RESUMEN
OBJECTIVE: To study the mRNA expression and protein tissue distribution of IL-32 in ileal biopsy specimens from patients with AS. METHODS: Quantitative gene expression analysis, by real-time PCR, of IL-32, IL-1ß, IL-10, TNF-α and IFN-γ was performed on ileal biopsies of 15 AS and 15 Crohn's disease (CD) patients and 10 healthy subjects (HSs). IL-32 tissue distribution was evaluated by immunohistochemistry. The effect of IL-32 on the production of IL-10 by intestinal epithelial cell lines was also evaluated. RESULTS: In the ileal specimens of patients with AS and intestinal chronic inflammation, significant up-regulation of IL-32 at both the mRNA and protein levels was found as compared with non-inflamed AS patients and controls. IL-32 over-expression in AS was accompanied by a significant increase of IL-10 but not of cytokines involved in IL-32 induction. IL-32 stimulates intestinal epithelial cell lines in vitro to produce IL-10. CONCLUSION: Our findings suggest IL-32 as an important cytokine probably involved in the innate immune response occurring in early phases of intestinal inflammation, where it seems to play a prevalent protective role.
Asunto(s)
Enfermedad de Crohn/metabolismo , Íleon/metabolismo , Interleucinas/metabolismo , Espondilitis Anquilosante/metabolismo , Adolescente , Adulto , Estudios de Casos y Controles , Enfermedad de Crohn/inmunología , Células Epiteliales/metabolismo , Femenino , Células HCT116 , Humanos , Ileítis/inmunología , Ileítis/metabolismo , Inmunidad Innata , Interferón gamma/metabolismo , Interleucina-10/biosíntesis , Interleucina-10/metabolismo , Interleucina-1beta/metabolismo , Interleucinas/genética , Macrófagos/metabolismo , Masculino , Persona de Mediana Edad , ARN Mensajero/metabolismo , Espondilitis Anquilosante/inmunología , Factor de Necrosis Tumoral alfa/metabolismo , Adulto JovenRESUMEN
Tumor derived exosomes are vesicles which contain proteins and microRNAs that mediate cell-cell communication and are involved in angiogenesis and tumor progression. Curcumin derived from the plant Curcuma longa, shows anticancer effects. Exosomes released by CML cells treated with Curcumin contain a high amount of miR-21 that is shuttled into the endothelial cells in a biologically active form. The treatment of HUVECs with CML Curcu-exosomes reduced RhoB expression and negatively modulated endothelial cells motility. We showed that the addition of CML control exosomes to HUVECs caused an increase in IL8 and VCAM1 levels, but Curcu-exosomes reversed these effects thus attenuating their angiogenic properties. This antiangiogenic effect was confirmed with in vitro and in vivo vascular network formation assays. SWATH analysis of the proteomic profile of Curcu-exosomes revealed that Curcumin treatment deeply changes their molecular properties, in particular, Curcumin induces a release of exosomes depleted in pro-angiogenic proteins and enriched in proteins endowed with anti-angiogenic activity. Among the proteins differential expressed we focused on MARCKS, since it was the most modulated protein and a target of miR-21. Taken together our data indicated that also Curcumin attenuates the exosome's ability to promote the angiogenic phenotype and to modulate the endothelial barrier organization.
Asunto(s)
Curcumina/farmacología , Exosomas/efectos de los fármacos , Células Endoteliales de la Vena Umbilical Humana/metabolismo , MicroARNs/genética , Antineoplásicos/farmacología , Línea Celular Tumoral , Células Cultivadas , Exosomas/genética , Exosomas/metabolismo , Regulación de la Expresión Génica , Humanos , Células K562 , Leucemia Mielógena Crónica BCR-ABL Positiva/genética , Leucemia Mielógena Crónica BCR-ABL Positiva/metabolismo , Leucemia Mielógena Crónica BCR-ABL Positiva/patología , Sustrato de la Proteína Quinasa C Rico en Alanina Miristoilada/genética , Sustrato de la Proteína Quinasa C Rico en Alanina Miristoilada/metabolismo , Neovascularización Patológica/genética , Neovascularización Fisiológica/genética , Proteoma/genética , Proteoma/metabolismo , Proteómica/métodosRESUMEN
Mutation of the Bcr-Abl oncoprotein is one of most frequent mechanisms by which chronic myelogenous leukemia (CML) cells become resistant to imatinib. Here, we show that treatment of cell lines harbouring wild type or mutant BCR-ABL with carboxyamidotriazole (CAI), a calcium influx and signal transduction inhibitor, inhibits cell growth, the expression of Bcr-Abl and its downstream signalling, and induces apoptosis. Moreover, we show that CAI acts by increasing intracellular ROS. Clinically significant, CAI has also inhibitory effects on T315I Bcr-Abl mutant, a mutation that causes CML cells to become insensitive to imatinib and second generation abl kinase inhibitors.