IGF-IR cooperates with ERα to inhibit breast cancer cell aggressiveness by regulating the expression and localisation of ECM molecules

IGF-IR is highly associated with the behaviour of breast cancer cells. In ERα-positive breast cancer, IGF-IR is present at high levels. In clinical practice, prolonged treatment with anti-estrogen agents results in resistance to the therapy with activation of alternative signaling pathways. Receptor Tyrosine Kinases, and especially IGF-IR, have crucial roles in these processes. Here, we report a nodal role of IGF-IR in the regulation of ERα-positive breast cancer cell aggressiveness and the regulation of expression levels of several extracellular matrix molecules. In particular, activation of IGF-IR, but not EGFR, in MCF-7 breast cancer cells results in the reduction of specific matrix metalloproteinases and their inhibitors. In contrast, IGF-IR inhibition leads to the depletion by endocytosis of syndecan-4. Global important changes in cell adhesion receptors, which include integrins and syndecan-4 triggered by IGF-IR inhibition, regulate adhesion and invasion. Cell function assays that were performed in MCF-7 cells as well as their ERα-suppressed counterparts indicate that ER status is a major determinant of IGF-IR regulatory role on cell adhesion and invasion. The strong inhibitory role of IGF-IR on breast cancer cells aggressiveness for which E2-ERα signaling pathway seems to be essential, highlights IGF-IR as a major molecular target for novel therapeutic strategies

Dynamic interaction between the signaling pathways of ERα, growth factors and extracellular matrix molecules on the aggressive behavior of breast cancer cells

The aggressive behavior and the mesenchymal phenotype are two critical cancer cell properties which are closely associated with tumor development, progression and metastasis. The regulation of these properties in breast cancer is significantly mediated by several signaling molecules, including estrogen receptors and receptors tyrosine kinases. Some ECM molecules, such as proteoglycans and proteolytic enzymes, are also crucial mediators of the above mentioned cellular properties in cancer. Our findings revealed specific molecules and possible mechanisms of action which actively mediate the regulation of aggressiveness and the mesenchymal morphology of breast cancer cells. More specifically, we demonstrated that the silencing of ERα induces EMT and promotes the aggressive behavior of MCF-7 breast cancer cells. In addition, the suppression of ERα activates cell signaling which is responsible for the mediation of aggressiveness, as well as enhances the expression and the activity of a plethora of critical proteolytic molecules. On the other hand, we demonstrated that IGF-IR promotes the mild behavior of breast cancer cells through an ERα-dependent manner, and the same time regulates the expression of specific MMPs/TIMPs. Furthermore, we report that the inhibition of the RTKs AXL and PDGFRβ causes a partial MET and strongly suppresses the high aggressive behavior of MDA-ΜΒ-231 breast cancer cells, thus highlighting the crucial role of these RTKs on the maintenance of mesenchymal phenotype and the aggressiveness in breast cancer. Moreover, the impact of AXL and PDGFRβ on cell signaling, while the crosstalk between the pathways of the RTKs with TGFβ signaling axis was also revealed. Finally, we demonstrated that serglycin mediates the induction of aggressiveness in breast cancer, through the regulation of crucial signaling pathways, including the IL-8/CXCR-2 pathway. Last but not least, it was shown that the overexpression of serglycin enhances the biosynthesis and the activity of the proteolytic network molecules, while it partly induces EMT and confers chemoresistance against commercially available anticancer drugs. Collectively, our data define the decisive regulatory role of all the above mentioned effectors of the aggressive behavior and the mesenchymal phenotype of breast cancer cells, as well as reveal possible mechanisms of action, thus highlighting the complexity for the regulation of these cell properties in breast cancer.Η επιθετική συμπεριφορά και ο μεσεγχυματικός φαινότυπος είναι δυο ιδιότητες των καρκινικών κύτταρων οι οποίες σχετίζονται άμεσα με την ανάπτυξη, την εξέλιξη και τη μετάσταση του όγκου. Στον καρκίνο του μαστού η ρύθμιση αυτών των ιδιοτήτων φαίνεται να διαμεσολαβείται καθοριστικά από διάφορους σηματοδοτικούς άξονες, όπως οι οιστρογονοϋποδοχείς και συγκεκριμένοι υποδοχείς κινασών τυροσίνης. Τα μόρια του ECM, συμπεριλαμβανομένων των πρωτεογλυκανών και των πρωτεολυτικών ενζύμων, είναι επίσης σημαντικοί τελεστές της επαγωγής αυτών των δυο ιδιοτήτων στον καρκίνο. Τα αποτελέσματα της παρούσας μελέτης κατέδειξαν συγκεκριμένα μόρια και πιθανούς μηχανισμούς δράσεις που διέπουν ενεργά τη ρύθμιση της επιθετικότητας και της μεσεγχυματικής μορφολογίας σε καρκινικές σειρές μαστού. Πιο συγκεκριμένα, δείξαμε οτι η καταστολή του ERα στα MCF-7 καρκινικά κύτταρα μαστού επάγει την ΕΜΤ προωθώντας ταυτόχρονα την επιθετική κυτταρική συμπεριφορά. Επιπλέον, η αποσιώπηση του ERα επάγει αισθητά τη σηματοδότηση η οποία διαμεσολαβεί την επιθετική συμπεριφορά, καθώς και ενισχύει σημαντικά την έκφραση και τη δραστικότητα πλήθους κρίσιμων μορίων του πρωτεολυτικού δικτύου. Από την άλλη πλευρά, αναδείχθηκε οτι ο IGF-IR προάγει την ήπια συμπεριφορά των καρκινικών κυττάρων μαστού μέσω μιας διαδικασίας η οποία απαιτεί την παρουσία του ERα, ενώ ρυθμίζει παράλληλα την έκφραση συγκεκριμένων MMPs/TIMPs. Επιπροσθέτως, αναφέρουμε οτι η αναστολή των RTKs AXL και PDGFRβ προκαλεί τη μερική ΜΕΤ και καταστέλλει σημαντικά την υψηλά επιθετική συμπεριφορά των MDA-ΜΒ-231 καρκινικών κυττάρων μαστού, υπογραμμίζοντας τον καθοριστικό ρόλο αυτών των υποδοχέων στη διατήρηση του μεσεγχυματικού φαινοτύπου και της επιθετικότητας στον καρκίνο του μαστού. Επιπλέον, έγινε εμφανής η επίδραση των AXL και PDGFRβ στην κυτταρική σηματοδότηση, καθώς και ανιχνεύθηκε τόσο η μεταξύ τους διεπικοινωνία όσο και η διεπικοινωνία με το TGFβ σηματοδοτικό μονοπάτι. Τέλος, αναδείχθηκε ο καθοριστικός ρόλος της σεργλυκίνης στην επαγωγή της επιθετικότητας στον καρκίνο του μαστού μέσω της ρύθμισης κρίσιμων σηματοδοτικών μονοπατιών, με κυριότερο αυτό της IL-8/CXCR-2. Επιπλέον, έγινε προφανές οτι η υπερέκφραση της σεργλυκίνης ενισχύει την έκφραση και την ενεργότητα των μορίων του πρωτεολυτικού δικτύου, ενώ ταυτόχρονα επάγει μερικώς την ΕΜΤ και προσδίδει στα καρκινικά κύτταρα μαστού ανθεκτικότητα ενάντια σε αντικαρκινικές θεραπείες. Συλλογικά, τα δεδομένα μας διευκρινίζουν τον καθοριστικό ρυθμιστικό ρόλο όλων των παραπάνω τελεστών της επιθετικής συμπεριφοράς και του μεσεγχυματικού φαινοτύπου των καρκινικών κυττάρων μαστού, αναδεικνύοντας παράλληλα πιθανούς μηχανισμούς δράσης και ως εκ τούτου υπογραμμίζοντας την πολυπλοκότητα της ρύθμισης των δυο αυτών κυτταρικών ιδιοτήτων στον καρκίνο του μαστού

Evaluation of Tegaran Formula ZhenHua cytotoxicity against human cancer cell lines.

The aim of this study is to evaluate the potential health effects of Tegaran Formula ZhenHua, a nutritional supplement used mainly by cancer patients. Its active ingredients and cytotoxicity was assessed with analytical methods and viability assays, respectively. The analytical methods consisted of dissolution, disintegration, HPLC, LC/MS, GC/MS and NMR. Cytotoxicity was assessed by MTT, SRB, CVE colorimetric viability assays in 0, 24, 48 and 72h time points. The results indicate that Tegaran Formula ZhenHua supplement did not present any cytotoxic effects due to issues related to the capsules' solubility, distribution and identification of the active ingredient

Estrogen receptor beta modulates breast cancer cells functional properties, signaling and expression of matrix molecules

Estrogen receptors have pivotal roles in breast cancer growth and progression. ER\u3b1 has been clearly shown to play key role in hormone-dependent breast cancer properties, but little is known for the isoform ER\u3b2. To evaluate the role of ER\u3b2, we established stably transfected ER\u3b2-suppressed MDA-MB-231 breast cancer cells by knocking down the human ER\u3b2 gene, using specific shRNA lentiviral particles. As observed by scanning electron microscopy, the ER\u3b2 suppression induces significant phenotypic changes in these cells, as compared to the control cells. Notably, the down-regulation of ER\u3b2 decreases the expression of the mesenchymal markers fibronectin and vimentin, whereas it increases the expression levels of the epithelial marker E-cadherin and cell junctions. These alterations are followed by reduced levels of the functional cell properties that promote the aggressiveness of these cells, such as proliferation, migration, spreading capacity, invasion and adhesion on collagen I. Notably, the down-regulation of ER\u3b2 reduces the migration of breast cancer cells through the tyrosine kinase receptors EGFR/IGF-IR and the JAK/STAT signaling pathways. Moreover, ER\u3b2 has a crucial role on the gene expression of several matrix mediators, including the proteoglycans syndecans-2/-4 and serglycin, several matrix metalloproteinases, plasminogen activation system components and receptor tyrosine kinases. These data clearly show that ER\u3b2 plays a crucial role in the cell behavior and ECM composition of the highly aggressive MDA-MB-231 cells and opens a new area of research to further understand its role and to improve pharmaceutical targeting of the non-hormone dependent breast cancer

The surgical safety checklist and patient outcomes after surgery: a prospective observational cohort study, systematic review and meta-analysis

© 2017 British Journal of Anaesthesia Background: The surgical safety checklist is widely used to improve the quality of perioperative care. However, clinicians continue to debate the clinical effectiveness of this tool. Methods: Prospective analysis of data from the International Surgical Outcomes Study (ISOS), an international observational study of elective in-patient surgery, accompanied by a systematic review and meta-analysis of published literature. The exposure was surgical safety checklist use. The primary outcome was in-hospital mortality and the secondary outcome was postoperative complications. In the ISOS cohort, a multivariable multi-level generalized linear model was used to test associations. To further contextualise these findings, we included the results from the ISOS cohort in a meta-analysis. Results are reported as odds ratios (OR) with 95% confidence intervals. Results: We included 44 814 patients from 497 hospitals in 27 countries in the ISOS analysis. There were 40 245 (89.8%) patients exposed to the checklist, whilst 7508 (16.8%) sustained ≥1 postoperative complications and 207 (0.5%) died before hospital discharge. Checklist exposure was associated with reduced mortality [odds ratio (OR) 0.49 (0.32–0.77); P\u3c0.01], but no difference in complication rates [OR 1.02 (0.88–1.19); P=0.75]. In a systematic review, we screened 3732 records and identified 11 eligible studies of 453 292 patients including the ISOS cohort. Checklist exposure was associated with both reduced postoperative mortality [OR 0.75 (0.62–0.92); P\u3c0.01; I2=87%] and reduced complication rates [OR 0.73 (0.61–0.88); P\u3c0.01; I2=89%). Conclusions: Patients exposed to a surgical safety checklist experience better postoperative outcomes, but this could simply reflect wider quality of care in hospitals where checklist use is routine

Prospective observational cohort study on grading the severity of postoperative complications in global surgery research

Background The Clavien–Dindo classification is perhaps the most widely used approach for reporting postoperative complications in clinical trials. This system classifies complication severity by the treatment provided. However, it is unclear whether the Clavien–Dindo system can be used internationally in studies across differing healthcare systems in high- (HICs) and low- and middle-income countries (LMICs). Methods This was a secondary analysis of the International Surgical Outcomes Study (ISOS), a prospective observational cohort study of elective surgery in adults. Data collection occurred over a 7-day period. Severity of complications was graded using Clavien–Dindo and the simpler ISOS grading (mild, moderate or severe, based on guided investigator judgement). Severity grading was compared using the intraclass correlation coefficient (ICC). Data are presented as frequencies and ICC values (with 95 per cent c.i.). The analysis was stratified by income status of the country, comparing HICs with LMICs. Results A total of 44 814 patients were recruited from 474 hospitals in 27 countries (19 HICs and 8 LMICs). Some 7508 patients (16·8 per cent) experienced at least one postoperative complication, equivalent to 11 664 complications in total. Using the ISOS classification, 5504 of 11 664 complications (47·2 per cent) were graded as mild, 4244 (36·4 per cent) as moderate and 1916 (16·4 per cent) as severe. Using Clavien–Dindo, 6781 of 11 664 complications (58·1 per cent) were graded as I or II, 1740 (14·9 per cent) as III, 2408 (20·6 per cent) as IV and 735 (6·3 per cent) as V. Agreement between classification systems was poor overall (ICC 0·41, 95 per cent c.i. 0·20 to 0·55), and in LMICs (ICC 0·23, 0·05 to 0·38) and HICs (ICC 0·46, 0·25 to 0·59). Conclusion Caution is recommended when using a treatment approach to grade complications in global surgery studies, as this may introduce bias unintentionally

Critical care admission following elective surgery was not associated with survival benefit: prospective analysis of data from 27 countries

This was an investigator initiated study funded by Nestle Health Sciences through an unrestricted research grant, and by a National Institute for Health Research (UK) Professorship held by RP. The study was sponsored by Queen Mary University of London