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[Abstract] Human high-mobility group-B (HMGB) proteins regulate gene expression in prostate cancer (PCa), a leading cause of oncological death in men. Their role in aggressive PCa cancers, which do not respond to hormonal treatment, was analyzed. The effects of HMGB1 and HMGB2 silencing upon the expression of genes previously related to PCa were studied in the PCa cell line PC-3 (selected as a small cell neuroendocrine carcinoma, SCNC, PCa model not responding to hormonal treatment). A total of 72% of genes analyzed, using pre-designed primer panels, were affected. HMGB1 behaved mostly as a repressor, but HMGB2 as an activator. Changes in SERPINE1, CDK1, ZWINT, and FN1 expression were validated using qRT-PCR after HMGB1 silencing or overexpression in PC-3 and LNCaP (selected as an adenocarcinoma model of PCa responding to hormonal treatment) cell lines. Similarly, the regulatory role of HMGB2 upon SERPINE1, ZWINT, FN1, IGFPB3, and TYMS expression was validated, finding differences between cell lines. The correlation between the expression of HMGB1, HMGB2, and their targets was analyzed in PCa patient samples and also in PCa subgroups, classified as neuroendocrine positive or negative, in public databases. These results allow a better understanding of the role of HMGB proteins in PCa and contribute to find specific biomarkers for aggressive PCa.

[Abstract] Ovarian cancer is one of the most lethal gynecological malignancies worldwide due to the lack of early diagnostic methods. In this regard, HMGB family members, HMGB1 and HMGB2, as well as long non-coding RNAs (lncRNAs), participate in the malignant transformation by regulating gene expression and, therefore, their study represents an opportunity to progress towards a better understanding of the disease and the development of novel clinical strategies. The main objective of this work was to identify lncRNAs in epithelial ovarian cancer that are deregulated and/or associated with HMGB1 and/or HMGB2 because of their potential value as novel diagnostic biomarkers and therapeutic targets. The determination of new RNA interactions of these proteins would allow a better understanding of their mechanism of action and their role in ovarian cancer pathology. We performed a meta-analysis on transcriptomic profiling expression data from epithelial ovarian cancer patients that are publicly available. By this approach, we identified several lncRNAs previously unrelated to epithelial ovarian cancer, with diagnostic and prognostic values, and associated with metastasis, chemoresistance, or histological subtype. Then, the RNA interactome of HMGB1 and HMGB2 was determined by applying in silico binding predictions, as well as two immunoprecipitation-based techniques, RIP-Seq and eCLIP in a high-grade serous epithelial ovarian cancer cell line, PEO1. The identified lncRNAs and messenger RNAs (mRNAs) are related to previous information present in scientific literature and gene ontologies regarding biological processes, respectively. Finally, HMGB1 and HMGB2 silencing was carried out using an siRNA-based approach and their effects on PEO1 transcriptome were analyzed. Their role in signaling pathways and biological processes in addition to the HMGB-regulated lncRNAs were assessed., [Resumen] El cáncer de ovario es uno de los cánceres ginecológicos más letales en todo el mundo debido a la falta de métodos de diagnóstico precoz. En este sentido, los miembros de la familia High Mobility Group Box (HMGB), HMGB1 y HMGB2, así como los ARNs largos no codificantes (lncRNAs), participan en el proceso de malignización regulando la expresión génica y, por tanto, su estudio representa una oportunidad para avanzar hacia una mejor comprensión de la enfermedad y el desarrollo de nuevas estrategias clínicas. El objetivo principal de este trabajo consistió en identificar lncRNAs en cáncer de ovario epitelial que estén desregulados y/o asociados a HMGB1 y/o HMGB2, debido a su potencial valor como nuevos biomarcadores diagnósticos y dianas terapéuticas. La determinación de nuevas interacciones de estas proteínas con ARNs permitirá una mejor comprensión de su mecanismo de acción y su papel en la patología del cáncer de ovario. Realizamos un metaanálisis de datos disponibles públicamente de transcriptomas de pacientes con cáncer de ovario epitelial. Mediante esta aproximación, identificamos varios lncRNAs que no estaban previamente relacionados con el cáncer de ovario epitelial, y que tienen valor diagnóstico y pronóstico, o están asociados a metástasis, resistencia a quimioterapia o subtipo histológico. A continuación, se determinó el interactoma de ARN para HMGB1 y HMGB2 aplicando predicciones de unión in silico, así como dos técnicas basadas en inmunoprecipitación, RIP-Seq y eCLIP en la línea celular de cáncer de ovario epitelial seroso de alto grado, PEO1. Los lncRNAs y ARN mensajeros (ARNm) identificados se relacionaron con información previa presente en la bibliografía científica y ontologías génicas relativas a procesos biológicos, respectivamente. Por último, se llevó a cabo el silenciamiento de HMGB1 y HMGB2 utilizando una aproximación basada en siRNA y se estudiaron sus efectos en el transcriptoma de PEO1. Se evaluó su papel en vías de señalización y pro, [Resumo] O cancro de ovario é un dos cancros xinecolóxicos máis letais en todo o mundo debido á falta de métodos de diagnóstico precoz. Neste senso, os membros da familia High Mobility Group Box (HMGB), HMGB1 e HMGB2, así como os ARNs longos non codificantes (lncRNAs) participan no proceso de malignización regulando a expresión xénica e, por tanto, o seu estudo representa unha oportunidade para avanzar cara a unha mellor comprensión da enfermidade e o desenvolvemento de novas estratexias clínicas. O obxectivo principal deste traballo consistiu en identificar lncRNAs en cancro de ovario epitelial que estean deregulados e/ou asociados a HMGB1 e/o HMGB2, debido ao seu potencial valor como novos biomarcadores diagnósticos e dianas terapéuticas. A determinación de novas interaccións destas proteínas con ARNs permitirá unha mellor comprensión do seu mecanismo de acción e o seu papel na patoloxía do cancro de ovario. Realizamos unha meta-análise de datos dispoñibles publicamente de transcriptomas de pacientes con cancro de ovario epitelial. Mediante esta aproximación, identificamos varios lncRNAs que non estaban previamente relacionados co cancro de ovario epitelial, e que teñen valor diagnóstico e prognóstico, ou están asociados a metástase, resistencia á quimioterapia ou subtipo histolóxico. A continuación, determinouse o interactoma de ARN para HMGB1 e HMGB2 aplicando predicións de unión in silico, así como dúas técnicas baseadas en inmunoprecipitación, RIP-Seq e eCLIP na liña celular de cancro de ovario epitelial seroso de alto grao, PEO1. Os lncRNAs e ARN mensaxeiros (ARNm) identificados relacionáronse con información previa presente na bibliografía científica e ontoloxías xénicas relativas a procesos biolóxicos, respectivamente. Por último, levouse a cabo o silenciamento de HMGB1 e HMGB2 utilizando unha aproximación baseada en siRNA e se estudaron os seus efectos no transcriptoma de PEO1. Avaliouse o seu papel nas vías de sinalización e os procesos biolóxicos, así co

[Abstract] Ovarian cancer is one of the most lethal gynecological malignancies worldwide due to the lack of early diagnostic methods. In this regard, HMGB family members, HMGB1 and HMGB2, as well as long non-coding RNAs (lncRNAs), participate in the malignant transformation by regulating gene expression and, therefore, their study represents an opportunity to progress towards a better understanding of the disease and the development of novel clinical strategies. The main objective of this work was to identify lncRNAs in epithelial ovarian cancer that are deregulated and/or associated with HMGB1 and/or HMGB2 because of their potential value as novel diagnostic biomarkers and therapeutic targets. The determination of new RNA interactions of these proteins would allow a better understanding of their mechanism of action and their role in ovarian cancer pathology. We performed a meta-analysis on transcriptomic profiling expression data from epithelial ovarian cancer patients that are publicly available. By this approach, we identified several lncRNAs previously unrelated to epithelial ovarian cancer, with diagnostic and prognostic values, and associated with metastasis, chemoresistance, or histological subtype. Then, the RNA interactome of HMGB1 and HMGB2 was determined by applying in silico binding predictions, as well as two immunoprecipitation-based techniques, RIP-Seq and eCLIP in a high-grade serous epithelial ovarian cancer cell line, PEO1. The identified lncRNAs and messenger RNAs (mRNAs) are related to previous information present in scientific literature and gene ontologies regarding biological processes, respectively. Finally, HMGB1 and HMGB2 silencing was carried out using an siRNA-based approach and their effects on PEO1 transcriptome were analyzed. Their role in signaling pathways and biological processes in addition to the HMGB-regulated lncRNAs were assessed., [Resumen] El cáncer de ovario es uno de los cánceres ginecológicos más letales en todo el mundo debido a la falta de métodos de diagnóstico precoz. En este sentido, los miembros de la familia High Mobility Group Box (HMGB), HMGB1 y HMGB2, así como los ARNs largos no codificantes (lncRNAs), participan en el proceso de malignización regulando la expresión génica y, por tanto, su estudio representa una oportunidad para avanzar hacia una mejor comprensión de la enfermedad y el desarrollo de nuevas estrategias clínicas. El objetivo principal de este trabajo consistió en identificar lncRNAs en cáncer de ovario epitelial que estén desregulados y/o asociados a HMGB1 y/o HMGB2, debido a su potencial valor como nuevos biomarcadores diagnósticos y dianas terapéuticas. La determinación de nuevas interacciones de estas proteínas con ARNs permitirá una mejor comprensión de su mecanismo de acción y su papel en la patología del cáncer de ovario. Realizamos un metaanálisis de datos disponibles públicamente de transcriptomas de pacientes con cáncer de ovario epitelial. Mediante esta aproximación, identificamos varios lncRNAs que no estaban previamente relacionados con el cáncer de ovario epitelial, y que tienen valor diagnóstico y pronóstico, o están asociados a metástasis, resistencia a quimioterapia o subtipo histológico. A continuación, se determinó el interactoma de ARN para HMGB1 y HMGB2 aplicando predicciones de unión in silico, así como dos técnicas basadas en inmunoprecipitación, RIP-Seq y eCLIP en la línea celular de cáncer de ovario epitelial seroso de alto grado, PEO1. Los lncRNAs y ARN mensajeros (ARNm) identificados se relacionaron con información previa presente en la bibliografía científica y ontologías génicas relativas a procesos biológicos, respectivamente. Por último, se llevó a cabo el silenciamiento de HMGB1 y HMGB2 utilizando una aproximación basada en siRNA y se estudiaron sus efectos en el transcriptoma de PEO1. Se evaluó su papel en vías de señalización y pro, [Resumo] O cancro de ovario é un dos cancros xinecolóxicos máis letais en todo o mundo debido á falta de métodos de diagnóstico precoz. Neste senso, os membros da familia High Mobility Group Box (HMGB), HMGB1 e HMGB2, así como os ARNs longos non codificantes (lncRNAs) participan no proceso de malignización regulando a expresión xénica e, por tanto, o seu estudo representa unha oportunidade para avanzar cara a unha mellor comprensión da enfermidade e o desenvolvemento de novas estratexias clínicas. O obxectivo principal deste traballo consistiu en identificar lncRNAs en cancro de ovario epitelial que estean deregulados e/ou asociados a HMGB1 e/o HMGB2, debido ao seu potencial valor como novos biomarcadores diagnósticos e dianas terapéuticas. A determinación de novas interaccións destas proteínas con ARNs permitirá unha mellor comprensión do seu mecanismo de acción e o seu papel na patoloxía do cancro de ovario. Realizamos unha meta-análise de datos dispoñibles publicamente de transcriptomas de pacientes con cancro de ovario epitelial. Mediante esta aproximación, identificamos varios lncRNAs que non estaban previamente relacionados co cancro de ovario epitelial, e que teñen valor diagnóstico e prognóstico, ou están asociados a metástase, resistencia á quimioterapia ou subtipo histolóxico. A continuación, determinouse o interactoma de ARN para HMGB1 e HMGB2 aplicando predicións de unión in silico, así como dúas técnicas baseadas en inmunoprecipitación, RIP-Seq e eCLIP na liña celular de cancro de ovario epitelial seroso de alto grao, PEO1. Os lncRNAs e ARN mensaxeiros (ARNm) identificados relacionáronse con información previa presente na bibliografía científica e ontoloxías xénicas relativas a procesos biolóxicos, respectivamente. Por último, levouse a cabo o silenciamento de HMGB1 e HMGB2 utilizando unha aproximación baseada en siRNA e se estudaron os seus efectos no transcriptoma de PEO1. Avaliouse o seu papel nas vías de sinalización e os procesos biolóxicos, así co

[Abstract] Ruthenium(II) complexes are currently considered attractive alternatives to the widely used platinum-based drugs. We present herein the synthesis and characterization of half-sandwich ruthenium compounds formulated as [Ru(p-cymene)(L)Cl][CF3SO3] (L = 1,1-bis(methylenediphenylphosphano)ethylene, 1; L = 1,1-bis(diphenylphosphano)ethylene, 2), which were characterized by elemental analysis, mass spectrometry, 1H and 31P{1H} NMR, UV–vis and IR spectroscopy, conductivity measurements and cyclic voltammetry. The molecular structures for both complexes were determined by single-crystal X-ray diffraction. Their cytotoxic activity was evaluated using the MTT assay against human tumor cells, namely ovarian (A2780) and breast (MCF7 and MDA-MB-231). Both complexes were active against breast adenocarcinoma cells, with complex 1 exhibiting a quite remarkable cytotoxicity in the submicromolar range. Interestingly, at concentrations equivalent to the IC50 values in the MCF7 cancer cells, complexes 1 and 2 presented lower cytotoxicity in normal human primary fibroblasts. The antiproliferative effects of 1 and 2 in MCF7 cells might be associated with the induction of reactive oxygen species (ROS), leading to a combined cell death mechanism via apoptosis and autophagy. Despite the fact that in vitro a partial intercalation between complexes and DNA was observed, no MCF7 cell cycle delay or arrest was observed, indicating that DNA might not be a direct target. Complexes 1 and 2 both exhibited a moderate to strong interaction with human serum albumin, suggesting that protein targets may be involved in their mode of action. Their acute toxicity was evaluated in the zebrafish model. Complex 1 (the most toxic of the two) exhibited a lethal toxicity LC50 value about 1 order of magnitude higher than any IC50 concentrations found for the cancer cell models used, highlighting its therapeutic relevance as a drug candidate in cancer chemotherapy.

[Abstract] Ovarian cancer is one of the most lethal gynecological malignancies worldwide because it tends to be detected late, when the disease has already spread, and prognosis is poor. In this review we aim to highlight the importance of long non-coding RNAs (lncRNAs) in diagnosis, prognosis and treatment choice, to make progress towards increasingly personalized medicine in this malignancy. We review the effects of lncRNAs associated with ovarian cancer in the context of cancer hallmarks. We also discuss the molecular mechanisms by which lncRNAs become involved in cellular physiology; the onset, development and progression of ovarian cancer; and lncRNAs’ regulatory mechanisms at the transcriptional, post-transcriptional and post-translational stages of gene expression. Finally, we compile a series of online resources useful for the study of lncRNAs, especially in the context of ovarian cancer. Future work required in the field is also discussed along with some concluding remarks.

[Abstract] High mobility group box B (HMGB) proteins are pivotal in the development of cancer. Although the proteomics of prostate cancer (PCa) cells has been reported, the involvement of HMGB proteins and their interactome in PCa is an unexplored field of considerable interest. We describe herein the results of the first HMGB1/HMGB2 interactome approach to PCa. Libraries constructed from the PCa cell line, PC-3, and from patients’ PCa primary tumor have been screened by the yeast 2-hybrid approach (Y2H) using HMGB1 and HMGB2 baits. Functional significance of this PCa HMGB interactome has been validated through expression and prognosis data available on public databases. Copy number alterations (CNA) affecting these newly described HMGB interactome components are more frequent in the most aggressive forms of PCa: those of neuroendocrine origin or castration-resistant PCa. Concordantly, adenocarcinoma PCa samples showing CNA in these genes are also associated with the worse prognosis. These findings open the way to their potential use as discriminatory biomarkers between high and low risk patients. Gene expression of a selected set of these interactome components has been analyzed by qPCR after HMGB1 and HMGB2 silencing. The data show that HMGB1 and HMGB2 control the expression of several of their interactome partners, which might contribute to the orchestrated action of these proteins in PCa.