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Uveal melanoma (UM) is a rare ocular tumor that may lead to deadly metastases in 50% of patients. A disintegrin and metalloproteinase (ADAM)10, ADAM17, and the HGF-receptor c-Met support invasiveness in different tumors. Here, we report that high ADAM10, MET, and, to a lesser extent, ADAM17 gene expression correlates with poor progression-free survival in UM patients (hazard ratio 2.7, 2.6, and 1.9, respectively). About 60% of primary UM expresses c-Met and/or ADAM10 proteins. Four UM cell lines display high levels of ADAM10 and ADAM17, which constitutively cleave c-Met, inducing the release of soluble c-Met. ADAM10/17 pharmacological inhibition or gene silencing reduces c-Met shedding, but has limited impact on surface c-Met, which is overexpressed. Importantly, ADAM10 silencing inhibits UM cell invasion driven by FCS or HGF, while ADAM17 silencing has a limited effect. Altogether our data indicate that ADAM10 has a pro-invasive role and may contribute to UM progression.

Observations on the role of ovarian hormones in breast cancer growth, as well as interest in contraception, stimulated research into the biology of estrogens. The identification of the classical receptors ERα and ERβ and the transmembrane receptor GPER and the resolution of the structure of the ligand bound to its receptor established the principal molecular mechanisms of estrogen action. The presence of estrogen-like compounds in many plants used in traditional medicine or ingested as food ingredients, phytoestrogens, as well as the estrogenic activities of many industrial pollutants and pesticides, xenoestrogens, have prompted investigations into their role in human health. Phyto- and xenoestrogens bind to the estrogen receptors with a lower affinity than the endogenous estrogens and can compete or substitute the hormone. Xenoestrogens, which accumulate in the body throughout life, are believed to increase breast cancer risk, especially in cases of prenatal and prepuberal exposure whereas the role of phytoestrogens is still a matter of debate. At present, the application of phytoestrogens appears to be limited to the treatment of post-menopausal symptoms in women where the production of endogenous estrogens has ceased. In this review we discuss chemistry, structure and classification, estrogen signaling and the consequences of the interactions of estrogens, phytoestrogens and xenoestrogens with their receptors, the complex interactions of endogenous and exogenous ligands, the evaluation of the health risks related to xenoestrogens, and the perspectives toward the synthesis of potent third generation selective estrogen receptor modulators (SERMs).

Several substances widely dispersed in the environment including hormones, industrial by-products and pollutants exert hormone like activity affecting steroid-responsive physiological systems. These compounds, named endocrine disruptors, are suspected to affect the mammalian reproductive system. However it is still unclear whether these substances are able to elicit estrogen like activity at the low concentrations encountered in the environment. Here we compare the effects of the endocrine disruptor nonylphenol with the effects elicited by 17-β -estradiol on gene transcription in the human breast cancer cell line MCF7. The correlation of the nonylphenol induced gene expression alterations with a reference profile of estradiol treated cells shows that nonylphenol at a concentration of 100 nM exerts a significant effect on estrogen responsive gene transcription in MCF7 cells. Most of the genes regulated by 17-β -estradiol respond to the nonylphenol in the same direction though to a much lesser extent. Molecular modeling of the potential interaction of nonylphenol with the estrogen receptor α shows that nonylphenol is likely to bind to the estrogen receptor α.

Uveal melanoma is the most common primary tumour of the eye with an annual incidence of approximately two cases per million in southern European countries to eight cases in northern European countries (Virgili et al, 2007). Incidence increases with latitude in a highly significant manner (Virgili et al, 2007). Whether this association can be attributed to the exposure to sunlight of variable intensity or not, remains a matter of discussion (Gallagher et al, 1985; Pane and Hirst, 2000; Guenel et al, 2001; Singh et al, 2004). Uveal melanoma shows a mutation pattern that is clearly distinct from cutaneous (for recent reviews, see Coupland et al, 2013; Harbour, 2013; Zeschnigk and Lohmann, 2013), mucosal (Furney et al, 2013) and conjunctival melanomas (Griewank et al, 2013c). The mutations typically encountered in cutaneous and conjunctival melanomas, BRAF and NRAS, are rare in uveal melanomas that are characterised by mutations of the G-proteins GNAQ and GNA11 occurring in mutual exclusive manner in ∼85% of the cases (Zeschnigk and Lohmann, 2013). The mutation pattern observed by exome sequencing in cutaneous melanoma is clearly consistent with an aetiological role of sunlight exposure (Hodis et al, 2012). Cytidine-to-thymidine transition frequency has not been addressed by the two exome sequencing studies of uveal melanoma (Harbour et al, 2013; Martin et al, 2013) but the complete lack of overlap among the driver mutations in the two pathologies is consistent with a different aetiology. Recently, three mutations in the promoter of the telomerase reverse transcriptase (TERT) needed for telomere maintenance in cancer cells, close to the transcriptional start site, have been described for sporadic (Huang et al, 2013) and familiar (Horn et al, 2013) forms of cutaneous melanoma. The familiar cases all derived from one family and showed a mutation different from those detected in sporadic melanomas and melanoma cell lines (Huang et al, 2013). Two mutations were identified in sporadic cases where they occurred in a mutually exclusive manner in over 70% of cases. The same mutations were also present, though less frequently, in other tumours and tumour-derived cell lines. The mutations consisted in cytidine-to-thymidine transitions at a dipyrimidine motif consistent with ultraviolet (UV) light-induced damage. All three mutations created novel binding sites for the transcription factor E-twenty-six (ETS) in the TERT promoter within 100 bp upstream of the transcription start site (TSS) in sporadic cases and closer to the TTS (−57 bp) in the familiar cases. Reporter assays showed the functionality of the new binding sites observed in sporadic cases (Huang et al, 2013). The mutation is highly penetrant in the melanoma family analysed and it is linked to early onset (mean 34 years) and short survival (mean 3.5 years in 8 patients who died from the disease of a total of 11 cases). It is therefore probable that the mutation strongly contributes to the development of aggressively growing cutaneous melanoma. Griewank et al (2013a) have addressed whether the same mutations are present also in ocular melanomas reporting an incidence of 32% in conjunctival melanomas and the absence from uveal (ciliary body or choroid) melanomas (n=47). In the present study, we describe the analysis of TERT promoter mutations in a series of 50 uveal melanomas. A mutation was detected in a single case that also carried mutations in GNA11 and EIF1AX, typical for this disease and absent from conjunctival and cutaneous melanomas. The tumour showed disomy of chromosome 3 and had wild-type sequences of GNAQ, BAP1 and SF3B1 which frequently show mutations in uveal melanoma.

Biomarkers are important for early detection of cancer, prognosis, response prediction, and detection of residual or relapsing disease. Special attention has been given to diagnostic markers for prostate cancer since it is thought that early detection and surgery might reduce prostate cancer-specific mortality. The use of prostate-specific antigen, PSA (KLK3), has been debated on the base of cohort studies that show that its use in preventive screenings only marginally influences mortality from prostate cancer. Many groups have identified alternative or additional markers, among which PCA3, in order to detect early prostate cancer through screening, to distinguish potentially lethal from indolent prostate cancers, and to guide the treatment decision. The large number of markers proposed has led us to the present study in which we analyze these indicators for their diagnostic and prognostic potential using publicly available genomic data. We identified 380 markers from literature analysis on 20,000 articles on prostate cancer markers. The most interesting ones appeared to be claudin 3 (CLDN3) and alpha-methysacyl-CoA racemase highly expressed in prostate cancer and filamin C (FLNC) and keratin 5 with highest expression in normal prostate tissue. None of the markers proposed can compete with PSA for tissue specificity. The indicators proposed generally show a great variability of expression in normal and tumor tissue or are expressed at similar levels in other tissues. Those proposed as prognostic markers distinguish cases with marginally different risk of progression and appear to have a clinically limited use. We used data sets sampling 152 prostate tissues, data sets with 281 prostate cancers analyzed by microarray analysis and a study of integrated genomics on 218 cases to develop a multigene score. A multivariate model that combines several indicators increases the discrimination power but does not add impressively to the information obtained from Gleason scoring. This analysis of 10 years of marker research suggests that diagnostic and prognostic testing is more difficult in prostate cancer than in other neoplasms and that we must continue to search for better candidates. Electronic supplementary material The online version of this article (doi:10.1007/s10555-013-9470-4) contains supplementary material, which is available to authorized users.

The anti-hyperglycaemic drug metformin has important anticancer properties as shown by the direct inhibition of cancer cells proliferation. Tumor cells avidly use glucose as a source for energy production and cell building blocks. Critical to this phenotype is the production of glucose-6-phosphate (G6P), catalysed by hexokinases (HK) I and II, whose role in glucose retention and metabolism is highly advantageous for cell survival and proliferation. Here we show that metformin impairs the enzymatic function of HKI and II in Calu-1 cells. This inhibition virtually abolishes cell glucose uptake and phosphorylation as documented by the reduced entrapment of (18)F-fluorodeoxyglucose. In-silico models indicate that this action is due to metformin capability to mimic G6P features by steadily binding its pocket in HKII. The impairment of this energy source results in mitochondrial depolarization and subsequent cell death. These results could represent a starting point to open effective strategies in cancer prevention and treatment.

Breast cancer is the most frequent female cancer and still one of the major causes of death although early diagnosis and improved therapies have had a great impact on survival after breast cancer diagnosis. However, there are still many unresolved problems in breast cancer such as the fraction of breast cancers that do not respond to current therapies and considerable overtreatment due to imperfect prognostication. The application of genomics to breast cancer has led to the identification of clinically relevant molecular subtypes, especially the distinction of luminal A and luminal B subtypes within the class of hormone receptor positive cancers. Many prognostic signatures have been developed and two of them are being applied in oncologic decision making yet their utility most likely does not go beyond the distinction of luminal A and B subtypes that show a highly different proliferative potential. Integration of copy number variation has identified even more subclasses with distinct clinical characteristics. Genome wide association studies have identified many single nucleotide polymorphisms that are associated with breast cancer risk and several of them resist in validation studies. Their application for the design of risk based preventive strategies has been proposed. Next generation sequencing shows a wide variation of driver mutations in breast cancer, most of them within interrelated signaling pathways. Several genes such as TP53 or PIK3CA show frequent mutations but many mutations are almost private. Sequencing also identified several actionable mutations, among which those that occur in genes more frequently involved in other cancers that could indicate specific treatments. Better prognostication and response prediction by means of genomic analyses and mutation screening will almost certainly contribute to the improvement of therapy and to the reduction of unnecessary toxicity. Breast cancer genomics has also led to a conceptual shift in our understanding of the process of metastasis that seems to be determined from very early stages of the disease although additional mutations occur at later stages.

Survival after diagnosis of laryngeal cancer has not improved over the last 20 years. Selection of patients for radio- and chemotherapy or surgery or follow-up strategies based on a prognostic classifier could improve survival without unduly extending radical surgery. We performed microarray gene expression analysis and developed a four-gene classifier for laryngeal cancer using Prediction Analysis of Microarray and leave-one-out cross validation. A four-gene classifier containing the non-coding gene H19, the histone HIST1H3F and the two small nucleolar RNAs, SNORA16A and SNORD14C was developed that assigns cases to low and high risk classes. The high risk class has a relative risk of 6.5 (CI = 1.817–23.258, Fisher exact test p

Este trabalho é dedicado à modelagem por elementos finitos de estruturas compostas laminadas incorporando materiais viscoelásticos para o controle passivo de vibrações e ruídos. Neste contexto, foi estudada a dependência da capacidade de amortecimento dos materiais viscoelásticos com relação à frequência de excitação e temperatura e à incorporação do amortecimento viscoelástico em modelos de elementos finitos de sistemas estruturais. Para a modelagem de estruturas compostas laminadas foi empregada a Teoria da Deformação Cisalhante de Ordem Superior combinada com um elemento de placa plana retangular do tipo Serendipity contendo oito nós e onze graus de liberdade por nó. O amortecimento viscoelástico é representado pelo modelo do módulo complexo associado ao conceito de fator de deslocamento e frequência reduzida de acordo com o Princípio da Superposição Frequência-Temperatura. Além disso, foi implementado um método de redução de sistemas amortecidos viscoelasticamente via utilização de uma base de redução constante (independente da frequência e da temperatura) com o objetivo de reduzir o tempo computacional requerido para o cálculo das respostas dinâmicas dos sistemas amortecidos. Para avaliar a influência dos parâmetros de projeto nas respostas dinâmicas, é apresentada uma formulação baseada em derivadas de primeira ordem para a análise de sensibilidade das funções de resposta em frequência em relação a um conjunto de parâmetros de projeto pré-definidos e que foram fatorados das matrizes elementares de elementos finitos via procedimento de parametrização do modelo. Toda a modelagem matemática foi implementada computacionalmente utilizando o ambiente de programação MATLAB®, e os resultados obtidos permitiram não só avaliar o desempenho dos materiais viscoelásticos em termos da atenuação dos níveis de vibração, mas também, ilustrar os procedimentos de modelagem e incorporação do amortecimento viscoelástico em modelos de elementos finitos de estruturas compostas.

Background Obesity and the insulin resistance syndrome are risk factors for breast cancer and might also affect breast cancer progression. The anti-diabetic drug Metformin (METF) reduces the breast cancer risk in diabetic women. Insulin like growth factor 1 (IGF1) and insulin are involved in breast cancer tumorigenesis and progression. We tested the effect of METF on the IGF1/insulin pathway and its involvement in breast cancer progression. Methods We developed a prognostic signature based on IGF1/insulin pathway genes using the Stockholm breast cancer microarray dataset of 149 cases for training and primary validation and the Uppsala dataset of 249 for external validation. The effect of METF on the prognostic gene set identified was tested in vitro on a panel of breast cancer cell lines. METF effects on proliferation and glucose metabolism were analyzed in vitro and in vivo. The insulin receptor substrate 2 (IRS2) was silenced by transfection with shRNA-lentiviral vectors. Xenograft growth, in the presence and absence of METF, was studied and 18FDG-uptake was measured in vitro and in vivo. Results A 15-gene signature (Insulin sensitivity score, ISS) was developed and predicted breast cancer metastasis with an accuracy similar to the Recurrence Score. ISS genes were expressed at variable levels in a breast cancer cell line panel and showed variable responsiveness to METF. The high expression correlation among the ISS genes observed in untreated breast cancer cell lines was lost upon treatment with METF. METF reduced breast cancer cell growth in vitro with IC50 values ranging from 1mM to 25mM. Growth of MDA-MB-231 cells and hyper-invasive subpopulations derived therefrom was reduced in vivo by oral administration of METF to xenografted nude mice. Response to METF in terms of IC50 values correlated with basal expression of the 15 ISS genes with the strongest inverse correlation observed for IRS2. Stable silencing of IRS2 reduced the MDA-231 cell responsiveness to METF in vitro. Discussion METF acts on the insulin/IGF1 axis by disturbing a network of breast cancer progression related genes and appears to depend in its action on the expression of IRS2 that inversely correlates with the sensitivity of cell lines to the drug. The disruption of the ISS gene network is expected to correlate with an effect on breast cancer growth and progression and in fact, mouse xenografts show reduced growth upon treatment with METF. IRS2 appears to be a major mediator of METF effects. Citation Format: Alessia I. Esposito, Adriana Amaro, Giovanna Angelini, Laura Emionite, Alessandra Gennari, Stefano Indraccolo, Davide Maggi, Cecilia Marini, Barabara Salani, Gianmario Sambuceti, Maria Pia Sormani, Ulrich Pfeffer. Metformin affects breast cancer cell growth and disturbs an IGF1/insulin related gene network that correlates with breast cancer progression. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1182. doi:10.1158/1538-7445.AM2015-1182

Uveal melanoma is the most frequent primary tumor of the eye. It is molecularly clearly distinct from cutaneous melanoma and shows a different pattern of driver mutations. The influence of sunlight UV-exposure on the etiology of uveal melanoma is matter of debate. The recent identification of driver mutations in the promoter of the telomerase reverse transcriptase (TERT) gene with UV induced cytidine-to-thymidine transitions in cutaneous melanoma prompted us to investigate whether these mutations also occur in uveal melanoma. We also analyzed the frequency of other recently discovered uveal melanoma specific mutations in the genes GNAQ, GNA11, BAP1, SF3B1, and EIFAX1, and we analyzed the association of these mutation with histopathological and clinical features including progression free survival. Finally, we analyzed TERT expression in these tumors. The mutation frequencies observed confirmed the prevalence of GNAQ mutations in cases with disomy of chromosome 3 that have a better prognosis whereas GNA11 was more frequent in monosomic cases. Together these two mutations account for more than 85% of the cases. BAP1 mutations are associated with chromosome 3 monosomy but not with progression free survival. SF3B1 and EIF1AX mutations occur in 10.3 and 22.5% of uveal melanomas, in disomic cases only. We detected a TERT mutation in only one case of a 57-year old white male with clinical and histo-pathological features typical for uveal melanoma. The tumor showed mutations in GNA11 and EIF1AX. No mutations were detected in GNAQ, BAP1, and SF3B1. Both copies of chromosome 3 were retained. Several tumors among which the one carrying the TERT promoter mutation showed elevated TERT expression. These data indicate that TERT mutations are rare in uveal melanoma, consistent with a reduced etiological influence of sunlight. No conclusion can be drawn on the potential influence of TERT mutations on tumor progression. Citation Format: Adriana Agnese AMARO, Marina Gualco, Maria Dono, Ulrich Pfeffer, Irena Maric, Carlo Mosci. TERT promoter mutations are rare in uveal melanoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3420. doi:10.1158/1538-7445.AM2014-3420

The acquisition of an invasive phenotype is a pre-requisite for metastasis. We set out to develop cellular systems that can mirror transient and stbale molecular alterations that confer an invasive phenotype to breast cancer cells. We observed that it is possible to isolate invasive subpopulations from moderately invasive cancer cell lines. Enrichment of invasive sub-populations of MDA-MB-231 breast cancer cells in three successive preparative invasion assays in Matrigel covered Boyden chambers yielded a highly invasive cell line. Prolonged cultivation of these cells did not abolish the invasive phenotype although not all phenotypic changes acquired during selection are maintained by long term cultures. Genetic analyses of these cells by cytogenetics and array based comparative genome hybridization revealed many genetic alterations including increased ploidy. The flow cytometric DNA Index (DI) changes from 1.28 to 2.28. Cells with DI 2.28 constitute 1.4% of the parental cell line. Whole genome SNP analysis shows that the two populations are genetically related excluding any cell contamination. The invasive cells proliferate and undergo apoptosis similar to the parental cells. Commitment to apoptosis is increased since invasive cells respond more strongly to curcumin or peroxide induced apoptosis. Invasive cells show relative resistance to the cytotoxic, alkylating agent Doxorubicin and increased sensitivity to the anti-mitotic drugs Vincristine and Taxol. Increased resistance to the topoisomerase II inhibitor Mitoxantrone is observed only transiently in invasive cells and lost in long term cultures. Similarly, the chemokines CXCL1 and -2 are transiently upregulated. Response to the anti-diabetic drug Metformin showed a reduced sensibility for invasive cells in terms of growth inhibition (IC50). Gene expression profiling shows complex alterations in gene expression. Many of the genes that are differentially expressed in highly versus moderately invasive cells are differentially expressed in human breast cancer cases with and without distant metastasis and correlate with disease free survival. The invasive phenotype is not related to stem cell features nor to epithelial mesenchymal transition. These cells constitute a novel model for tumor progression. Tumorigenicity and metastatic potential in vivo are currently being tested. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 496. doi:1538-7445.AM2012-496

// Giovanna Bianchi 1,* , Roberto Martella 1,* , Silvia Ravera 2 , Cecilia Marini 3 , Selene Capitanio 4 , Annamaria Orengo 4 , Laura Emionite 5 , Chiara Lavarello 6 , Adriana Amaro 7 , Andrea Petretto 6 , Ulrich Pfeffer 7 , Gianmario Sambuceti 4 , Vito Pistoia 1 , Lizzia Raffaghello 1,** and Valter D. Longo 8,9,10,** 1 Laboratorio di Oncologia Istituto G. Gaslini, Genoa, Italy 2 Department of Pharmacy, University of Genoa, Genova, Italy 3 CNR Institute of Bioimages and Molecular Physiology, Milan, Section of Genoa, Genoa, Italy 4 Nuclear Medicine Unit, Department of Health Sciences, University of Genoa and IRCCS AOU San Martino - IST Istituto Nazionale per la Ricerca sul Cancro, Genoa, Italy 5 Animal facility, IRCCS AOU San Martino - IST Istituto Nazionale per la Ricerca sul Cancro, Genoa, Italy 6 Core Facility, Istituto G. Gaslini, Genoa, Italy 7 Functional Genomics, IRCCS AOU San Martino - IST Istituto Nazionale per la Ricerca sul Cancro, Genoa, Italy 8 Longevity Institute, School of Gerontology, Department of Biological Sciences, University of Southern California, Los Angeles, CA, USA 9 Eli and Edythe Broad Center for Regenerative Medicine and Stem Cell Research at USC, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA 10 IFOM, FIRC Institute of Molecular Oncology, Milan, Italy * These authors have contributed equally as first authors ** These authors have contributed equally as last authors Correspondence to: Valter D. Longo, email: // Keywords : fasting, Warburg effect, colon cancer, oxidative phosphorylation, glucose uptake Received : March 04, 2015 Accepted : March 11, 2015 Published : March 30, 2015 Abstract Tumor chemoresistance is associated with high aerobic glycolysis rates and reduced oxidative phosphorylation, a phenomenon called “Warburg effect” whose reversal could impair the ability of a wide range of cancer cells to survive in the presence or absence of chemotherapy. In previous studies, Short-term-starvation (STS) was shown to protect normal cells and organs but to sensitize different cancer cell types to chemotherapy but the mechanisms responsible for these effects are poorly understood. We tested the cytotoxicity of Oxaliplatin (OXP) combined with a 48hour STS on the progression of CT26 colorectal tumors. STS potentiated the effects of OXP on the suppression of colon carcinoma growth and glucose uptake in both in vitro and in vivo models. In CT26 cells, STS down-regulated aerobic glycolysis, and glutaminolysis, while increasing oxidative phosphorylation. The STS-dependent increase in both Complex I and Complex II-dependent O 2 consumption was associated with increased oxidative stress and reduced ATP synthesis. Chemotherapy caused additional toxicity, which was associated with increased succinate/Complex II-dependent O 2 consumption, elevated oxidative stress and apoptosis . These findings indicate that the glucose and amino acid deficiency conditions imposed by STS promote an anti-Warburg effect characterized by increased oxygen consumption but failure to generate ATP, resulting in oxidative damage and apoptosis.

Melanoma cells with acquired resistance to dabrafenib display changes in miRNA expression pattern and respond to this drug with an increase of invasiveness, which is abrogated by inhibition of NFB or the PI3K/mTOR signalling pathway Simona Caporali, Ester Alvino, Adriana Amaro, Pedro Miguel Lacal, Lauretta Levati, Maria Grazia Atzori, Gian Carlo Antonini Cappellini, Federica Ruffini, Enzo Bonmassar, Ulrich Pfeffer, Stefania D’Atri