Search

BackgroundETV4 is one of the ETS proteins overexpressed in prostate cancer (PC) as a result of recurrent chromosomal translocations. In human prostate cell lines, ETV4 promotes migration, invasion, and proliferation; however, its role in PC has been unclear. In this study, we have explored the effects of ETV4 expression in the prostate in a novel transgenic mouse model.MethodsWe have created a mouse model with prostate-specific expression of ETV4 (ETV4 mice). By histochemical and molecular analysis, we have investigated in these engineered mice the expression of p21, p27, and p53. The implications of our in vivo findings have been further investigated in human cells lines by chromatin-immunoprecipitation (ChIP) and luciferase assays.ResultsETV4 mice, from two independent transgenic lines, have increased cell proliferation in their prostate and two-thirds of them, by the age of 10 months, developed mouse prostatic intraepithelial neoplasia (mPIN). In these mice,cdkn1aand its p21 protein product were reduced compared to controls; p27 protein was also reduced. By ChIP assay in human prostate cell lines, we show that ETV4 binds to a specific site (-704/-696 bp upstream of the transcription start) in theCDKN1Apromoter that was proven, by luciferase assay, to be functionally competent. ETV4 further controlsCDKN1Aexpression by downregulating p53 protein: this reduction of p53 was confirmed in vivo in ETV4 mice.ConclusionsETV4 overexpression results in the development of mPIN but not in progression to cancer. ETV4 increases prostate cell proliferation through multiple mechanisms, including downregulation ofCDKN1Aand its p21 protein product: this in turn is mediated through direct binding of ETV4 to theCDKN1Apromoter and through the ETV4-mediated decrease of p53. This multi-faceted role of ETV4 in prostate cancer makes it a potential target for novel therapeutic approaches that could be explored in this ETV4 transgenic model.

Simple Summary Metastatic melanoma is one of the most aggressive and lethal malignancies with a poor prognosis. Several data underline the crucial role of amoeboid motility in the dissemination of metastatic melanoma cells. Thus, targeting this phenomenon could represent a promising strategy to prevent metastasis formation and improve metastatic melanoma patient’s survival. With this aim, we investigated the effect of Claisened Hexafluoro, a chemical analogue of Honokiol, on human metastatic melanoma cells. We demonstrated that Claisened Hexafluoro, by deregulating the mitochondrial activity, inhibits amoeboid motility and impairs many steps of the dissemination process, finally decreasing the in vivo metastatic spreading. Collectively, these data suggest possible future applications of Claisened Hexafluoro for the treatment of metastatic melanoma. Abstract Metastatic melanoma is characterized by poor prognosis and a low free-survival rate. Thanks to their high plasticity, melanoma cells are able to migrate exploiting different cell motility strategies, such as the rounded/amoeboid-type motility and the elongated/mesenchymal-type motility. In particular, the amoeboid motility strongly contributes to the dissemination of highly invasive melanoma cells and no treatment targeting this process is currently available for clinical application. Here, we tested Claisened Hexafluoro as a novel inhibitor of the amoeboid motility. Reported data demonstrate that Claisened Hexafluoro specifically inhibits melanoma cells moving through amoeboid motility by deregulating mitochondrial activity and activating the AMPK signaling. Moreover, Claisened Hexafluoro is able to interfere with the adhesion abilities and the stemness features of melanoma cells, thus decreasing the in vivo metastatic process. This evidence may contribute to pave the way for future possible therapeutic applications of Claisened Hexafluoro to counteract metastatic melanoma dissemination.

In this manuscript, we show the involvement of the uPA/uPAR system in the regulation of aerobic glycolysis of melanoma cells. uPAR over-expression in human melanoma cells controls an invasive and glycolytic phenotype in normoxic conditions. uPAR down-regulation by siRNA or its uncoupling from integrins, and hence from integrin-linked tyrosine kinase receptors (IL-TKRs), by an antagonist peptide induced a striking inhibition of the PI3K/AKT/mTOR/HIF1α pathway, resulting into impairment of glucose uptake, decrease of several glycolytic enzymes and of PKM2, a checkpoint that controls metabolism of cancer cells. Further, binding of uPA to uPAR regulates expression of molecules that govern cell invasion, including extracellular matrix metallo-proteinases inducer (EMPPRIN) and enolase, a glycolytyc enzyme that also serves as a plasminogen receptor, thus providing a common denominator between tumor metabolism and phenotypic invasive features. Such effects depend on the α5β1-integrin-mediated uPAR connection with EGFR in melanoma cells with engagement of the PI3K-mTOR-HIFα pathway. HIF-1α trans-activates genes whose products mediate tumor invasion and glycolysis, thus providing the common denominator between melanoma metabolism and its invasive features. These findings unveil a unrecognized interaction between the invasion-related uPAR and IL-TKRs in the control of glycolysis and disclose a new pharmacological target (i.e., uPAR/IL-TKRs axis) for the therapy of melanoma.

Introduction The use of movies in medical (particularly psychiatric) education has been often limited to portraits of mental illness and psychiatrists. The Psychiatric Institute of the Università del Piemonte Orientale has a longstanding tradition of working with/on movies according to a method developed by Eugenio Torre, using dynamic images as educational incitements. Our aim is to describe the preliminary results on the impact of this intervention in medical students. Methods The cinemeducation project lasted 6 months, and included 12 meetings. Forty randomly selected participants were assessed with: Attitudes Towards Psychiatry Scale (ATP-30), Social Distance Scale (SDS), Interpersonal Reactivity Index (IRI), and Toronto Alexithymia Scale (TAS), both at baseline and after 6 months, when the workshop was concluded. Results A significant increase was found in the ATP-30 score, and a reduction of the SDS and IRI-Personal Distress scale scores. Conclusions Informal feedback from participants was strongly positive. Preliminary results from the assessment of participants are encouraging. Students’ attitudes towards psychiatry and ability to tolerate anxiety when experiencing others’ distress improved, while stigma decreased. The evocative power of movie dynamic images, developed in the group and integrated with the help of the group leader, can enrich students’ knowledge, both from a cognitive and emotional standpoint.

Introduction Psychiatric management of inpatients with suicidal behavior involves psychiatrists and psychiatric nurses. Specific precautions should be adopted: observation on a one to one basis with the cooperation of qualified nurses and accommodating the patient in a room close to the infirmary is usually recommended. An ongoing assessment of suicidal risk is warranted to check the response to treatment over time. Aim To compare the severity of depressive symptoms in depressed inpatients admitted after an attempted suicide and in depressed inpatients admitted for any other reason; to assess the severity of attempted suicide and the management of suicidal risk in the clinical setting. Material and method We divided the sample in two subgroups: patients with a diagnosis of depression admitted for a current suicide attempt and depressed patients without a current suicide attempt. Socio-demographic and clinical features were gathered; assessment included the Montgomery-Asberg Depression Rating Scale (MADRS) and the Nurses’ Global Assessment of Suicide Risk (NGASR). Results 46 patients were recruited in a 1-year period and were eligible for our study: 20 had been admitted to the hospital after attempting suicide, while the other 26 had not attempted suicide and were admitted for other reasons related to their depression. The multivariate analysis yielded a statistically significant correlation between the use of antidepressants and current suicide attempt. The occurrence of suicidal attempt was not significantly related to the severity of depressive symptoms. In the current suicide attempters subgroup, NGASR suicide risk levels at discharge were significantly lower than those measured at admission. Current suicide attempters had a higher number of suicide attempts in their clinical history, compared to the other group. Conclusion We found no significant correlation among psychiatric diagnosis, severity of depressive symptoms and current suicide attempt. Antidepressant therapy was found to play the role of protective factor for suicide attempts. History of suicide attempts was one of the major predictor for reattempted suicide. A more thorough understanding of the complex phenomenon of suicide and of the reasons underlying suicidal behaviors is warranted.

This work describes the effectiveness of HDAC-inhibitor (S)-2 towards colorectal cancer (CRC) HCT116 cells in vitro by inducing cell cycle arrest and apoptosis, and in vivo by contrasting tumour growth in mice xenografts. Among the multifaceted drug-induced events described herein, an interesting link has emerged between the oncoprotein histone deacetylase HDAC1 and the oncogenic Cancerous Inhibitor of Protein Phosphatase 2A (CIP2A) which is overexpressed in several cancers including CRCs. HDAC1 inhibition by (S)-2 or specific siRNAs downregulates CIP2A transcription in three different CRC cell lines, thus restoring the oncosuppressor phosphatase PP2A activity that is reduced in most cancers. Once re-activated, PP2A dephosphorylates pGSK-3β(ser9) which phosphorylates β-catenin that remains within the cytosol where it undergoes degradation. The decreased amount/activity of β-catenin transcription factor prompts cell growth arrest by diminishing c-Myc and cyclin D1 expression and abrogating the prosurvival Wnt/β-catenin signaling pathway. These results are the first evidence that the inhibition of HDAC1 by (S)-2 downregulates CIP2A transcription and unleashes PP2A activity, thus inducing growth arrest and apoptosis in CRC cells.

Introduction Half of cases of Prostate cancer (PC) carry a chromosomal translocation involving one of the genes of the ETS family (ERG, ETV1, ETV4, ETV5, FLI1). Each translocation results in the deregulated expression of an ETS protein in the prostate that, likely, plays a direct role in PC pathogenesis. We have showed that in prostate cell lines ETV4 overexpression increases migration and invasion, and also proliferation, cell cycle progression, anchorage-independent growth, tumor growth in xenograft model. Here we have investigated the effects of ETV4 expression in mouse prostate. Material and Methods The truncated form of ETV4, present in patients, has been cloned under the control a modified rat probasin gene promoter that drives prostate specific expression. By using this construct we have obtained two different transgenic mouse lines with ETV4 specific prostate expression (ETV4-FVB mice). The tissues of these mice have been analyzed by H&E staining and immunohistochemistry. Result and discussion 6 months-old (mo) ETV4-FVB mice did not present prostate lesions, whereas low grade prostatic intraepithelial neoplasia (LGPIN) lesions have been found in about half of 10 mo mice (10 of 18 line C, 4 of 11 line E) and in none of control mice. Since Pten loss is common in PC and it is often associated with ETS translocations, we have investigated the interaction of ETV4 expression with Pten loss by crossing our FVB-ETV4 mice with PTEN+/- mice. Analyzing the progeny of this cross at 7 months of age we found LGPIN in 33% of FVB-ETV4 mice and in 50% of Pten+/- mice, whereas we found multifocal high grade PIN (HGPIN) lesions in all Pten+/-,ETV4 mice (n=9). In addition, at least one LGPIN was found in all 2 months-old Pten+/-,ETV4. Thus, Pten+/-,ETV4 developed early neoplastic lesions whose number and severity increased with age: in fact, HGPIN progressed to locally invasive adenocarcinoma in 12.5% of 7 mo mice and in 50% 10 mo mice. Conclusion These results suggest that ETV4 over-expression is sufficient to initiate neoplastic transformation and cooperates with Pten loss in promoting prostate cancer progression. Thus, also in vivo, ETV4 induces most and perhaps all of the features that make a tumor. Now we are investigating the molecular mechanisms through which ETV4 cooperates with Pten loss to promote cancer progression. These mouse models, in addition to provide insight into PC biology, will be useful to test the effectiveness of experimental therapeutic approaches.

We have previously shown that peculiar metabolic features of cell adaptation and survival in hypoxia imply growth restriction points that are typical of embryonic stem cells and disappear with differentiation. Here we provide evidence that such restrictions can be exploited as specific antiblastic targets by physiological factors such as pyruvate, tetrahydrofolate, and glutamine. These metabolites act as powerful cytotoxic agents on cancer stem cells (CSCs) when supplied at doses that perturb the biochemical network, sustaining the resumption of aerobic growth after the hypoxic dormant state. Experiments were performed in vivo and in vitro using CSCs obtained from various anaplastic tumors: human melanoma, leukemia, and rat hepatoma cells. Pretreatment of melanoma CSCs with pyruvate significantly reduces their self-renewal in vitro and tumorigenicity in vivo. The metabolic network underlying the cytotoxic effect of the physiological factors was thoroughly defined, principally using AH130 hepatoma, a tumor spontaneously reprogrammed to the embryonic stem stage. This network, based on a tight integration of aerobic glycolysis, cellular redox state, and folate metabolism, is centered on the cellular NADP/NADPH ratio that controls the redox pathway of folate utilization in purine synthesis. On the whole, this study indicates that pyruvate, FH 4, and glutamine display anticancer activity, because CSCs are committed to survive and maintain their stemness in hypoxia. When CSC need to differentiate and proliferate, they shift from anaerobic to aerobic status, and the few mitochondria available makes them susceptible to the injury of the above physiological factors. This vulnerability might be exploited for novel therapeutic treatments.

BackgroundTGFß overproduction in cancer cells is one of the main characteristics of late tumor progression being implicated in metastasis, tumor growth, angiogenesis and immune response. We investigated the therapeutic efficacy of anti-TGFß peptides in the control of angiogenesis elicited by conditional over-expression of TGFß.MethodsWe have inserted in human MCF7 mammary-cancer cells a mutated TGFß gene in a tetracycline-repressible vector to obtain conditional expression of mature TGFß upon transient transfection, evaluated the signaling pathways involved in TGFß-dependent endothelial cells activation and the efficacy of anti-TGFß peptides in the control of MCF7-TGFß-dependent angiogenesis.ResultsTGFß over-expression induced in MCF7 several markers of the epithelial-to-mesenchymal transition. Conditioned-medium of TGFß-transfected MCF7 stimulated angiogenesis in vivo and in vitro by subsequent activation of SMAD2/3 and SMAD1/5 signaling in endothelial cells, as well as SMAD4 nuclear translocation, resulting in over-expression of the pro-angiogenic growth and differentiation factor-5 (GDF5). Inhibition or silencing of GDF5 in TGFß-stimulated EC resulted in impairment of GDF5 expression and of TGFß-dependent urokinase-plasminogen activator receptor (uPAR) overproduction, leading to angiogenesis impairment. Two different TGFß antagonist peptides inhibited all the angiogenesis-related properties elicited in EC by exogenous and conditionally-expressed TGFß in vivo and in vitro, including SMAD1/5 phosphorylation, SMAD4 nuclear translocation, GDF5 and uPAR overexpression. Antagonist peptides and anti-GDF5 antibodies efficiently inhibited in vitro and in vivo angiogenesis.ConclusionsTGFß produced by breast cancer cells induces in endothelial cells expression of GDF5, which in turn stimulates angiogenesis both in vitro and in vivo. Angiogenesis activation is rapid and the involved mechanism is totally opposed to the old and controversial dogma about the AKL5/ALK1 balance. The GDF-dependent pro-angiogenic effects of TGFß are controlled by anti-TGFß peptides and anti-GDF5 antibodies, providing a basis to develop targeted clinical studies.

""\\"Histone deacetylase inhibitors (HDACi) induce tumour cell cycle arrest and\\\\\\\/or apoptosis, and some of them are currently used in cancer therapy. Recently, we described a series of powerful HDACi characterized by a 1,4-benzodiazepine (BDZ) ring hybridized with a linear alkyl chain bearing a hydroxamate function as Zn(++) -chelating group. Here, we explored the anti-leukaemic properties of three novel hybrids, namely the chiral compounds (S)-2 and (R)-2, and their non-chiral analogue 4, which were first comparatively tested in promyelocytic NB4 cells. (S)-2 and partially 4- but not (R)-2 - caused G0\\\\\\\/G1 cell-cycle arrest by up-regulating cyclin G2 and p21 expression and down-regulating cyclin D2 expression, and also apoptosis as assessed by cell morphology and cytofluorimetric assay, histone H2AX phosphorylation and PARP cleavage. Notably, these events were partly prevented by an anti-oxidant. Moreover, novel HDACi prompted p53 and α-tubulin acetylation and, consistently, inhibited HDAC1 and 6 activity. The rank order of potency was (S)-2 > 4 > (R)-2, reflecting that of other biological assays and addressing (S)-2 as the most effective compound capable of triggering apoptosis in various acute myeloid leukaemia (AML) cell lines and blasts from patients with different AML subtypes. Importantly, (S)-2 was safe in mice (up to 150 mg\\\\\\\/kg\\\\\\\/week) as determined by liver, spleen, kidney and bone marrow histopathology; and displayed negligible affinity for peripheral\\\\\\\/central BDZ-receptors. Overall, the BDZ-hydroxamate (S)-2 showed to be a low-toxic HDACi with powerful anti-proliferative and pro-apototic activities towards different cultured and primary AML cells, and therefore of clinical interest to support conventional anti-leukaemic therapy.\\"""

Introduction: We collected the data relating to involuntary hospital treatment (IHT) in the University Psychiatric Ward at Novara Hospital between 1991 and 2002, and compared them with those relating to Piedmont and the whole of Italy. Methods: The data were collected from the ward medical records. Results: IHT was much more frequent among young male schizophrenics living with their families of origin. Most of the subjects were not working at the time of admission. There was a statistically significant correlation between male gender and the risk of being admitted for a period of less than 12 days. The risk of being admitted for more than 12 days significantly correlated with the province of birth and residence, as well as with a diagnosis of schizophrenic psychosis. Conclusions: Schizophrenia is the diagnosis that is most frequently associated with IHT.

The effect of amitriptyline on hypothalamic-pituitary-adrenocortical (HPA) axis activity was compared with that of fluvoxamine in 38 patients suffering from DMS-IV major depressive disorder. Basal plasma adrenocorticotropic hormone and cortisol levels were determined in the so-called "observation window" of an hour (08:00-09:00 h), and cortisol levels were determined again at 20:00 h. Clinical and biochemical assessments were performed before therapy (T0), at day 14 (T14), and at day 42 (T42) of the course of antidepressant treatment. At T0, neuroendocrine parameters did not differ in patients from those in controls, except for the ratio between cortisol levels at 20:00 h and the mean level of the "window" (ratio F20/F8), which was significantly higher, suggesting a dysregulation of the circadian pattern of cortisol. Although a decrease in the ratio F20/F8 was already apparent at T14 of both treatments, the repeated measures analysis of variance failed to demonstrate a significant variation with time (T0, T14, and T42) and with treatment (amitriptyline and fluvoxamine) for any hormonal measure. At T42, both treated groups showed a similar level of clinical improvement. Our results did not demonstrate any effect of antidepressant therapy on the cortisol circadian rhythm abnormality.

Background Suicidal behaviours are major public health concerns worldwide. They are associated with risk factors that vary with age and gender, occur in combination, and may change over time. The aim of our study was to investigate how frequently patients visiting a hospital emergency room (ER) require a psychiatric consultation for attempted suicide, and to outline the characteristics of this population. Methods Determinants of emergency room visits for psychiatric reasons were studied prospectively from 2008 to 2011 at the “Maggiore” Hospital in Novara. Results 280 out of 1888 patients requiring psychiatric consultation were referred to the ER because of suicide attempt. Suicide attempters were more often female. The rate of suicide attempters among Italian people was 14.2%, compared to 19.5% in foreigners. Subjects living with parents or own family and those having a permanent job had a higher frequency of suicide attempt. Suicide attempts were more frequent among patients with a history of psychiatric disorders; nonetheless, suicide attempts were more common among those who had not previously been hospitalized in a psychiatric ward or were not under the care of a psychiatrist. The multivariate analysis found that female gender was a risk factor for suicide attempt, while being in the colder months of the year and, surprisingly, unemployment were protective factors. Conclusions A better understanding of patients referring to the ER due to attempted suicide may allow the identification of at-risk subjects and the implementation of targeted treatment approaches.