Docosahexaenoic acid (DHA) promotes immunogenic apoptosis in human multiple myeloma cells, induces autophagy and inhibits STAT3 in both tumor and dendritic cells

Docosahexaenoic acid (DHA), a ω-3 polyunsaturated fatty acid found in fish oil, is a multi-target agent and exerts anti-inflammatory and anticancer activities alone or in combination with chemotherapies. Combinatorial anticancer therapies, which induce immunogenic apoptosis, autophagy and STAT3 inhibition have been proposed for long-term therapeutic success. Here, we found that DHA promoted immunogenic apoptosis in multiple myeloma (MM) cells, with no toxicity on PBMCs and DCs. Immunogenic apoptosis was shown by the emission of specific DAMPs (CRT, HSP90, HMGB1) by apoptotic MM cells and the activation of their pro-apoptotic autophagy. Moreover, immunogenic apoptosis was directly shown by the activation of DCs by DHA-induced apoptotic MM cells. Furthermore, we provided the first evidence that DHA activated autophagy in PBMCs and DCs, thus potentially acting as immune stimulator and enhancing processing and presentation of tumor antigens by DCs. Finally, we found that DHA inhibited STAT3 in MM cells. STAT3 pathway, essential for MM survival, contributed to cancer cell apoptosis by DHA. We also found that DHA inhibited STAT3 in blood immune cells and counteracted STAT3 activation by tumor cell-released factors in PBMCs and DCs, suggesting the potential enhancement of the anti-tumor function of multiple immune cells and, in particular, that of DCs

Limited Vitrectomy versus Complete Vitrectomy for Epiretinal Membranes: A Comparative Multicenter Trial

Purpose. To evaluate whether limited vitrectomy is as effective as complete vitrectomy in eyes with epiretinal membrane (ERM) and to compare the surgical times and rates of complications. Methods. In this multicentre European study, data of eyes with ERM that underwent vitrectomy from January 2017 to July 2018 were analyzed retrospectively. In the limited vitrectomy group, a posterior vitreous detachment (PVD) was induced up till the equator as opposed to complete PVD induction till the vitreous base in the comparison group. Incidence of iatrogenic retinal breaks, retinal detachment, surgical time, and visual outcomes were compared between groups. Results. We included 139 eyes in the analysis with a mean age being 72.2 \ub1 6.9 years. In this, sixty-five eyes (47%) underwent limited vitrectomy and 74 eyes (53%) underwent complete vitrectomy. Iatrogenic retinal tears were seen in both groups (5% in limited vitrectomy versus 7% in complete vitrectomy, p=0.49). Retinal detachment occurred in 2 eyes in the limited vitrectomy group (3%) compared to none in the complete vitrectomy group (p=0.22). Best-corrected visual acuity (BCVA) and central macular thickness improved significantly with no intergroup differences (p=0.18). Surgical time was significantly shorter in the limited vitrectomy group with 91% surgeries taking less than 1 hour compared to 71% in the complete vitrectomy group (p<0.001). Conclusion. A limited vitrectomy is a time-efficient and effective surgical procedure for removal of epiretinal membrane with no additional complications

Targeting pancreatic cancer using a combination of gemcitabine with the omega-3 polyunsaturated fatty acid emulsion, Lipidem ™

Scope Pancreatic cancer remains a disease of poor prognosis, with alternate strategies being sought to improve therapeutic efficacy. Omega-3 fatty acids have shown clinical benefit, and mechanisms of action are under investigation. Methods and results Proliferation assays, flow cytometry, invasion assays, ELISA and western blotting were used to investigate efficacy of omega-3 fatty acids alone and in combination with gemcitabine. The docosahexanoic acid (DHA)/eicosapentanoic acid (EPA) combination, Lipidem™, in combination with gemcitabine inhibited growth in pancreatic cancer and pancreatic stellate cell (PSC) lines, with PSCs exhibiting greatest sensitivity to this combination. Invasion of pancreatic cancer cells and PSCs in a 3D spheroid model, was inhibited by combination of gemcitabine with Lipidem™. PSCs were required for cancer cell invasion in an organotypic co-culture model, with invasive capacity reduced by Lipidem™ alone. Platelet-derived growth factor (PDGF) is a key cytokine in pro-proliferative and invasion signalling, and thus a critical regulator of interactions between pancreatic cancer cells and adjacent stroma. Platelet-derived growth factor (PDGF-BB) secretion was completely inhibited by the combination of Lipidem™ with gemcitabine in cancer cells and PSCs. Conclusion Lipidem™ in combination with gemcitabine, has anti-proliferative and anti-invasive efficacy in vitro, with pancreatic stellate cells exhibiting the greatest sensitivity to this combination

Pharmacological and nutritional targeting of voltage-gated sodium channels in the treatment of cancers

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The lymphocyte serine proteinase Granzyme B is expressed in urothelial carcinoma and promotes cancer cell invasion

Dottorato di ricerca in Genetica e biologia cellulareGranzyme B (GrB) è una proteasi di tipo serinico espressa dai linfociti citotossici (linfociti T e cellule NK), che in presenza di perforina (Pf), induce apoptosi nelle cellule bersaglio quali cellule infettate e cellule tumorali. Recentemente, è stato dimostrato che GrB è espressa, spesso in assenza di Pf, anche in cellule non-linfoidi; tuttavia il ruolo di GrB in queste cellule non è stato ancora definito. L’obbiettivo della mia ricerca è stato quello di indagare: - l’espressione di GrB e Pf in linee cellulari e in tessuti di carcinoma della vescica, mediante RT-PCR, Western Blot, ELISA, immunofluorescenza e immunoistochimica. - la funzione di GrB in vitro mediante saggi di attività enzimatica e saggi di invasione cellulare, in presenza o in assenza di silenziamento genico di GrB. I risultati di questo studio hanno rivelato l’espressione di GrB, in assenza di Pf, in cellule e tessuti di carcinoma uroteliale (UC). Differenze significative sono state riscontrate tra l’espressione di GrB e l’aumento dello stadio di invasione del tumore. Di particolare interesse, è stato osservato che l’espressione di GrB in UC è maggiormente concentrata nelle zone di invasione del tumore ed in cellule neoplastiche che vanno incontro a transizione epitelio-mesenchimale (EMT), un evento chiave dell’invasione tumorale. Infatti, cellule tumorali esprimenti GrB co-esprimono marcatori molecolari dell’EMT, quali Snail, N-caderina e perdono l'espressione di E-caderina sulla membrana cellulare. Inoltre, GrB espressa nelle linee cellulari tumorali è enzimaticamente attiva, in quanto cellule tumorali esprimenti GrB contengono un’attività enzimatica specifica di GrB. GrB è capace di tagliare in vitro la vitronectina; questo risultato indica un ruolo di GrB nel rimodellamento della matrice extracellulare (ECM). Infine, abbiamo dimostrato che l'inibizione dell’attività enzimatica di GrB o il silenziamento genico di GrB (ottenuto mediante “RNAinterference”) inibisce significativamente l’invasione su matrigel delle linee cellulari tumorali di vescica. In conclusione, i nostri risultati dimostrano l’espressione di GrB nel carcinoma della vescica e propongono una nuova funzione di GrB, quale la promozione dell’invasione tumorale. L’insieme dei risultati ottenuti da questo studio suggerisce che GrB, mediante la degradazione della ECM, può contribuire all’induzione di un fenotipo invasivo del carcinoma della vescica.Granzyme B (GrB) is a serine proteinase known to be expressed by cytotoxic lymphocytes (T lymphocytes and NK cells) and to induce, in presence of perforin (Pf), apoptosis in target cells, such us infected and tumor cells. Recently, GrB expression has been also shown (often in absence of Pf) in non-lymphoid cells, but its function is not defined. In this study, we investigated: - GrB and Pf expression in bladder cancer cell lines and in bladder cancer tissues by RT-PCR, Western blot, ELISA, immunofluorescence, and immunohistochemistry. - GrB function of in bladder cancer cell lines; the in vitro function of GrB was examined by lossof- function experiments. Our results revealed that GrB is expressed, in absence of Pf, in UC cells. Significant differences were found between GrB expression and both increasing pathological tumor spreading and high grade vs low grade pTa tumors. Notably, GrB in UC tissues was concentrated at the cancer invasion front and was expressed in neoplastic cells undergoing epithelial-mesenchymal transition (EMT), a key event in carcinoma invasion. Indeed, GrB-positive cells also expressed Snail, N-cadherin, or were negative for E-cadherin. Tumor-expressed GrB was enzymatically active and capable of vitronectin cleavage, implying extracellular matrix (ECM) remodeling by GrB. Inhibition of GrB activity or Stealth RNA interference-mediated GrB gene silencing dramatically suppressed bladder cancer cell invasion through matrigel. This data provides the first evidence for a role of GrB in promoting cancer cell invasion. Taken together, our findings suggest that GrB, via ECM degradation, contributes to the establishment of the UC invasive phenotype