Remarkable deregulation of microRNAs has been demonstrated in epithelial ovarian cancer (EOC). In particular, some of the let-7 miRNA family members have been proposed as tumor suppressors. Here, we explored the functional roles of let-7g in EOC. The ectopic overexpression of let-7g in OVCAR3 and HEY-A8 EOC cells induced i) a down-regulation of c-Myc and cyclin-D2 thus promoting cell cycle arrest, ii) a reduction of Vimentin, Snail and Slug thus counteracting the progression of epithelial to mesenchymal transition, iii) a chemosensitization to cis -platinum treatment. Next, analysis of human EOC tissues revealed that let-7g expression was significantly reduced in tumor tissue specimens of patients with EOC compared to their non-tumor counterparts ( p = 0.0002). Notably, low let-7g tissue levels were significantly associated with acquired chemoresistance of patients with late-stage of EOC (n = 17, p = 0.03194). This finding was further validated in the serum samples collected from the same cohort of patients (n = 17, p = 0.003). To conclude, we demonstrate that let-7g acts as tumor suppressor and might be used to disable EOC tumor progression and chemoresistance to cis -platinum-based chemotherapy. Furthermore, we propose that decreased expression of let-7g could serve as a tissue and serum biomarker able to predict the chemo-resistant features of EOC patients.

Cardiac myxoma is the most common benign cardiac tumour, localized generally in the left atrium. The majority of cardiac myxomas occur sporadically, while a relatively small proportion of cases develop as a part of Carney complex syndrome. Currently, the histogenesis of myxoma is poorly understood; however, the mesenchymal and endothelial properties of myxoma cells suggest that a clearer understanding of tumour origins can be achieved through a detailed investigation of heart development and endocardial histogenesis. Growing evidence appears to indicate the reactivated expression in cardiac myxoma of genes encoding heart precursor markers, although the exact mechanisms have not yet been described. In this paper we review the most recent scientific literature concerning cardiac embryology and relate this to recent advances in our understanding of the histogenesis of cardiac myxoma. Moreover, given that much of the literature regarding myxoma is of single case reports, we review progress in our knowledge of the pathology and pathogenesis of this condition.

BackgroundCirculating tumor cells (CTCs) represent one of the most interesting target in improving diagnosis, prognosis and treatment. Herein we evaluate the possibility of using an emo-cytometric approach on the evaluation of the heterogeneous population of CTCs to improve personalized metastatic risk assessment. We benchmarked ex vivo behavior of distinct subsets of circulating colon tumor cells with correspondent clinical behavior of patients from which we isolated CTCs.MethodsIsolation and CTC expansion were performed by a gradient protocol. In vitro characterization was determined by flow cytometry, immunofluorescence, western blotting and proteomic profiling. Cell sorter was performed with immunomagnetic beads. Confocal microscopy was used to evaluate tissue sections. Kaplan Mayer curves was cared for through Medcalc program.ResultsWe collected heterogeneous CTCs, derived from the whole blood of seven patients affected by colon cancer, expressing CD133posCD45neg (5 ± 1) and (2 ± 1) and CK20posCD45neg of (29 ± 3) (11 ± 1) cells/ml in Dukes D and A stage respectively. Proliferation rate of 57 ± 16 %, expression for CXCR4pos of 18 ± 7 % and detectable levels of IL-6, IL-8 and SDF-1 cytokines in conditioned culture medium characterized short-time expanded–CTCs (eCTCs). ECTCs organized in tumor sphere were CD45negCD133pos while in adhesion were CXCR4posCK20pos. These two subsets were separately injected in mice. The first group of xenografts developed superficial lesions within 2 weeks. In the second group, in absence of growing tumour, the survival of injected eCTCs was monitored through SDF-1 serum levels detection. The detection of human cancer cells expressing CK20, in mice tissues sections, suggested a different biological behaviour of injected eCTC-subsets: tumorigenic for the first and disseminating for the second. The benchmarking of the experimental data with the clinical course highlights that patients with prevalence of circulating cancer stem cells (CD45negCD133pos) have a lower overall survival. Conversely, patients with prevalence of circulating differentiated cells (CXCR4posCK20pos) have a low disease-free survival.ConclusionOn the basis of the heterogeneous composition and despite the low number of CTCs, it was possible to distinguish two subgroups of CTCs, suggesting a different clinical outcome. CTC-subsets detailing is useful to better define the metastatic–risk personalized score thus improving disease management and reducing patient care cost.Electronic supplementary materialThe online version of this article (doi:10.1186/s12967-016-0876-y) contains supplementary material, which is available to authorized users.

Dendritic cells (DCs) have a key role in regulating tumor immunity, tumor cell growth and drug resistance. We hypothesized that multiple myeloma (MM) cells might recruit and reprogram DCs to a tumor-permissive phenotype by changes within their microRNA (miRNA) network. By analyzing six different miRNA-profiling data sets, miR-29b was identified as the only miRNA upregulated in normal mature DCs and significantly downregulated in tumor-associated DCs. This finding was validated in primary DCs co-cultured in vitro with MM cell lines and in primary bone marrow DCs from MM patients. In DCs co-cultured with MM cells, enforced expression of miR-29b counteracted pro-inflammatory pathways, including signal transducer and activator of transcription 3 and nuclear factor-κB, and cytokine/chemokine signaling networks, which correlated with patients’ adverse prognosis and development of bone disease. Moreover, miR-29b downregulated interleukin-23 in vitro and in the SCID-synth-hu in vivo model, and antagonized a Th17 inflammatory response. All together, these effects translated into strong anti-proliferative activity and reduction of genomic instability of MM cells. Our study demonstrates that MM reprograms the DCs functional phenotype by downregulating miR-29b whose reconstitution impairs DCs ability to sustain MM cell growth and survival. These results underscore miR-29b as an innovative and attractive candidate for miRNA-based immune therapy of MM.

Bisphosphonates (BPs) are widely used to treat several metabolic and oncological diseases affecting the skeletal system. Despite BPs’ well-known therapeutic potential, they also displayed important side effects, among which is BPs-related osteonecrosis of the jaw, by targeting osteoclast activities, osteoblast, and osteocyte behavior. The aim of this study is to evaluate the biological effects of zoledronic acid (ZOL) in an in vitro model of periodontal ligament stem cells (PDLSCs) by using an experimental setting that resembles the in vivo conditions. PDLSCs were treated with different concentrations of ZOL ranging from 0.1 to 5 μM. The effects of ZOL exposure were evaluated on cell viability via 3-[4,5-Dimethylthiaoly]-2,5-diphenyltetrazolium bromide (MTT), cell cycle analysis, apoptosis detection, and immunofluorescence. Quantitative real-time polymerase chain reaction (PCR), colorimetric detection of alkaline phosphatase activity, and Alizarin Red S staining were performed to investigate the osteogenic potential of PDLSCs exposed to ZOL. MTT analysis showed that the viability of PDLSCs exposed to ZOL concentration ≥1.5 μM for 3 and 6 days was significantly lower ( P < 0.001) than that of untreated cells. The percentage of apoptotic cells was significantly higher in PDLSCs exposed for 4 days to ZOL at 2 μM ( P < 0.01) and 5 μM ( P < 0.001) when compared to the control. Moreover, ZOL treatment (3 days) accounted for alterations in cell cycle distribution, with an increase in the proportion of cells in G0/G1 phase and a reduction in the proportion of cells in S phase. Chronic exposure (longer than 7 days) of PDLSCs to ZOL accounted for the downregulation of ALP, RUNX2, and COL1 genes at all tested concentrations, which fit well with the reduced alkaline phosphatase activity reported after 7 and 14 days of treatment. Reduced Col1 deposition in the extracellular matrix was reported after 14 days of treatment. Increased calcium deposits were observed in treated cells when compared to the control cultures. In conclusion, chronic exposure to 1 μM ZOL induced significant reduction of osteogenic differentiation, while ZOL concentrations ≥1.5 μM are required to impair PDLSCs viability and induce apoptosis.

Reactive oxygen species (ROS) mediates cisplatin-induced cytotoxicity in tumor cells. However, when cisplatin-induced ROS do not reach cytotoxic levels, cancer cells may develop chemoresistance. This phenomenon can be attributed to the inherited high expression of antioxidant protein network. H-Ferritin is an important member of the antioxidant system due to its ability to store iron in a nontoxic form. Altered expression of H-Ferritin has been described in ovarian cancers; however, its functional role in cisplatin-based chemoresistance of this cancer type has never been explored. Here, we investigated whether the modulation of H-Ferritin might affect cisplatin-induced cytotoxicity in ovarian cancer cells. First, we characterized OVCAR3 and OVCAR8 cells for their relative ROS and H-Ferritin baseline amounts. OVCAR3 exhibited lower ROS levels compared to OVCAR8 and greater expression of H-Ferritin. In addition, OVCAR3 showed pronounced growth potential and survival accompanied by the strong activation of pERK/pAKT and overexpression of c-Myc and cyclin E1. When exposed to different concentrations of cisplatin, OVCAR3 were less sensitive than OVCAR8. At the lowest concentration of cisplatin (6 μM), OVCAR8 underwent a consistent apoptosis along with a downregulation of H-Ferritin and a consistent increase of ROS levels; on the other hand, OVCAR3 cells were totally unresponsive, H-Ferritin was almost unaffected, and ROS amounts met a slight increase. Thus, we assessed whether the modulation of H-Ferritin levels was able to affect the cisplatin-mediated cytotoxicity in both the cell lines. H-Ferritin knockdown strengthened cisplatin-mediated ROS increase and significantly restored sensitivity to 6 μM cisplatin in resistant OVCAR3 cells. Conversely, forced overexpression of H-Ferritin significantly suppressed the cisplatin-mediated elevation of intracellular ROS subsequently leading to a reduced responsiveness in OVCAR8 cells. Overall, our findings suggest that H-Ferritin might be a key protein in cisplatin-based chemoresistance and that its inhibition may represent a potential approach for enhancing cisplatin sensitivity of resistant ovarian cancer cells.

The periodontal ligament displays a reservoir of mesenchymal stem cells which can account for periodontal regeneration. Despite the numerous studies directed at the definition of optimal culture conditions for long-term expansion of periodontal ligament stem cells (PDLSCs), no consensus has been reached as to what is the ideal protocol. The aim of the present study was to determine the optimal medium formulation for long-term expansion and stemness maintenance of PDLSCs, in order to obtain a sufficient number of cells for therapeutic approaches. For this purpose, the effects of three different culture medium formulations were evaluated on PDLSCs obtained from three periodontal ligament samples of the same patient: minimum essential medium Eagle, alpha modification (α-MEM), Dulbecco’s modified Eagle’s medium (DMEM), both supplemented with 10% fetal bovine serum (FBS), and a new medium formulation, Ham’s F12 medium, supplemented with 10% FBS, heparin 0.5 U/ml, epidermal growth factor (EGF) 50 ng/ml, fibroblast growth factor (FGF) 25 ng/ml, and bovine serum albumin (BSA) 1% (enriched Ham’s F12 medium; EHFM). PDLSCs grown in EHFM displayed a higher PE-CD73 mean fluorescence intensity compared with cells maintained in α-MEM and DMEM, even at later passages. Cells maintained in EHFM displayed an increased population doubling and a reduced population doubling time compared with cells grown in DMEM or α-MEM. α-MEM, DMEM and EHFM with added dexamethasone, 2-phospho-L-ascorbic acid, and β-glycerophosphate were all able to promote alkaline phosphatase activity; however, no calcium deposition was detected in PDLSCs cultured in EHFM-differentiation medium. When EHFM-, α-MEM- and DMEM-expanded PDLSCs were transferred to a commercial culture medium for the osteogenesis, mineralization became much more evident in confluent monolayers of EHFM-expanded PDLSCs compared with DMEM and α-MEM. The results suggest EHFM is the optimal medium formulation for growth and stemness maintenance of primary PDLSCs. Moreover, EHFM confers higher osteogenic potential to PDLSCs compared with cells maintained in the other culture media. Overall, the results of the present work confirmed the advantages of using EHFM for long-term expansion of mesenchymal cells in vitro and the preservation of high osteogenic potential.

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