BACKGROUND: Urokinase-type plasminogen activator (uPA) and its inhibitor
 (PAI-1) play essential roles in tumor invasion and metastasis. High levels
 of both uPA and PAI-1 are associated with poor prognosis in breast cancer
 patients. To confirm the prognostic value of uPA and PAI-1 in primary
 breast cancer, we reanalyzed individual patient data provided by members
 of the European Organization for Research and Treatment of Cancer-Receptor
 and Biomarker Group (EORTC-RBG). METHODS: The study included 18 datasets
 involving 8377 breast cancer patients. During follow-up (median 79
 months), 35% of the patients relapsed and 27% died. Levels of uPA and
 PAI-1 in tumor tissue extracts were determined by different immunoassays;
 values were ranked within each dataset and divided by the number of
 patients in that dataset to produce fractional ranks that could be
 compared directly across datasets. Associations of ranks of uPA and PAI-1
 levels with relapse-free survival (RFS) and overall survival (OS) were
 analyzed by Cox multivariable regression analysis stratified by dataset,
 including the following traditional prognostic variables: age, menopausal
 status, lymph node status, tumor size, histologic grade, and steroid
 hormone-receptor status. All P values were two-sided. RESULTS: Apart from
 lymph node status, high levels of uPA and PAI-1 were the strongest
 predictors of both poor RFS and poor OS in the analyses of all patients.
 Moreover, in both lymph node-positive and lymph node-negative patients,
 higher uPA and PAI-1 values were independently associated with poor RFS
 and poor OS. For (untreated) lymph node-negative patients in particular,
 uPA and PAI-1 included together showed strong prognostic ability (all
 P<.001). CONCLUSIONS: This pooled analysis of the EORTC-RBG datasets
 confirmed the strong and independent prognostic value of uPA and PAI-1 in
 primary breast cancer. For patients with lymph node-negative breast
 cancer, uPA and PAI-1 measurements in primary tumors may be especially
 useful for designing individualized treatment strategies.

/pdf/pooled-analysis-of-prognostic-impact-of-urokinase-type-wfsddi9we3.pdf

Pooled analysis of prognostic impact of urokinase-type plasminogen activator and its inhibitor PAI-1 in 8377 breast cancer patients.

5-fluorouracil and other fluoropyrimidines in colorectal cancer: Past, present and future

Proliferative markers as prognostic and predictive tools in early breast cancer: where are we now?

Cancer is a leading cause of death worldwide and accounts for several millions of deaths every year. The survival of cancer patients is strongly associated with the stage of the tumor at the time of diagnosis. Identifying the cancer at the cellular level in an early stage before metastasis holds great promise for increasing the survival of cancer patients. A major focus of research towards this goal is on the estimation of abnormalities in gene expression in living cells. Tumor-related mRNA has been widely used as a specific marker to assess the migration of tumor cells locally or in the bloodstream. Changes in the level of tumor-related mRNA expression are correlated with tumor burden and malignant progression. The detection of tumor-related mRNA markers in intact cancer cells provides new tools for identifying cancer cells in clinical samples. Recently, a variety of techniques have been exploited to monitor tumor-related mRNAs. Among these methods, fluorescence imaging analysis offers an appealing approach for the detection of cancer at the cellular level, which may be of prognostic significance. Many fluorescence probes have been synthesized for the detection and imaging of mRNA in cancer cells and most of the research focuses on detecting a single type of mRNA, which may yield false positive results and limits the development of intracellular mRNA imaging and detection. Notably, cancer is associated with multiple tumor-related mRNAs, and some mRNA markers are expressed in normal cells. Simultaneous detection of multiple targets brings new opportunities for improving the accuracy of early cancer detection over the single-marker assay. Although various types of platforms for the detection of multiple targets have been developed, none have been designed for imaging three or more markers in living cells. Such intracellular imaging techniques for multiple tumorrelated mRNAs could promote the progress of early cancer detection. Herein, we describe a multicolor fluorescence nanoprobe based on nanoflares, which simultaneously detects three intracellular tumor-related mRNAs. The nanoprobe consists of gold nanoparticles (Au NPs) functionalized with a dense shell of recognition sequences (synthetic oligonucleotides) hybridized to three short dye-terminated reporter sequences by gold–thiol bond formation (Figure 1). The recognition sequences contain 21-base recognition elements for three specific mRNA transcripts: c-myc mRNA, TK1 mRNA, and GalNAc-T mRNA.

A multicolor nanoprobe for detection and imaging of tumor-related mRNAs in living cells.

A new class of intracellular nanoprobe, termed fluorescence resonance energy transfer (FRET) nanoflares, was developed to sense mRNA in living cells. It consists of a gold nanoparticle (AuNP), recognition sequences, and flares. Briefly, the AuNP functionalized with recognition sequences hybridized to flares, which are designed as hairpin structures and fluorescently labeled donors and acceptors at two ends, respectively. In the absence of targets, the flares are captured by binding with the recognition sequences, separating of the donor and acceptor, and inducing low FRET efficiency. However, in the presence of targets, the flares are gradually displaced from the recognition sequences by the targets, subsequently forming hairpin structures that bring the donor and acceptor into close proximity and result in high FRET efficiency. Compared to the conventional single-dye nanoflares, the upgraded FRET nanoflares can avoid false positive signals by chemical interferences (such as nuclease and GSH) and thermodynamic fluctuations. Moreover, the signal generation in FRET nanoflares can be easily made with ratiometric measurement, minimizing the effect of system fluctuations.

FRET Nanoflares for Intracellular mRNA Detection: Avoiding False Positive Signals and Minimizing Effects of System Fluctuations

PURPOSE: To assess the prognostic value of thymidine kinase (TK), an enzyme involved in the DNA synthesis salvage pathway, relative to other prognostic factors in primary breast cancer. PATIENTS AND METHODS: This retrospective study involved 1,692 patients with operable breast cancer treated in six institutions (median follow-up, 82 months). Among the 857 node-negative patients, 135 received adjuvant chemotherapy (fluorouracil, doxorubicin, cyclophosphamide [FAC] or fluorouracil, etoposide, and cisplatin [FEC]). TK was assayed in cytosol with a quantitative radioenzymatic technique. Disease-specific survival (DSS), local recurrence-free interval (LRI), and distant-relapse-free interval (DRI) were investigated. RESULTS: High TK levels were associated with large tumor size, high histologic grade, and steroid hormone receptor negativity. Univariate analysis of the entire data set showed that high TK levels were related to shorter DSS (P < 10-5), LRI (P < 10-3), and DRI (P < 10-5). In time-dependent Cox model...

Thymidine Kinase as a Proliferative Marker: Clinical Relevance in 1,692 Primary Breast Cancer Patients

The purpose of this retrospective multicentre study was to assess the prognostic value of urokinase plasminogen activator (uPA) and p53 levels in a large series of primary breast cancer, using an automatic quantitative luminometric method. Samples of 1245 operable breast tumours were collected from seven French institutions and patients were followed for a median of 75 months. The median uPA and p53 levels assayed in cytosols by means of the immunoluminometric technique (LIA) were 0.31 and 0.20 ng mg(-1) of protein respectively. In univariate analysis, high levels of uPA and p53 were associated with shorter disease-specific survival, disease-free interval, and distant recurrence-free interval. The 5-year survival rates were 95.5% among patients with uPA values below the 20th percentile, and 77.5% in those with values above the 80th percentile. The 5-year survival rates were 91.0% in patients with p53 values below the 20th percentile, and 77.6% in those with values above the 80th percentile. In multivariate analysis, the risk of disease-related death increased with uPA levels after adjustment for tumour size, histological grade, lymph node involvement, and estrogen receptor status. A high level of uPA was also related to a shorter disease-free interval and distant recurrence-free interval. In node-negative patients, a high level of uPA remained strongly related to the three outcomes. When adjusted for other prognostic factors, p53 was no longer significantly related to the outcomes. Given its rapidity and simple application to routinely prepared cytosols, this LIA may be useful for evaluating the prognostic impact of uPA in primary breast cancer, particularly in node-negative patients. According to our results, the prognostic value of p53 accumulation is limited when uPA is included in multivariate analysis.

/pdf/prognostic-value-of-upa-and-p53-accumulation-measured-by-11qfk42yfc.pdf

G Ricolleau

Papers

Thymidine Kinase as a Proliferative Marker: Clinical Relevance in 1,692 Primary Breast Cancer Patients

Prognostic value of uPA and p53 accumulation measured by quantitative biochemical assays in 1245 primary breast cancer patients: a multicentre study