MMP9 but Not EGFR, MET, ERCC1, P16, and P-53 Is Associated with Response to Concomitant Radiotherapy, Cetuximab, and Weekly Cisplatin in Patients with Locally Advanced Head and Neck Cancer

Concomitant administration of radiotherapy with cisplatin or radiotherapy with cetuximab appear to be the treatment of choice for patients with locally advanced head and neck cancer. In the present retrospective analysis, we investigated the predictive role of several biomarkers in an unselected cohort of patients treated with concomitant radiotherapy, weekly cisplatin, and cetuximab (CCRT). We identified 37 patients treated with this approach, of which 13 (35%) achieved a complete response and 10 (27%) achieved a partial response. Severe side effects were mainly leucopenia, dysphagia, rash, and anemia. Tumor EGFR, MET, ERCC1, and p-53 protein and/or gene expression were not associated with treatment response. In contrast, high MMP9 mRNA expression was found to be significantly associated with objective response. In conclusion, CCRT is feasible and active. MMP9 was the only biomarker tested that appears to be of predictive value in cetuximab treated patients. However, this is a hypothesis generating study and the results should not be viewed as definitive evidence until they are validated in a larger cohort

Quantitative determination of steroid hormones and their metabolites in the systemic circulation of women with benign and malignant breast diseases and healthy individuals, with current analytical techniques

Steroid hormones affect major biological functions and play a critical role in breast carcinogenesis. They are potent enough so as to exert significant effects even at low concentrations, while level variations can be used for diagnostic purposes. Quantitative determination of steroid hormones by liquid chromatography-mass spectrometry raises new perspectives in the study of hormone-dependent malignancies. The goal of this study was 2-fold: first to develop and validate an LC-MS method for the simultaneous quantification of androsterone glucuronide (ADTG), androstane-3α, 17β-diol-17glucuronide (3α-diol-17G), dehydroepiandrosterone sulfate (DHEAS), androstenedione (A4) and estrone sulfate (E1S), and second and equally important, to apply the method in the study of breast cancer. A simple and efficient fit-for-purpose method for the simultaneous quantification of the five hormones in human plasma was developed and validated. The presented method permits omission of derivatization and requires a single solid-phase extraction (SPE) procedure while the chromatographic separation can be achieved on a single C18 analytical column for all five analytes. The validated method was successfully applied for the analysis of 191 human plasma samples from postmenopausal women, which were divided into four groups according to the diagnosis of the breast lesion: benign breast disease (BBD), lobular neoplasia (LN), ductal carcinoma in situ (DCIS) and invasive ductal carcinoma (IDC).To the best of our knowledge, this is the first multi-analyte study evaluating the levels of all five hormones in patients with different histological types of breast diseases, which reflect different evolutionary stages of breast cancer. Significantly higher DHEAS plasma levels were observed in LN patients compared to IDC and BBD patients (p<0.05). LN patients also exhibited higher levels of DHEAS compared to DCIS patients, nevertheless the difference was marginally significant (p=0.057). Additionally, ADTG levels were significantly higher in patients with LN compared to those with BBD (p<0.05). Potential correlations of ADTG, 3a-diol-17G, DHEAS, A4 and E1S levels with conventional tumor markers (CEA, CA 15-3, Her/2-neu) levels and with anthropometric and epidemiologic characteristics of the patients, were also investigated.The results of the study are of particular interest for lobular neoplasia as it is recognized as a non-obligate precursor of invasive breast cancer and as a marker of increased risk for subsequent breast carcinoma development. Lobular neoplastic lesions are clinically occult and often mammographically silent. Thus, their differential diagnosis from other breast lesions is of crucial importance for the monitoring of patients and for selecting the appropriate treatment. The findings of the study support the potential application of ADTG and DHEAS quantitative analysis by LC-MS in the differential diagnosis of breast diseases. Prospectively, these two steroid hormones could be considered for inclusion in a breast cancer risk prediction model. Their inclusion could reinforce the valuable contribution of these models in the identification of women, which are in a greater risk and in need of a through diagnostic control and of intense systematic monitoring.Οι στεροειδείς ορμόνες επηρεάζουν σημαντικές βιολογικές λειτουργίες και παίζουν καίριο ρόλο στην καρκινογένεση στο μαστό. Πρόκειται για ισχυρά μόρια, τα οποία ασκούν σημαντικές επιδράσεις ακόμα και σε χαμηλές συγκεντρώσεις, ενώ οι μεταβολές στα επίπεδα τους μπορούν να χρησιμοποιηθούν για διαγνωστικούς σκοπούς. Ο ποσοτικός προσδιορισμός των στεροειδών ορμονών με υγρή χρωματογραφία-φασματομετρία μαζών (LC-MS) δημιουργεί νέες προοπτικές στη μελέτη των ορμονοευαίσθητων κακοηθειών. Ο σκοπός αυτής της μελέτης ήταν διττός. Πρώτον, η ανάπτυξη και επικύρωση μεθόδου LC-MS για την ταυτόγχρονη ποσοτικοποίηση του γλυκουρονιδίου της ανδροστερόνης (ADTG), του ανδροσταν-3α, 17β-διολ-17-γλυκουρονιδίου (3α-diol-17G), της θειϊκής δεϋδροεπιανδροστερόνης (DHEAS), της ανδροστενδιόνης (A4) και της θειϊκής οιστρόνης (E1S). Δεύτερον και εξίσου σημαντικό, ήταν η εφαρμογή της αναπτυχθείσας μεθόδου στη μελέτη του καρκίνου του μαστού. Πραγματοποιήθηκε ανάπτυξη και επικύρωση μιας απλής και αποδοτικής μεθόδου LC-MS για τον ταυτόχρονο ποσοτικό προσδιορισμό και των πέντε ορμονών σε ένα δείγμα πλάσματος. Η μέθοδος περιλαμβάνει μια ενιαία κατεργασία δείγματος, για το σύνολο των ορμονών, με απαλοιφή της παραγωγοποίησης, ενώ ο χρωματογραφικός διαχωρισμός πραγματοποιείται σε μια C18 αναλυτική στήλη. Η μέθοδος εφαρμόστηκε με επιτυχία στην ανάλυση 191 δειγμάτων πλάσματος μετεμμηνοπαυσιακών γυναικών, οι οποίες καταχωρήθηκαν σύμφωνα με τη διάγνωση σε τέσσερις ομάδες: καλοήθη νόσο του μαστού (BBD), λοβιακή νεοπλασία (LN), πορογενές καρκίνωμα in situ (DCIS) και διηθητικό πορογενές καρκίνωμα (IDC). Η παρούσα, είναι η πρώτη μελέτη συνδυαστικής ανάλυσης και σύγκρισης των επιπέδων των πέντε ορμονών σε ασθενείς με παθήσεις του μαστού διαφορετικού ιστολογικού τύπου, οι οποίες αντικατοπτρίζουν διαφορετικά εξελικτικά στάδια του καρκίνου του μαστού. Σημαντικά υψηλότερα επίπεδα DHEAS παρατηρήθηκαν στο πλάσμα των ασθενών με LN έναντι των ασθενών με IDC και BBD (p<0,05). Οι ασθενείς με LN εμφάνισαν υψηλότερα επίπεδα DHEAS και έναντι των ασθενών με DCIS, αν και η διαφορά ήταν οριακά στατιστικά σημαντική (p=0,057). Επιπρόσθετα, τα επίπεδα του ADTG βρέθηκαν να είναι σημαντικά υψηλότερα στις ασθενείς με LN σε σύγκριση με τις γυναίκες με BBD (p<0,05). Διερευνήθηκε επίσης η συσχέτιση μεταξύ των επιπέδων των ADTG, 3α-diol-17G, DHEAS, A4 και E1S με τα επίπεδα καθιερωμένων καρκινικών δεικτών (CEA, CA 15-3, Her/2-neu) καθώς και με επιλεγμένα ανθρωπομετρικά και επιδημιολογικά στοιχεία των ασθενών. Τα αποτελέσματα της μελέτης παρουσιάζουν ιδιαίτερο ενδιαφέρον για τη λοβιακή νεοπλασία, η οποία θεωρείται εκτός από πρόδρομη αλλοίωση και δείκτης αυξημένου κινδύνου για επακόλουθη ανάπτυξη καρκίνου στο μαστό. Οι LN αλλοιώσεις είναι κλινικά απόκρυφες και συνήθως μαστογραφικά σιωπηλές. Η διαφοροδιάγνωση τους είναι εξαιρετικά σημαντική όχι μόνον για την παρακολούθηση των ασθενών αλλά και για τη χορήγηση κατάλληλης θεραπείας. Τα ευρήματα της μελέτης υποστηρίζουν ότι η ποσοτική ανάλυση των ADTG και DHEAS με την εφαρμογή της LC-MS, μπορεί να αποτελέσει ένα ωφέλιμο εργαλείο στη διαφορική διάγνωση των παθήσεων του μαστού. Προοπτικά, οι δυο αυτές ορμόνες θα μπορούσαν να συμπεριληφθούν σε ένα μοντέλο πρόβλεψης εμφάνισης της νόσου. Η ένταξη τους, θα μπορούσε να ενισχύσει την πολύτιμη συνεισφορά των μοντέλων στην ταυτοποίηση γυναικών, οι οποίες ευρίσκονται σε μεγαλύτερο κίνδυνο και χρήζουν πληρέστερου διαγνωστικού ελέγχου και συστηματικής παρακολούθησης

Circulating levels of matrix metalloproteinase-9 (MMP-9), neutrophil gelatinase-associated lipocalin (NGAL) and their complex MMP-9/NGAL in breast cancer disease

<p>Abstract</p> <p>Background</p> <p>Recent evidence suggests that neutrophil gelatinase-associated lipocalin (NGAL) expression is induced in many types of human cancer, while detection of its complex with matrix metalloproteinase-9 (MMP-9) is correlated with cancer disease status. We aim to evaluate the serum expression of MMP-9, NGAL and their complex (MMP-9/NGAL) during the diagnostic work-up of women with breast abnormalities and investigate their correlation with disease severity.</p> <p>Methods</p> <p>The study included 113 women with non-palpable breast lesions undergoing vacuum-assisted breast biopsy for histological diagnosis, and 30 healthy women, which served as controls. Expression levels of MMP-9, NGAL and their complex MMP-9/NGAL were determined in peripheral blood samples with immunoenzymatic assays.</p> <p>Results</p> <p>Women with invasive ductal carcinoma exhibited significantly increased levels of MMP-9, NGAL and MMP-9/NGAL compared to healthy controls (MMP-9: p < 0.003, NGAL: p < 0.008 MMP-9/NGAL: p < 0.01). Significant correlations were observed between MMP-9 and NGAL serum levels and breast disease severity score (r = 0.229, p < 0.006 and r = 0.206, p < 0.01, respectively), whereas a non-significant correlation was found for their complex. MMP-9, NGAL and their complex MMP-9/NGAL levels were not correlated with either Body Mass Index (BMI) or age of patients.</p> <p>Conclusion</p> <p>These findings suggest that the serum measurement of MMP-9 and NGAL may be useful in non-invasively monitoring breast cancer progression, while supporting their potential role as early biomarkers of breast disease status.</p

Decreased circulating levels of angiopoietin – 1 (Ang-1) are associated with the presence of multinodular goiter or differentiated thyroid cancer

Background: The aim of this study was to investigate whether the presence of benign or malignant nodular thyroid disease affects levels of circulating angiogenesis cytokines. Methods: In this study we investigated levels of angiopoietin – 1 and -2 (Ang-1 and Ang-2 respectively), vascular endothelial growth factor –A (VEGF-A), galectin-3 (Gal-3), urokinase plasminogen activator receptor (uPAR) and plasminogen activation inhibitor – 1 (PAI-1) in 40 patients with differentiated thyroid cancer (DTC), 45 with thyroid papillary microcarcinoma (mPTC), 53 patients with multinodular goiter (MNG) and 58 controls. Six months after surgery 28 patients resubmitted blood samples. The diagnostic value of Ang-1 levels was evaluated with receiver operating characteristic (ROC) curves. Results: Statistically significant lower levels of Ang-1 were observed in DTC and MNG patients compared to controls (p<.05). No significant differences were observed in the levels of the other factors. The area under ROC curves for Ang-1 discerning DTC, mPTC and MNG from control were 0.68, 0.66 and 0.71 respectively. A significant increase in Ang-1 levels (p<.05) was documented in the subset of patients that underwent thyroidectomy. Thyroidectomy did not influence levels of the other factors. Conclusions: Our results suggest an association between low levels of Ang-1 and the presence of underlying benign or malignant nodular thyroid disease

Bioanalytical LC–MS Method for the Quantification of Plasma Androgens and Androgen Glucuronides in Breast Cancer

The physiological and pathological development of the breast is strongly affected by the hormonal milieu consisting of steroid hormones. Mass spectrometry (MS) technologies of high sensitivity and specificity enable the quantification of androgens and consequently the characterization of the hormonal status. The aim of this study is the assessment of plasma androgens and androgen glucuronides, in the par excellence hormone-sensitive tissue of the breast, through the application of liquid chromatography–mass spectrometry (LC–MS). A simple and efficient fit-for-purpose method for the simultaneous identification and quantification of dehydroepiandrosterone sulfate (DHEAS), androstenedione (A4), androsterone glucuronide (ADTG) and androstane-3α, 17β-diol-17-glucuronide (3α-diol-17G) in human plasma was developed and validated. The presented method permits omission of derivatization, requires a single solid-phase extraction procedure and the chromatographic separation can be achieved on a single C18 analytical column, for all four analytes. The validated method was successfully applied for the analysis of 191 human plasma samples from postmenopausal women with benign breast disease (BBD), lobular neoplasia (LN), ductal carcinoma in situ and invasive ductal carcinoma (IDC). DHEAS plasma levels exhibited significant differences between LN, IDC and BBD patients (P < 0.05). Additionally, ADTG levels were significantly higher in patients with LN compared with those with BBD (P < 0.05)

Bioanalytical LC-MS Method for the Quantification of Plasma Androgens and Androgen Glucuronides in Breast Cancer

The physiological and pathological development of the breast is strongly affected by the hormonal milieu consisting of steroid hormones. Mass spectrometry (MS) technologies of high sensitivity and specificity enable the quantification of androgens and consequently the characterization of the hormonal status. The aim of this study is the assessment of plasma androgens and androgen glucuronides, in the par excellence hormone-sensitive tissue of the breast, through the application of liquid chromatography-mass spectrometry (LC-MS). A simple and efficient fit-for-purpose method for the simultaneous identification and quantification of dehydroepiandrosterone sulfate (DHEAS), androstenedione (A4), androsterone glucuronide (ADTG) and androstane-3α, 17β-diol-17-glucuronide (3α-diol-17G) in human plasma was developed and validated. The presented method permits omission of derivatization, requires a single solid-phase extraction procedure and the chromatographic separation can be achieved on a single C18 analytical column, for all four analytes. The validated method was successfully applied for the analysis of 191 human plasma samples from postmenopausal women with benign breast disease (BBD), lobular neoplasia (LN), ductal carcinoma in situ and invasive ductal carcinoma (IDC). DHEAS plasma levels exhibited significant differences between LN, IDC and BBD patients (P < 0.05). Additionally, ADTG levels were significantly higher in patients with LN compared with those with BBD (P < 0.05)

Serum Concentration of Selected Angiogenesis-Related Molecules Differs among Molecular Subtypes, Body Mass Index and Menopausal Status in Breast Cancer Patients

Background: Angiogenesis is a hallmark of breast cancer (BC) and is mediated by the vascular endothelial growth factor (VEGF) signaling axis. It is regulated by different proangiogenic factors, including platelet-derived growth factor-CC (PDGF-CC) and heparin-binding EGF-like growth factor (HB-EGF), as well as co-receptors, such as neuropilin-1, which could have prognostic implications in BC patients. Patients and methods: We assessed the serum levels of VEGF, HB-EGF, PDGF-CC and neuropilin-1 in 205 patients with early BC (invasive, n = 187; in situ, n = 18) and in 31 healthy donors (HD) and investigated the potential associations with clinical and histopathological parameters. Results: VEGF serum levels were significantly higher in patients with invasive versus ductal carcinomas in situ. PDGF-CC serum concentrations varied among BC molecular subtypes. Furthermore, we observed a differential expression of most biomarkers between overweight/obese (body mass index (BMI) &ge; 25 kg/m2) and non-obese patients among the BC molecular subtypes. Finally, the classification of subjects according to menopausal status revealed a significant difference in specific biomarker levels between patients and HD. Conclusion: The serum concentrations of angiogenic molecules differ among breast cancer molecular subtypes and are affected by the BMI and menopausal status, which could have possible clinical or prognostic implications

Genotyping KRAS and EGFR Mutations in Greek Patients With Non-small-cell Lung Cancer: Incidence, Significance and Implications for Treatment

Background/Aim: KRAS mutations are reported in 20-25% of non-small cell lung cancer (NSCLC) and their prognostic role is unclear. We studied KRAS and EGFR genotyping in Greek NSCLC patients. Patients and Methods: KRAS and EGFR genotypes were centrally evaluated in 421 NSCLC patients (diagnosed September 1998 -June 2013) and associated with clinicopathological parameters. Outcome comparisons were performed in 288 patients receiving first line treatment. Results: Most patients were male (78.6%), &gt;60 years old (63.9%), current smokers (51.1%), with adenocarcinoma histology (63.9%). EGFR and KRAS mutations were found in 10.7% and 16.6% of all histologies, respectively, and in 14.9% and 21.9% of adenocarcinomas. At 4.5 years median follow-up, KRAS status was an independent negative prognostic factor for overall survival (OS, p=0.016). KRAS mutations conferred 80% increased risk of death in patients receiving first-line treatment (p=0.002). Conclusion: The presence of KRAS mutations is an independent negative prognosticator among Greek NSCLC patients and an independent response predictor to first line treatment