0.017; RR 3.1). In node-positive patients, Cathepsin L was the only significant factor for both DFS (P < 0.001; RR 3.2) and OS (P = 0.003; RR 2.5). For all proteolytic factors but Cathepsin L, the univariate prognostic impact on DFS was substantial in patients without adjuvant systemic therapy but was diminished if adjuvant therapy had been administered. Cathepsin L maintained its strong prognostic impact on DFS even in patients with adjuvant endocrine therapy (P = 0.01; RR 2.8). CONCLUSIONS: The observed effect of adjuvant systemic therapy on their prognostic strength suggests that the assessed proteolytic factors supply predictive information on therapy response.     

Her-2/neu and urokinase-type plasminogen activator and its inhibitor in breast cancer.

PURPOSE: Recent studies suggest that HER-2/neu specifically promotes the invasive capacity of tumor cells by up-regulating secretion of the proteolytic enzyme, urokinase-type plasminogen activator (uPA), or its inhibitor, plasminogen activator inhibitor-1 (PAI-1), in colon and gastric cancer. It was the purpose of this study to: (a) evaluate the association between HER-2/neu and uPA and PAI-1 expression in a large primary breast cancer cohort; (b) perform the first multivariate analysis, including HER-2/neu, uPA, and PAI-1 in breast cancer; and (c) define the effect of HER-2/neu overexpression on uPA and PAI-1 expression in breast cancer cells. EXPERIMENTAL DESIGN: HER-2/neu, uPA, and PAI-1 were measured as continuous variables by ELISA in primary breast cancer tissue extracts from 587 patients with clinical follow-up and analyzed for correlations with clinical outcome. Furthermore, a full-length human HER-2/neu cDNA was introduced into five human breast cancer cell lines to define the effects of HER-2/neu overexpression on uPA and PAI-1 expression. In addition, we tested whether HER-2/neu antibodies could reverse any given alteration of uPA and PAI-1 levels. RESULTS: Our findings indicate a weak positive association between HER-2/neu and uPA (r = 0.147; P < 0.001) and no association between HER-2/neu and PAI-1 (r = 0.07; P = 0.085). HER-2/neu overexpression (> or =400 fmol/mg) and high levels of uPA/PAI-1 (> or =5.5 ng/mg and/or > or =14 ng/mg, respectively) were significantly associated with shorter disease-free survival (DFS; P < 0.001 and P = 0.003) and metastasis-free survival (MFS; P = 0.015 and P < 0.001). Multivariate analysis revealed prognostic independence between HER-2/neu and the uPA/PAI-1 axis for DFS and MFS. Both uPA and PAI-1 had no significant discriminatory effect among HER-2/neu-positive patients for DFS. The prognostic value of HER-2/neu overexpression for MFS, however, was significantly enhanced by elevated uPA expression (P = 0.053). Stable transfection of the HER-2/neu gene into multiple human breast cancer cell lines resulted in consistent down-regulation of uPA or PAI-1 expression. In addition, anti-HER-2/neu antibodies did not significantly affect uPA or PAI-1 expression in human cancer cell lines naturally overexpressing HER-2/neu. CONCLUSIONS: The present findings suggest that the invasive phenotype elicited by HER-2/neu overexpression in breast cancer is not a direct effect of uPA or PAI-1 expression. HER-2/neu and the uPA/PAI-1 axis have been shown to affect the invasive capacity of breast cancer independently. Determination of uPA can provide significant additional prognostic information for MFS in HER-2/neu-positive and -negative patients.     

Hypoxia-inducible factor 1α is closely linked to an aggressive phenotype in breast cancer

PURPOSE: The aim of this study is to clarify characteristics of invasive breast cancer with expression of Hypoxia-induced factor 1alpha (HIF-1alpha) which is induced by hypoxia and signal transduction of growth factors. EXPERIMENTAL DESIGN : We examined, by immunohistochemical analysis, the expression of HIF-1alpha in normal breast tissue, benign disorders and breast cancer. In invasive breast cancer, we investigated the correlation between expression of HIF-1alpha and clinicopathological and biological factors. We also studied the prognostic value of HIF-1alpha in breast cancer. RESULTS : HIF-1alpha was mainly detected in tumor cell nuclei. In the 171 cases of invasive breast cancer examined, nuclear HIF-1alpha expression was detected in 63 (36.8%) cases. Immunoreactive nuclear HIF-1alpha was correlated with tumor size (p = 0.0013), lymph node metastasis (p = 0.0005), tumor stage (p = 0.0031) and histological grade (p = 0.0074). Elevated HIF-1alpha levels was also associated with hormone receptor negativity (p = 0.0025), HER2 overexpression (p = 0.0053), high Ki67 labeling index (p = 0.0002), increased levels of VEGF (p < 0.0001), COX-2 overexpression (p = 0.0029) and increased nuclear p53 (p = 0.0048). In terms of the possible use of HIF-1alpha as a prognostic indicator, patients who had increased HIF-1alpha levels in their tumor showed shorter disease-free survival (DFS) (p < 0.0001) and overall survival (OS) (p = 0.0002) than those lacking HIF-1alpha in univariate analysis. In multivariate analysis of DFS and OS, HIF-1alpha was identified an independent prognostic factor. CONCLUSIONS: These findings suggest that HIF-1alpha was closely linked to an aggressive phenotype in breast cancer. It may be useful to study the expression of HIF-1alpha using immunohistochemical analysis for better understanding of the tumor characteristics of breast cancer.     

Association between intratumoral free and total VEGF, soluble VEGFR-1, VEGFR-2 and prognosis in breast cancer

Vascular endothelial growth factor (VEGF) receptors consist of three cell-membrane type receptors (VEGFR-1, VEGFR-2 and VEGFR-3), and soluble form of VEGFR-1 (sVEGFR-1), an intrinsic negative counterpart of the VEGF. In this study, we measured intratumoral protein levels of free and total VEGF, VEGFR-2 and sVEGFR-1 from 202 primary breast cancer tissues and examined their prognostic values. A significant inverse correlation was found between free or total VEGF and oestrogen receptor (ER) status (P=0.042 and 0.032, respectively). A univariate analysis showed that low sVEGFR-1 and high total VEGF were significantly associated with poor prognosis in disease-free survival (DFS) and overall survival (OS). The ratio of sVEGFR-1 to total VEGF was a strong prognostic indicator (DFS: P=0.008; OS: P=0.0002). A multivariate analysis confirmed the independent prognostic values of total VEGF and the ratio of sVEGFR-1 to total VEGF. In subgroup analysis, total VEGF was a significant prognostic indicator for ER-positive tumours but not for ER-negative tumours, whereas sVEGFR-1 was significant for ER-negative tumours but not for ER-positive tumours. In conclusion, the intratumoral sVEGFR-1 level, VEGF level and the ratio of sVEGFR-1 to total VEGF are potent prognostic indicators of primary breast cancer, and might be relevant to ER status.     

Prognostic value of uPA,PAI-1 and PAI-2 mRNA expression in primary breast cancer

BACKGROUND: The prognostic value of uPA, PAI-1 and PAI-2 mRNA expression was studied in a retrospective series of 130 primary breast cancer patients (median follow-up 8.1 years). MATERIALS AND METHODS: UPA, PAI-1 and PAI-2 mRNA were quantified by means of real-time quantitative RT-PCR. Comparison with the corresponding protein levels determined by ELISA was performed in 21 cases. RESULTS: Higher uPA protein values were found in cases with high mRNA values, but the relationship was of borderline significance. PAI-1 and PAI-2 mRNA were positively correlated to protein values. The mRNA expression of uPA, PAI-1 and PAI-2 was significantly correlated with one another. Higher uPA and PAI-1 mRNA values were significantly associated with shorter disease-free survival (DFS) (p = 0.002; p = 0.03, respectively). Low and very high PAI-2 mRNA values tended to be associated with longer DFS. In Cox multivariate analyses, higher uPA and PAI-1 mRNA values were independently associated with shorter DFS. PAI-2 was not retained as a significant variable in the Cox model. CONCLUSION: These preliminary results indicate that, in breast cancer, uPA, PAI-1 and PAI-2 mRNA analysis by quantitative RT-PCR give results comparable to those obtained at the protein level.     

Time-varying prognostic impact of tumour biological factors urokinase (uPA), PAI-1 and steroid hormone receptor status in primary breast cancer.

In breast cancer, several investigations have demonstrated that the tumour biological factors uPA urokinase-type plasminogen activator) and its inhibitor PAI-1 are statistically independent, strong prognostic factors for disease-free (DFS) and overall survival (OS). However, statistical analyses performed for varying follow-up periods suggested a time variation of prognostic strength. We therefore investigated the time-dependent prognostic power of uPA, PAI-1 and steroid hormone receptor status applying the time-varying coefficient model of Gray. uPA and PAI-1 were analysed by enzyme-linked immunosorbent assay in tumour tissue extracts from 314 breast cancer patients. Hormone receptors (oestrogen and progesterone) were determined by radioligand binding or by immunohistochemistry. Univariate and multivariate analyses (Cox proportional hazards model) of DFS and OS were performed for all patients, including 147 node-negative patients. Median follow-up of patients still alive at time of analysis (n = 232) was 58 months. Although initially of high prognostic impact, a continuous decrease over time in the prognostic power of hormone receptor status and uPA was observed. In contrast, the prognostic impact of PAI-1 increased over time and reached similar strength as the lymph node status. The time-dependent risk profile of prognostic factors may have important clinical implications in regard to follow-up and patients'' individual risk situation. Evaluation of time dependency of prognostic factors may also give a more profound insight into the dynamics of breast cancer metastasis.     

乳腺癌中GRP78、Survivin表达的相关性及与预后的关系

目的:探讨乳腺癌中GRP78、Survivin表达的相关性及其与乳腺癌患者预后的关系.方法:采用免疫组化Envision法检测169例乳腺癌石蜡标本中GRP78、Survivin的表达状况,并对其表达的相关性以及与乳腺癌患者预后的关系进行了统计分析.结果:在169例乳腺癌标本中,GRP78强表达为68例(40.2%),弱表达为101例(59.8%);Survivin强表达为96例(56.8%),弱表达为73例(43.2%).对二者相关性的分析表明二者表达的吻合度具有统计学意义,但吻合程度较弱.单因素生存分析结果表明GRP78(+)Survivin(+)组(1组)、GRP78(-)Survivin(+)组(2组)、GRP78(+)Survivin(-)(3组)之间的无病生存时间差异无显著性(P>0.05),但与GRP78(-)Survivin(-)组(4组)相比,其无病生存时间均较差,差异有显著性(P<0.05);1、2、3组间的总生存时间差异无显著性(P>0.05),但与4组相比,其总生存时间均较差,差异有显著性(P<0.05).多因素Cox回归分析表明GRP78、Survivin的表达状况是影响乳腺癌患者无病生存时间的独立预后因素(P<0.05),但不是总生存时间的独立预后因素(P>0.05).结论:乳腺癌中GRP78与Survivn存在着相关性,GRP78和/或Survivn强表达提示患者预后不良,GRP78、Survivin的表达状况对判断乳腺癌患者的预后具有一定的参考价值.     

Altered cytoplasmic-to-nuclear ratio of survivin is a prognostic indicator in breast cancer.

PURPOSE: Survivin (BIRC5) is a promising tumor biomarker. Conflicting data exist on its prognostic effect in breast cancer. These data may at least be partly due to the manual interpretation of immunohistochemical staining, especially as survivin can be located in both the nucleus and cytoplasm. Quantitative determination of survivin expression using image analysis offers the opportunity to develop alternative scoring models for survivin immunohistochemistry. Here, we present such a model. EXPERIMENTAL DESIGN: A breast cancer tissue microarray containing 102 tumors was stained with an anti-survivin antibody. Whole-slide scanning was used to capture high-resolution images. These images were analyzed using automated algorithms to quantify the staining. RESULTS: Increased nuclear, but not cytoplasmic, survivin was associated with a reduced overall survival (OS; P = 0.038) and disease-specific survival (P = 0.0015). A high cytoplasmic-to-nuclear ratio (CNR) of survivin was associated with improved OS (P = 0.005) and disease-specific survival (P = 0.05). Multivariate analysis revealed that the survivin CNR was an independent predictor of OS (hazard ratio, 0.09; 95% confidence interval, 0.01-0.76; P = 0.027). A survivin CNR of >5 correlated positively with estrogen receptor (P = 0.019) and progesterone receptor (P = 0.033) levels, whereas it was negatively associated with Ki-67 expression (P = 0.04), p53 status (P = 0.005), and c-myc amplification (P = 0.016). CONCLUSION: Different prognostic information is supplied by nuclear and cytoplasmic survivin in breast cancer. Nuclear survivin is a poor prognostic marker in breast cancer. Moreover, CNR of survivin, as determined by image analysis, is an independent prognostic factor.     

Association of urokinase-type plasminogen activator and its inhibitor with disease progression and prognosis in ovarian cancer.

PURPOSE: Urokinase-type plasminogen activator (uPA) and its inhibitor, plasminogen activator inhibitor (PAI)-1, have been shown to be related to poor prognosis in a variety of malignant solid tumors. Studies on the prognostic relevance of uPA and PAI-1 in ovarian cancer, however, have been inconclusive. The current study tests the hypothesis that elevated expression of uPA and PAI-1 is associated with prognosis and disease progression. EXPERIMENTAL DESIGN: uPA and PAI-1 were prospectively measured by quantitative ELISA in tumor samples from 103 ovarian cancer patients (82 primary invasive epithelial carcinomas, 9 low malignant potential tumors, and 12 recurrent ovarian carcinomas). RESULTS: uPA but not PAI-1 levels were consistently associated with malignant progression, with levels increased from low malignant potential tumors to primary tumors (uPA, P = 0.04; PAI-1, P = 0.019), from early to advanced disease stages (uPA, P = 0.014; PAI-1, P = 0.23), and from primary to intra-abdominal metastatic tumors (uPA, P = 0.001; PAI-1, P = 0.16). High uPA and PAI-1 levels were associated with residual tumor volumes of >1 cm (P = 0.001 and P = 0.016, respectively). Among invasive International Federation of Gynecologists and Obstetrician stages I-IV tumors, elevated levels of uPA (>5.5 ng/mg) and PAI-I (>18.8 ng/ml) were associated with a shortened progression-free survival (uPA, P = 0.003; PAI-1, P = 0.039) and overall survival (uPA, P = 0.0002; PAI-1, P = 0.007). In multivariate analysis, uPA retained prognostic independence for progression-free survival (P = 0.037) and overall survival (P = 0.006). CONCLUSIONS: These data suggest that the uPA/PAI-1 axis may play an important role in the intra-abdominal spread and reimplantation of ovarian cancer cells. The prognostic relevance of uPA and PAI-1 supports their possible role in the malignant progression of ovarian cancer.     

Prognostic significance of PAI-1 and uPA in cytosolic extracts obtained from node-positive breast cancer patients

Cancer cell invasion is accomplished by the concertedaction of several extracellular proteolytic enzyme systems, oneof which is the urokinase plasminogen activation system.The different components of this system, e.g. urokinaseplasminogen activator (uPA), its receptor uPAR, as wellas its main inhibitor plasminogen activator inhibitor type1 (PAI-1) have all been shown to haveprognostic value in breast cancer, i.e. high tumorlevels are associated with a poor prognosis.In orderto further substantiate the prognostic value of uPAand PAI-1, we have tested the cutpoints (medianvalues and optimized cutpoints) from our first study(Cancer Res 53: 2513–2521, 1993) in an independentgroup of breast cancer patients. Breast cancer cytosolsfrom 100 premenopausal and 150 post-menopausal node positivepatients were included. The median observation time was80 months (range 49–145). Univariate analysis showed thathigh PAI-1 levels (above the median PAI-1 value)were significantly associated with short recurrence-free survival (RR:1.65; 95% CI: 1.04–2.63; P=0.03) andshort overall survival (RR: 2.46; 95% CI: 1.52–3.96;P=0.0001) in postmenopausal patients. Postmenopausal patientswith high uPA levels (above the median uPAvalue) had a significantly shorter recurrence-free survival (RR:2.04; 95% CI: 1.17–3.56; P=0.01) andoverall survival (RR: 2.07; 95% CI: 1.16–3.70; P= 0.01) than patients with low uPA values.Nearly identical results were obtained when using theoptimized PAI-1 or uPA value.In a Cox multivariateanalysis which included other established prognostic factors, highPAI-1 was found to be an independent prognosticvariable predicting short overall survival with a relativerisk of 2.27 in postmenopausal women, and highuPA was found to be an independent prognosticvariable predicting short recurrence-free survival with a relativerisk of 1.86 in postmenopausal women. The presentstudy indicates that uPA and PAI-1 are independentand significant prognostic variables in subsets of breastcancer patients.     

Survivin在乳腺癌组织中的表达及其临床意义

目的:探讨Survivin在乳腺癌组织中的表达与凋亡指数(apoptosis index,AI)以及临床指标的相关性和预后的意义。方法:免疫组化法检测63例乳腺癌标本中Survivin的表达;TUNEL法检测乳腺癌组织中的AI。结果:Survivin在乳腺癌组织中的表达阳性率为63·5%。Survivin与AI呈负相关,P=0·0000。Survivin与肿块直径、肿瘤分期、雌激素受体状态无关,而与腋窝淋巴结转移呈正相关,P=0·0000。单因素分析显示,Survivin是乳腺癌的一个预后指标,P=0·0115。结论:Survivin是一种有效的凋亡抑制蛋白,在肿瘤的发生过程中起一定作用。检测Survivin对乳腺癌的预后分析和进一步治疗具有指导作用。肿瘤防治杂志,2005,12(19):1476-1479     

Prognostic correlation of basic fibroblast growth factor and vascular endothelial growth factor in 1307 primary breast cancers

This study was designed to investigate the possible relationship between the protein expression of basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) with p53 status, breast cancer prognostic factors, metastatic site, and survival after adjuvant therapy. Basic fibroblast growth factor and VEGF expression were determined by enzyme-linked immunosorbent assays in cytosol specimens obtained from 1307 patients with T1-3 primary breast cancer (789 node-negative, 518 node-positive) diagnosed between 1990 and 1997. The median follow-up time was 70 months. Increased bFGF expression was more frequently found in tumors with low VEGF expression (r = -0.286; P = 0.095). Increased bFGF was associated with smaller tumors (P < 0.001), absence of axillary metastasis (P = 0.003), low S-phase fraction (P < 0.001), and longer recurrence-free survival (RFS; P = 0.0038) and overall survival (OS; P = 0.0316). Vascular endothelial growth factor was a prognostic factor for RFS (P < 0.0001) and OS (P < 0.0001) in univariate and multivariate analyses (RFS: 95% CI, 1.1-1.7; P = 0.036; OS: 95% CI, 1.2-2.2; P = 0.002), whereas bFGF expression was not correlated with RFS or OS. Increased VEGF content was correlated with shorter survival after adjuvant endocrine therapy (RFS, P = 0.0004; OS, P = 0.0009). Patients with estrogen receptor-negative disease were excluded from the analysis. Basic fibroblast growth factor was not a prognostic factor after adjuvant systemic therapy, nor was it related to metastatic site. Expression of VEGF is an independent prognostic factor for patients with primary breast cancer. High bFGF expression was related to good prognostic features and longer survival times, but did not add prognostic information in multivariate analysis. The results might implicate that different angiogenic pathways exist in human breast cancer.     

Prognostic value of the urokinase-type plasminogen activator,and its inhibitors and receptor in breast cancer patients

Urokinase-type plasminogen activator (uPA), its inhibitors (PAI-1 and PAI-2), and its receptor (uPAR) play a key role in tumor invasion and metastasis. This study was designed to evaluate the prognostic impact of uPA, PAI-1, PAI-2, and uPAR and the combination of these factors in a group of 460 primary breast cancer patients. Concentrations of all 4 components of the uPA system were measured in tumor extracts using enzyme-linked immunosorbent assays (American Diagnostica, Inc, Greenwich, CT). After a median follow-up of 33 months, 18.5% of the patients had relapsed. The Cox proportional hazards model was applied for both univariate and multivariate analyses of disease-free survival (DFS). PAI-1 and PAI-2 were shown to provide independent prognostic information in breast cancer. Patients with either low levels of PAI-1 or high levels of PAI-2 were found to have better DFS (relative risk was 2.08 and 1.78, respectively). The prognostic value could be even further improved by a combination of both inhibitors. Aside from the uPA inhibitors, only nodal status and hormonal receptor status retained independent prognostic value. The other 2 invasion markers, uPA and uPAR, showed no statistically significant impact on DFS. In our patients, who were mostly treated with adjuvant therapy, uPA was not found to be an independent prognostic marker for DFS; this could be a consequence of the predictive value of uPA for response to adjuvant therapy and should be further investigated.     

乳腺癌组织中survivin、c-erbB-2基因的表达与预后的关系

目的:探讨乳腺癌组织中survivin基因和c-erbB-2基因的表达与预后的关系。方法:用免疫组化S-P法检测80例乳腺癌组织中survivin、c-erbB-2的表达,分析其与腋淋巴结转移和5年无病生存率之间的关系。结果:survivin基因在乳腺癌中阳性表达率为68.75%(55/80)、c-erbB-2基因阳性表达率为36.25%(29/80),均与腋淋巴结转移正相关,与5年生存率负相关(P<0.05);survivin和c-erbB-2表达与肿瘤病理类型、发病年龄、临床分期无明显相关性(P>0.05);survivin和c-erbB-2基因表达相互呈正相关(P<0.05)。结论:凋亡抑制基因survivin和原癌基因c-erbB-2可能在乳腺癌的发生、发展过程中起重要作用,联合检测能更好地判断乳腺癌的预后。     

Expression of ER, PgR, HER1, HER2, and response: a study of preoperative chemotherapy.

PURPOSE: To identify the role of estrogen (ER), progesterone (PgR), epidermal growth factor 1 (HER1), and HER2 receptors in predicting response to preoperative chemotherapy. MATERIALS AND METHODS: We reviewed the pretreatment biopsies of 485 patients with locally advanced breast cancer (cT2-T4, N0-2, M0) treated with preoperative chemotherapy. The incidence of pathological complete remission (pCR) and outcome were assessed with respect to clinical and pathological findings including ER/PgR status (absent versus expressed), HER1 (absent versus expressed) and HER2 (overexpressed versus none) expression. RESULTS: Patients with ER/PgR-absent tumors were 12.0 times [95% confidence interval (CI) 4.93-29.28] more likely to achieve a pCR (P < 0.0001). Predictors of disease-free survival (DFS) at the univariate analysis included HER1 [hazards ratio (HR) 1.6, 95% CI 1.04-2.32, P = 0.03] and HER2 (HR 1.6, 95% CI 1.08-2.38, P = 0.02) expression. A statistically significant difference in DFS was confirmed at the multivariate analysis for patients with ER/PgR-absent disease (HR 2.1, 95% CI 1.41-2.99, P = 0.0002). CONCLUSIONS: The pCR rate is higher and outcome worse for patients with ER/PgR-absent tumors. HER1 and HER2 expression may have a prognostic role in locally advanced breast cancer and warrant further studies.     

Tumor tissue levels of tissue inhibitor of metalloproteinase-1 as a prognostic marker in primary breast cancer.

PURPOSE: In the present study, we investigated the association between tumor tissue levels of tissue inhibitor of metalloproteinase-1 (TIMP-1) and prognosis in patients with primary breast cancer and analyzed whether TIMP-1 may be useful as a prognostic marker in combination with urokinase plasminogen activator (uPA) and plasminogen activator inhibitor type-1 (PAI-1). EXPERIMENTAL DESIGN: In cytosolic extracts of 2984 primary breast tumors, total levels of TIMP-1 were determined using an established, validated ELISA. Levels of uPA and PAI-1 have previously been determined in the extracts. RESULTS: Univariate survival analysis showed a significant relationship between higher levels of TIMP-1 (continuous log-transformed variable) and poor prognosis [recurrence-free survival (RFS), overall survival (OS); P < 0.001]. Performing isotonic regression analysis, we identified a cut point to classify tumors as TIMP-1-low or TIMP-1-high. Using this cut point, high levels of TIMP-1 were significantly associated with shorter survival in univariate analysis, both in the total patient group (RFS, OS; P < 0.001), in the node-negative subgroup (RFS, hazard ratio = 1.28, P = 0.006), and in the node-positive subgroup (RFS, hazard ratio = 1.43, P < 0.001). In multivariate analysis, including uPA and PAI-1, TIMP-1 was significantly associated with shorter RFS, both when included as a continuous log-transformed (P = 0.03) and as a dichotomized variable (P = 0.002). CONCLUSIONS: This study validates previous findings that tumor tissue levels of TIMP-1 are associated with prognosis in patients with primary breast cancer. It confirms that TIMP-1 may be useful as a prognostic marker in combination with uPA/PAI-1 and adds substantial positive information on the use of TIMP-1 as a prognostic marker in breast cancer.     

Dissemination risk index based on plasminogen activator system components in primary breast cancer.

PURPOSE: To study interactions between disease-free survival (DFS) and four components of the plasminogen activator system: urokinase-type plasminogen activator (uPA), its two inhibitors (PAI-1 and PAI-2), and its membrane receptor uPAR. PATIENTS AND METHODS: We conducted a retrospective study of 499 primary breast cancer patients (median follow-up, 6 years). uPA, PAI-1, and PAI-2 were determined on cytosols and uPAR on solubilized pellets, using enzyme-linked immunoadsorbent assay kits (American Diagnostica, Greenwich, CT). Classical univariate and multivariate statistical methods were used together with multiple correspondence analysis to graphically examine interactions between the variables and outcome. RESULTS: By univariate analysis, higher uPA and PAI-1 values were significantly related to shorter DFS (P =.002; P <.00002). PAI-2 was not significantly related to DFS, although patients with high and very low PAI-2 values had a longer DFS. Multiple correspondence analysis showed the parallel impact of uPA and PAI-1 on outcome, and the clearly different behavior of PAI-2 compared with PAI-1. The prognostic contribution of uPAR seemed weak by both methods. A dissemination risk index [uPA x PAI-1/(PAI-2 + 1)], taking into account the modulation of uPA proteolytic activity by the ratio of its two inhibitors, was then tested. Dissemination risk index was selected as an independent variable in the Cox model in the overall population (P <.000001) and in node-positive patients (P <.00001). It was the only variable selected in node-negative patients (P =. 003). CONCLUSION: A dissemination risk index determined on primary tumor and taking into account the different effects of PAI-1 and PAI-2 on uPA can be of major help in clinical management of breast cancer, particularly in node-negative patients.