Prognostical value of classical and molecular biomarkers in the follow-up of early breast cancer

Uvod: Novi molekularni biomarkeri (uPA, PAI-1, katepsin D), uz klasične prognostičke markere, mogu biti korisni u prepoznavanju visoko-rizične podgrupe pacijenata u relativno dobroj prognostičkoj grupi pacijenata sa karcinomom dojke kod kojih nisu zahvaćeni limfni čvorovi (pN0). Pacijenti i metode: Procenjivana je prognostička vrednost urokinaznog plazminog aktivatora (uPA), plazminogen aktivator inhibitora-1 (PAI-1) i katepsina D u grupi postmenopauznih pacijentkinja operisanih zbog karcinoma dojke, kod kojih tumori pokazuju pozitivan steroidni receptorski status (estrogen receptor (ER)/progesteron receptor (PR), a kod kojih nije primenjivana adjuvantna terapija. Značaj biomarkera je analiziran tokom tri različita perioda praćenja. Analizirana su 73 uzorka citosolnog ekstrakta. Vrednosti ER i PR su odreĎivane klasičnom biohemijskom metodom, katepsin D radiometrijskim imuno-esejom a uPA i PAI-1 ELISA testom. Vrednosti HER 2 genske amplifikacije su odreĎivane hromogenom in situ hibridizacijom (CISH). Rezultati: Tokom ranog perioda praćenja (prvih pet godina nakon hirurškog lečenja) pacijenti sa veličinom tumora pT1 i niskim vrednostima PAI-1 (PAI-1<6.35 pg/mg) su imali bolju prognozu u odnosu na pacijente sa veličinom tumora (pT2,3) ili koji su imali visoke vrednosti PAI-1. Analizom 4 fenotipa definisanih veličinom tumora i PAI-1 statusom pokazano je da pacijenti koji imaju tumore veličine pT1, bez obzira na nivo PAI-1, ili koji imaju tumore veličine pT2,3 uz niske vrednosti PAI-1 imaju sličan DFI i dobru prognozu kao i pacijenti sa veličinom tumora pT2,3 uz visoke vrednosti PAI-1. Zakljuĉak: Veličina tumora u kombinaciji sa PAI-1 ima prognostički i moguće prediktivni značaj tokom ranog praćenja postmenopauznih pacijentkinja sa karcinomom dojke kod kojih tumori pokazuju pozitivan steroidni receptorski status (ER/PR), histološkog gradusa II.Introduction: New molecular prognostic biomarkers (uPA, PAI-1, cathepsine D), together with classical prognostic markers, might be useful to select high risk subgroup of patients within relatively good prognostic group of breast carcinoma patients without metastases in lymph nodes (pN0). Patients and methods: We evaluated urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor-1 (PAI-1) prognostic value in postmenopausal, node-negative breast cancer patients bearing tumors with estrogen receptor (ER)/progesterone receptor (PR) expression, treated with locoregional therapy alone, within 3 different follow-up periods: first five years after surgical treatment, second five years and during the who period of ten years. We focused our analysis on tumors of histological grade II in order to improve its prognostic value and, consequently, to improve a decision making process. The cytosol extracts of 73 tumor samples were used for assessing several biomarkers. ER and PR levels were measured by classical biochemical method. Cathepsin D was assayed by a radiometric immunoassay while both uPA and PAI-1 level determinations were performed by enzyme-linked immunosorbent assays. HER-2 gene amplification was determined by chromogenic in situ hybridization (CISH) in primary tumor tissue. Results: During the early follow up patients bearing tumors smaller than or equal to 2 cm (pT1) or those with low PAI-1 levels (PAI-1<6.35 pg/mg) showed favorable outcome compared to patients bearing tumors greater than 2 cm (pT2,3) or those with high PAI-1 levels, respectively. Analyses of 4 phenotypes, defined by tumor size and PAI-1 status, revealed that patients bearing either pT1 tumors, irrespective of PAI-1 levels, or pT2,3 tumors with low PAI-1 levels, had similar disease-free interval probabilities and showed favorable outcome compared to those bearing pT2,3 tumors with high PAI-1 levels.Conclusion: Our findings suggest that tumor size and PAI-1, used in combination as phenotypes are not only prognostic but might also be predictive in node-negative, postmenopausal breast cancer patients bearing histological grade II tumors with ER/PR expression, during an early follow-up period

Intraductal papilloma of ectopic breast tissue in axillary lymph node of a patient with a previous intraductal papilloma of ipsilateral breast: a case report and review of the literature

The presence of ectopic breast tissue in axillary lymph nodes (ALN) is a benign condition that must be differentiated from primary or metastatic carcinoma. Here we report a patient who underwent excision of enlarged ALN 10 years after she had received surgical treatment of ipsilateral breast for an intracystic intraductal papilloma (IDP). Histological examination of the removed ALN revealed that the proliferative lesion consisted of papillary and tubular structures lined by luminal cuboidal cells and a distinct outer layer of myoepithelial cells resembling IDP of the breast. Immunostaining with a set of immunohistochemical markers including AE/AE3, alpha-smooth muscle actin and p63 in combination with estrogen and progesterone receptors confirmed the diagnosis of ectopic IDP

Trefoil Factor 1 in Early Breast Carcinoma: A Potential Indicator of Clinical Outcome during the First 3 Years of Follow-Up

Background. A role of an estrogen-regulated, autocrine motogenic factor was assumed to be a major biological role of trefoil factor 1 (TFF1) in breast cancer. TFF1 is regarded as a predictive factor for positive response to endocrine therapy in breast cancer patients. The aim of our study was to examine TFF1 level distribution in breast carcinomas in order to distinguish estrogen-independent from estrogen-dependent TFF1 expression and to evaluate clinical usefulness of TFF1 status in early breast cancer during the first 3 years of follow-up. Methods. The study included 226 patients with primary operable invasive early breast carcinomas for whom an equal, a 3-year follow-up was conducted. TFF1 levels as well as estrogen receptor (ER) and progesterone receptor (PR) levels were measured in cytosolic extracts of tumor samples by immunoradiometric assay or by use of classical biochemical method, respectively. Non-parametric statistical tests were applied for data analyses. Results. Statistical analysis revealed that TFF1 levels were significantly higher in premenopausal patients (p=0.02), or in tumors with: lower histological grade (p LT 0.001), positive ER or PR status (p LT 0.001, in both cases). On the basis of TFF1 level distribution between ER-negative and ER-positive postmenopausal patients with tumors of different histological grade, 14 ng/mg was set as the cut-off value to distinguish estrogen-independent from estrogen-dependent TFF1 expression in breast cancer. Depending on menopausal and PR status, positive TFF1 status identified patients at opposite risk for relapse among ER-positive patients with grade II tumors. Among ER-and and PR-positive premenopausal patients with grade II tumors, TFF1 status alone identified patients at opposite risk for relapse. Conclusions. Determination of TFF1 status might identify patients at different risk for relapse and help in making decision on administering adjuvant therapy for early breast cancer patients during the first 3 years of follow-up

Angiogenesis: bFGF and VEGF in breast carcinoma

Angiogenesis, or neovascularization, is a complex process leading to formation of new blood vessels from the pre-existing vascular network of the tissue. Angiogenesis plays a central role in various physiological and pathological conditions, including embryonic development, reproduction, inflammation and wound healing, infertility, heart diseases, ulcers, rheumatoid arthritis, diabetic blindness and cancer. It is a multistep process involving EC activation, basement membrane and extracellular matrix (ECM) degradation, EC proliferation, migration and differentiation, synthesis of new basement membrane and maturation of new blood vessels. Tumor vasculature is considered to be of an "immature" nature with series of structural abnormalities. There are reciprocal paracrine interactions between ECs, tumor cells, stroma and ECM. Angiogenesis plays a key role in transformation of normal to malignant cell, tumor progression and metastasis. It is similar to the metastatic process in that it requires EC attachment, proteolysis, and locomotion to proceed. A close relationship exists between the tumor and ECs invasiveness of the tissue. The switch to the angiogenic phenotype involves a change in the local equilibrium between positive and negative regulators of the growth of microvessels. Basic fibroblast growth factor (bFGF) and vas­cular endothelial growth factor (VEGF) are positive regulators of angiogenesis. Intimate cross-talk exists among bFGF and the different members of the VEGF family during angiogenesis, lymphangiogenesis, and vasculogenesis. A substantial body of experimental evidence supports the hypothesis that angiogenesis and angiogenic factors may be strong prognostic and predictive factors in breast carcinoma. This article reviews the current knowledge on angiogenesis and its positive regulators: bFGF and VEGF.

Cathepsin D as an indicator of clinical outcome in early breast carcinoma during the first 3 years of follow-up

Aim: The aim of this study was to evaluate clinical usefulness of cathepsin D status in early breast cancer during the first 3 years of follow-up. Patients and methods: The study included 226 patients with histologically verified, primary operable invasive early breast carcinomas. Concentrations of estrogen receptor (ER) and progesterone receptor (PR) in breast tumor cytosols were measured by use of the classical biochemical method. The concentration of three cathepsin D forms (52-, 48- and 34-kDa proteins) was determined by a radioimmunoassay Results: On the basis of differences in cathepsin D levels either within an ER-/PR- phenotype or between this and either ER+/PR+ or ER+/PR- phenotypes, a concentration of 39 pmol/mg was determined as the cutoff value for distinguishing estrogen-regulated cathepsin D expression. Estrogen-regulated cathepsin D expression was recognized as a high-risk biomarker for low-risk (histological grade I) breast cancer patients and as a low-risk biomarker for high-risk patients (pN(+) pT2,3). Conclusion: Determination of cathepsin D status in breast cancer might identify patients at different risk for relapse and might facilitate the selection of more or less aggressive adjuvant therapy for early breast cancer patients during the first 3 years of follow-up