Specific effects of bortezomib against experimental malignant pleural effusion: a preclinical study

BACKGROUND: We have previously shown that nuclear factor (NF)-κB activation of mouse Lewis lung carcinoma (LLC) specifically promotes the induction of malignant pleural effusions (MPE) by these cells. In the present studies we hypothesized that treatment of immunocompetent mice with bortezomib tailored to inhibit cancer cell NF-κB activation and not proliferation specifically inhibits MPE formation by LLC cells. RESULTS: Treatment of LLC cells with low concentrations of bortezomib (100 ng/ml) inhibited NF-κB activation and NF-κB-dependent transcription, but not cellular proliferation. Bortezomib treatment of immunocompetent C57BL/6 mice bearing LLC-induced subcutaneous tumors and MPEs significantly blocked tumor-specific NF-κB activation. However, bortezomib treatment did not impair subcutaneous LLC tumor growth, but was effective in limiting LLC-induced MPE. This specific effect was evidenced by significant reductions in effusion accumulation and the associated mortality and was observed with both preventive (beginning before MPE formation) and therapeutic (beginning after MPE establishment) bortezomib treatment. The favorable impact of bortezomib on MPE was associated with suppression of cardinal MPE-associated phenomena, such as inflammation, vascular hyperpermeability, and angiogenesis. In this regard, therapeutic bortezomib treatment had identical favorable results on MPE compared with preventive treatment, indicating that the drug specifically counteracts effusion formation. CONCLUSIONS: These studies indicate that proteasome inhibition tailored to block NF-κB activation of lung adenocarcinoma specifically targets the effusion-inducing phenotype of this tumor. Although the drug has limited activity against advanced solid lung cancer, it may prove beneficial for patients with MPE

One-step rapid tracking and isolation of senescent cells in cellular systems, tissues, or animal models via GLF16

Identification and isolation of senescent cells is challenging, rendering their detailed analysis an unmet need. We describe a precise one-step protocol to fluorescently label senescent cells, for flow cytometry and fluorescence microscopy, implementing a fluorophore-conjugated Sudan Black-B analog, GLF16. Also, a micelle-based approach allows identification of senescent cells in vivo and in vitro, enabling live-cell sorting for downstream analyses and live in vivo tracking. Our protocols are applicable to cellular systems, tissues, or animal models where senescence is present. For complete details on the use and execution of this protocol, please refer to Magkouta et al.</p

Effect of mastic oil and its derivatives in angiogenesis

Το μαστιχέλαιο, το αιθέριο έλαιο που προέρχεται από το φυτό Pistacia lentiscus variation chia αποτελεί μη τοξικό φυσικό προϊόν συνδεδεμένο με την παραδοσιακή ιατρική καθώς είναι γνωστό για την αντιμικροβιακή αντιοξειδωτική και αντιφλεγμονώδη του δράση. Τα τελευταία χρόνια έχουν συσσωρευθεί δεδομένα τα οποία αφορούν την αντικαρκινική και αντιαγγειογενετική του δράση. Στόχος της παρούσας διατριβής είναι η διερεύνηση της δράσης του μαστιχέλαιου και των κυριοτέρων συστατικών του στην ανάπτυξη όγκων και την μετάσταση in vivo και να εντοπιστούν οι κυριότεροι μηχανισμοί και τα μόρια στόχοι μέσω των οποίων αυτή πραγματοποιείται. Για το σκοπό αυτό χρησιμοποιήθηκαν δύο ευρέως μελετημένα in vivo μοντέλα, αυτό της υποδόριας εμφύτευσης όγκων σε συγγενή ποντίκια της σειράς C57BL6 για τη μελέτη της αύξησης όγκων και το τροποποιημένο μοντέλο της χοριοαλλαντοϊκής μεμβράνης κοτόπουλου για τη μελέτη της μετάστασης. Χρησιμοποιήθηκαν οι σειρές αδενοκαρκινώματος (LLC) και μελανώματος (Β16) ποντικού. Στο πρώτο μοντέλο παρατηρήθηκε ότι τα ζώα στα οποία χορηγήθηκε μαστιχέλαιο παρουσίασαν παρεμπόδιση της αύξησης των LLC/B16 όγκων κατά στατιστικά σημαντικό τρόπο. Ανοσοϊστοχημική ανάλυση των τομών των όγκων έδειξε ότι η μείωση αυτή σχετίζεται με μειωμένη αγγειογένεση και αυξημένα επίπεδα απόπτωσης στο περιβάλλον του όγκου. Επιπλέον τα επίπεδα των προφλεγμονωδών πρωτεϊνών Χημειοτακτική Πρωτεΐνη Μονοκυττάρων 1 (Monocyte Chemoattractant Protein 1 MCP 1) και Φλεγμονώδους Πρωτεΐνης Μακροφάγων 2 (Macrophage Inflammatory Protein 2 ΜΙΡ 2) ήταν χαμηλότερα σε σχέση με εκείνα των ζωών στα οποία χορηγήθηκε ο φορέας. Η δράση του μαστιχέλαιου σχετίζεται περαιτέρω με τη μείωση των συνολικών επίπεδων έκφρασης των μικρών GTPασων RhoA και Ras που αποτελούν μόρια κλειδιά μεταγωγής σημάτων και τη μειωμένη ενεργοποίηση του γενικού μεταγραφικού παράγοντα NF-κB in vivo αλλά και in vitro. Στο τροποποιημένο μοντέλο της χοριοαλλαντοϊκής μεμβράνης κοτόπουλου βρέθηκε ότι το μαστιχέλαιο παρεμποδίζει τη μετάσταση των καρκινικών κυττάρων στο κάτω μέρος της μεμβράνης μειώνοντας ταυτόχρονα την αγγειογένεση στο πάνω μέρος της. Η αντιμεταστατική αυτή δράση του μαστιχέλαιου σχετίζεται περαιτέρω με την ικανότητά του να παρεμποδίζει τη διήθηση τη μετανάστευση και την προσκόλληση των καρκινικών κυττάρων στην εξωκυττάρια ύλη και την έκκριση της Μεταλλοπρωτεϊνάσης υποστρώματος 2 (Matrix Metalloproteinase 2 ΜΜΡ 2). Η δράση του αυτή σχετίζεται με την ικανότητά του να παρεμποδίζει την αναδιοργάνωση του κυτταροσκελετού μέσω της μείωσης του πολυμερισμού της ακτίνης στα καρκινικά κύτταρα. Επιπλέον, μειώνει την ικανότητα προσκόλλησης των καρκινικών κυττάρων στο ενδοθήλιο μέσω της αναστολής της έκφρασης των μορίων προσκόλλησης ICAM 1 και VCAM 1 από τα ενδοθηλιακά κύτταρα και παρεμποδίζει τη μεταναστευτική απόκριση των ενδοθηλιακών κυττάρων σε χημειοτακτικους παράγοντες που εκκρίνονται από τα καρκινικά κύτταρα. Για τους παραπάνω λόγους φαίνεται ότι το μαστιχέλαιο έχει αντικαρκινική και αντιμεταστατική δράση in vivo και αποτελεί ένα χρήσιμο εργαλείο στο πεδίο της χημειοπροστασίας

Mammalian target of rapamycin (mTOR) inhibition does not prevent lung adenocarcinoma-induced malignant pleural effusion

The impact of temsirolimus was investigated in a murine model of malignant pleural effusion (MPE) created with intrapleural injection of Lewis Lung Cancer (LLC) cells. Temsirolimus (1 or 20 mg/kg) did not affect the pleural fluid volume or the number of pleural tumour foci. In addition, temsirolimus did not affect vascular endothelial growth factor expression by LLC cells in vitro. In conclusion, temsirolimus did not curtail experimental lung-adenocarcinoma- induced MPE. © 2013 The Authors. Respirology © 2013 Asian Pacific Society of Respirology

Mastic oil inhibits the metastatic phenotype of mouse lung adenocarcinoma cells

Mastic oil from Pistacia lentiscus variation chia, a natural combination of bioactive terpenes, has been shown to exert anti-tumor growth effects against a broad spectrum of cancers including mouse Lewis lung adenocarcinomas (LLC). However, no studies have addressed its anti-metastatic actions. In this study, we showed that treatment of LLC cells with mastic oil within a range of non-toxic concentrations (0.01-0.04% v/v): (a) abrogated their Matrigel invasion and migration capabilities in transwell assays; (b) reduced the levels of secreted MMP-2; (c) restricted phorbol ester-induced actin remodeling and (d) limited the length of neo-vessel networks in tumor microenvironment in the model of chick embryo chorioallantoic membrane. Moreover, exposure of LLC and endothelial cells to mastic oil impaired their adhesive interactions in a co-culture assay and reduced the expression of key adhesion molecules by endothelial cells upon their stimulation with tumor necrosis factor-alpha. Overall, this study provides novel evidence supporting a multipotent role for mastic oil in prevention of crucial processes related to cancer metastasis. © 2011 by the authors; licensee MDPI, Basel, Switzerland

Mastic Oil Inhibits the Metastatic Phenotype of Mouse Lung Adenocarcinoma Cells

Mastic oil from Pistacia lentiscus variation chia, a natural combination of bioactive terpenes, has been shown to exert anti-tumor growth effects against a broad spectrum of cancers including mouse Lewis lung adenocarcinomas (LLC). However, no studies have addressed its anti-metastatic actions. In this study, we showed that treatment of LLC cells with mastic oil within a range of non-toxic concentrations (0.01–0.04% v/v): (a) abrogated their Matrigel invasion and migration capabilities in transwell assays; (b) reduced the levels of secreted MMP-2; (c) restricted phorbol ester-induced actin remodeling and (d) limited the length of neo-vessel networks in tumor microenvironment in the model of chick embryo chorioallantoic membrane. Moreover, exposure of LLC and endothelial cells to mastic oil impaired their adhesive interactions in a co-culture assay and reduced the expression of key adhesion molecules by endothelial cells upon their stimulation with tumor necrosis factor-alpha. Overall, this study provides novel evidence supporting a multipotent role for mastic oil in prevention of crucial processes related to cancer metastasis

Synthesis of amphipathic derivatives of perillyl glucoside by lipase-catalysed esterification in non conventional media

Perillyl alcohol (POH), is a plant secondary metabolite. It is found in small concentrations in the essential oils of Chios mastic gum, lavendin, peppermint, spearmint etc. Perillyl alcohol (POH) is a monoterpene with anticarcinogenic and antitumor activity in murine tumor models and has been reported to possess chemotherapeutic activity against malignancies. Perillyl glucoside is also a natural compound contained in Perilla frutescens but no data about its biological activity have ever been reported. The studies of lipophilicity and hydrolysis in vitro of prodrugs verified that drugñ saccharide derivatives had amphipathic properties and both lipophilic and amphipathic drug derivatives had obvious controlled release characteristics. Some amphipathic macromolecule drugs are designed for improving pharmaceuticals delivery properties and generally used to enhance the permeability of cell membranes. In the present study is demonstrated the enzymatic synthesis of a perillyl glucoside fatty ester. In the first step, almond β- glucosidase catalyzed the glucosidation of perillyl alcohol by reverse hydrolysis with β-glucose in a biphasic system. In the second step, immobilised lipase from Candida antarctica (Novozym 435) was used in organic media to catalyse the acylation of perillyl glucoside with a medium chain fatty acid. The chemical structure of the products was determined by Mass Spectrometry analysis (MS). The antiproliferative activity of perillyl glucoside and perillyl glucoside fatty ester on LLC cells was investigated. Acknowledgements: This work has been funded by the project PENED 2003. The project is cofinanced 75% of public expenditure through EC - European Social Fund, 25% of public expenditure through Ministry of Development - General Secretariat of Research and Technology and through private sector, under measure 8.3 of OPERATIONAL PROGRAMME "COMPETITIVENESS" in the 3rd Community Support Programme

Temsirolimus targets multiple hallmarks of cancer to impede mesothelioma growth in vivo

Background and objective The mechanistic target of rapamycin (mTOR) promotes cancer cell proliferation and survival, transduces pro-angiogenic signals and regulates immune cell differentiation and function. We hypothesized that temsirolimus, an mTOR inhibitor, would curtail experimental mesothelioma progression in vivo by limiting tumour cell growth, abrogating tumour angiogenesis and modulating immune/inflammatory tumour milieu. Methods We produced flank and pleural syngeneic murine mesotheliomas by delivering AE17 and AB1 murine mesothelioma cells into the right flank or the pleural space of C57BL/6 and BALB/c mice, respectively. Animals were given five times/week intraperitoneal injections of 20-mg/kg temsirolimus or vehicle and were sacrificed on day 26 (flank) or on day 15 (pleural) post-tumour cell propagation. Results Temsirolimus limited mesothelioma growth in vivo by stimulating tumour cell apoptosis, inhibiting tumour angiogenesis, enhancing tumour lymphocyte abundance and blocking pro-tumour myeloid cell recruitment. Pleural fluid accumulation was significantly mitigated in AE17 but not in AB1 mesotheliomas. In vitro, temsirolimus hindered mesothelioma cell growth, NF-kappaB activation and macrophage migration. Conclusions In conclusion, temsirolimus apart from inducing tumour cell apoptosis, targets tumour angiogenesis and influences inflammatory tumour microenvironment to halt experimental mesothelioma growth in vivo. We used syngeneic in vivo mesothelioma models to show that temsirolimus, an mTOR inhibitor, curtailed mesothelioma growth by inducing tumour cell apoptosis, by targeting tumour angiogenesis and by modulating host immune response. This is the first report on the effects of temsirolimus impact on mesothelioma-related immune response and mesothelioma vascularization. © 2015 Asian Pacific Society of Respirology