RESUMEN
Suicide is defined as the action of harming oneself with the intention of dying. It is estimated that worldwide, one person dies by suicide every 40 s, making it a major health problem. Studies in families have suggested that suicide has a genetic component, so the search for genetic variants associated with suicidal behavior could be useful as potential biomarkers to identify people at risk of suicide. In Mexico, some studies of gene variants related to neurotransmission and other important pathways have been carried out and potential association of variants located in the following genes has been suggested: SLC6A4, SAT-1, TPH-2, ANKK1, GSHR, SCARA50, RGS10, STK33, COMT, and FKBP5. This systematic review shows the genetic studies conducted on the Mexican population. This article contributes by compiling the existing information on genetic variants and genes associated with suicidal behavior, in the future could be used as potential biomarkers to identify people at risk of suicide.
Asunto(s)
Proteínas RGS , Suicidio , Humanos , México/epidemiología , Ideación Suicida , Biomarcadores , Proteínas de Transporte de Serotonina en la Membrana Plasmática , Proteínas Serina-Treonina QuinasasRESUMEN
Plant survival depends on adaptive mechanisms that constantly rely on signal recognition and transduction. The predominant class of signal discriminators is receptor kinases, with a vast member composition in plants. The transduction of signals occurs in part by a simple repertoire of heterotrimeric G proteins, with a core composed of α-, ß-, and γ-subunits, together with a 7-transmembrane Regulator G Signaling (RGS) protein. With a small repertoire of G proteins in plants, phosphorylation by receptor kinases is critical in regulating the active state of the G-protein complex. This review describes the in vivo detected phosphosites in plant G proteins and conservation scores, and their in vitro corresponding kinases. Furthermore, recently described outcomes, including novel arrestin-like internalization of RGS and a non-canonical phosphorylation switching mechanism that drives G-protein plasticity, are discussed.
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Proteínas de Arabidopsis , Arabidopsis , Proteínas de Unión al GTP Heterotriméricas , Proteínas RGS , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Subunidades alfa de la Proteína de Unión al GTP/metabolismo , Proteínas de Unión al GTP Heterotriméricas/metabolismo , Fosforilación , Fosfotransferasas/metabolismo , Proteínas de Plantas/metabolismo , Plantas/metabolismo , Proteínas RGS/genética , Proteínas RGS/metabolismoRESUMEN
G protein-coupled receptors kinase 2 (GRK2) plays a major role in receptor regulation and, as a consequence, in cell biology and physiology. GRK2-mediated receptor desensitization is performed by its kinase domain, which exerts receptor phosphorylation promoting G protein uncoupling and the cessation of signaling, and by its RGS homology (RH) domain, able to interrupt G protein signaling. Since GRK2 activity is exacerbated in several pathologies, many efforts to develop inhibitors have been conducted. Most of them were directed toward GRK2 kinase activity and showed encouraging results on in vitro systems and animal models. Nevertheless, limitations including unspecific effects or pharmacokinetics issues prevented them from advancing to clinical trials. Surprisingly, even though the RH domain demonstrated the ability to desensitize GPCRs, this domain has been less explored. Herein, we show in vitro activity of a series of compounds that, by inhibiting GRK2 RH domain, increase receptor cAMP response, avoid GRK2 translocation to the plasma membrane, inhibit coimmunoprecipitation of GRK2 with Gαs subunit of heterotrimeric G protein, and prevent receptor desensitization. Also, we preliminarily evaluated candidates' ADMET properties and observed suitable lipophilicity and cytotoxicity. These novel inhibitors of phosphorylation-independent actions of GRK2 might be useful in elucidating other RH domain roles and lay the foundation for the development of innovative pharmacologic therapy for diseases where GRK2 activity is exacerbated.
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AMP Cíclico/metabolismo , Quinasa 2 del Receptor Acoplado a Proteína-G/antagonistas & inhibidores , Línea Celular Tumoral , Desarrollo de Medicamentos , Quinasa 2 del Receptor Acoplado a Proteína-G/metabolismo , Células HEK293 , Humanos , Fosforilación , Dominios Proteicos/efectos de los fármacos , Proteínas RGS/metabolismo , Transducción de Señal/efectos de los fármacosRESUMEN
The aim of this study was to characterize the morphological properties of amorphous silica nanoparticles (SiO2 NPs), their cytotoxicity and intracellular location within Human Osteoblasts (HOB). Additionally, SiO2 NPs were explored for their effectivity as carriers of CRTC3-siRNA on Human Preadipocytes (HPAd), and thus downregulate RGS2 gene expression. SiO2 NPs were synthesized using the method of Stöber at 45 °C, 56 °C, and 62 °C. These were characterized via TEM with EDS, Zeta Potential and FT-IR. Cytotoxicity was evaluated by XTT at three concentrations 50, 100 and 500 µg/mL; SiO2 NPs intracellular localization was observed through Confocal Laser Scanning Microscope. Delivering siRNA effectivity was measured by RT-qPCR. Morphology of SiO2 NPs was spherical with a range size from 64 to 119 nm; their surface charge was negative. Confocal images demonstrated that SiO2 NPs were located within cellular cytoplasm. At a SiO2 NPs concentration of 500 µg/mL HOB viability decreased, while at 50 µg/mL and 100 µg/mL cell viability was not affected regardless SiO2 NPs size. SiO2 NPs-CRTC3-siRNA are effective to down-regulate RGS2 gene expression in HPAd without cytotoxic effects. The developed SiO2 NPs-CRTC3-siRNA are a promising tool as a delivery vehicle to control obesity.
Asunto(s)
Nanopartículas/química , Proteínas RGS/metabolismo , ARN Interferente Pequeño/metabolismo , Dióxido de Silicio/química , Factores de Transcripción/farmacología , Antineoplásicos/farmacología , Supervivencia Celular/efectos de los fármacos , Regulación hacia Abajo , Sistemas de Liberación de Medicamentos , Técnicas de Silenciamiento del Gen , Humanos , Microscopía Confocal , Osteoblastos , Tamaño de la Partícula , Proteínas RGS/genética , Espectroscopía Infrarroja por Transformada de Fourier , Factores de Transcripción/genéticaRESUMEN
G protein-coupled receptors are critical mediators of platelet activation whose signaling can be modulated by members of the regulator of G protein signaling (RGS) family. The 2 most abundant RGS proteins in human and mouse platelets are RGS10 and RGS18. While each has been studied individually, critical questions remain about the overall impact of this mode of regulation in platelets. Here, we report that mice missing both proteins show reduced platelet survival and a 40% decrease in platelet count that can be partially reversed with aspirin and a P2Y12 antagonist. Their platelets have increased basal (TREM)-like transcript-1 expression, a leftward shift in the dose/response for a thrombin receptor-activating peptide, an increased maximum response to adenosine 5'-diphosphate and TxA2, and a greatly exaggerated response to penetrating injuries in vivo. Neither of the individual knockouts displays this constellation of findings. RGS10-/- platelets have an enhanced response to agonists in vitro, but platelet count and survival are normal. RGS18-/- mice have a 15% reduction in platelet count that is not affected by antiplatelet agents, nearly normal responses to platelet agonists, and normal platelet survival. Megakaryocyte number and ploidy are normal in all 3 mouse lines, but platelet recovery from severe acute thrombocytopenia is slower in RGS18-/- and RGS10-/-18-/- mice. Collectively, these results show that RGS10 and RGS18 have complementary roles in platelets. Removing both at the same time discloses the extent to which this regulatory mechanism normally controls platelet reactivity in vivo, modulates the hemostatic response to injury, promotes platelet production, and prolongs platelet survival.
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Plaquetas/metabolismo , Activación Plaquetaria/genética , Proteínas RGS/genética , Trombopoyesis/genética , Animales , Plaquetas/efectos de los fármacos , Supervivencia Celular/genética , Ratones , Ratones Noqueados , Fosforilación , Factor de Activación Plaquetaria/farmacología , Activación Plaquetaria/efectos de los fármacos , Inhibidores de Agregación Plaquetaria/farmacología , Recuento de Plaquetas , Proteínas RGS/metabolismo , Trombopoyesis/efectos de los fármacosRESUMEN
BACKGROUND: Evidence suggests that liability for suicide behavior is heritable; additionally, suicide has been partly related to other psychiatric disorders. Nevertheless, most of the information reported so far address Caucasian and Asian individuals. Hence, our aim was to conduct a gene-level association study in Mexican psychiatric individuals diagnosed with suicide attempt. METHODS: We recruited 192 individuals from two clinical centers in Mexico. All participants were born in Mexico and had Mexican parents and grandparents. Direct genotyping was performed using the commercial platform Infinium PsychArray BeadChip. A p-value lower than 1e-05 was considered as gene-level significant and a p-value lower than 1e-04 was considered as gene-level nominal significant. RESULTS: Our analyses showed that SCARA5 was associated to suicide intent at a gene-level with statistical significance (p-value = 1.12e-6). Other genes were nominally associated with suicide attempt: GHSR (p-value = 0.0004), RGS10 (p-value = 5.13e-5), and STK33 (p-value = 3.62e-5). Regarding gene variant analyses, the SNPs with a statistical association (p > .05) were rs561361616, rs1537577, rs11198999 for RGS10, and rs11041981, rs11041993, rs11041994, rs11041995, rs11041997, rs10840083, rs10769918 for STK33. For these genes, previous studies have associated SCARA5 with depression, GHSR with alcohol dependence and depression, and RGS10 with schizophrenia and depression. To date, STK33 has not been associated with any psychiatric disorder. CONCLUSION: Our outcomes revealed that SCARA5, GHSR, RGS10 and STK33 could be considered as risk biomarkers for suicide attempt behavior in our Mexican psychiatric sample. We recommend to perform larger scale analyses to have conclusive results.
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Estudio de Asociación del Genoma Completo/métodos , Polimorfismo de Nucleótido Simple , Intento de Suicidio/estadística & datos numéricos , Adulto , Predisposición Genética a la Enfermedad , Humanos , México/epidemiología , Persona de Mediana Edad , Proteínas Serina-Treonina Quinasas/genética , Proteínas RGS/genética , Receptores de Ghrelina/genética , Receptores Depuradores de Clase A/genética , Intento de Suicidio/psicología , Adulto JovenRESUMEN
Saccharomyces cerevisiae mitoribosomes are specialized in the translation of a few number of highly hydrophobic membrane proteins, components of the oxidative phosphorylation system. Mitochondrial characteristics, such as the membrane system and its redox state driven mitoribosomes evolution through great diversion from their bacterial and cytosolic counterparts. Therefore, mitoribosome presents a considerable number of mitochondrial-specific proteins, as well as new protein extensions. In this work we characterize temperature sensitive mutants of the subunit bL34 present in the 54S large subunit. Although bL34 has bacterial homologs, in yeast it has a long 65 aminoacids mitochondrial N-terminal addressing sequence, here we demonstrate that it can be replaced by the mitochondrial addressing sequence of Neurospora crassa ATP9 gene. The bL34 temperature sensitive mutants present lowered translation of mitochondrial COX1 and COX3, which resulted in reduced cytochrome c oxidase activity and respiratory growth deficiency. The sedimentation properties of bL34 in sucrose gradients suggest that similarly to its bacterial homolog, bL34 is also a later participant in the process of mitoribosome biogenesis.
Asunto(s)
Complejo IV de Transporte de Electrones/metabolismo , Mitocondrias/metabolismo , Ribosomas Mitocondriales/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Secuencia de Aminoácidos , Complejo IV de Transporte de Electrones/genética , Mitocondrias/genética , Proteínas Mitocondriales/genética , Proteínas Mitocondriales/metabolismo , Mutagénesis Sitio-Dirigida , Biosíntesis de Proteínas , Proteínas RGS/genética , Proteínas RGS/metabolismo , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/genética , Alineación de SecuenciaRESUMEN
BACKGROUND: Understanding the molecular basis underlying the formation of bone-forming osteocytes and lipid-storing adipocytes will help provide insights into the cause of disorders originating in stem/progenitor cells and develop therapeutic treatments for bone- or adipose-related diseases. In this study, the role of RGS2 and RGS4, two members of the regulators of G protein signaling (RGS) family, was investigated during adipogenenic and osteogenenic differentiation of human mesenchymal stem cells (hMSCs). RESULTS: Expression of RGS2 and RGS4 were found to be inversely regulated during adipogenesis induced by dexamethasone (DEX) and 3-isobutyl-methylxanthine, regardless if insulin was present, with RGS2 up-regulated and RGS4 down-regulated in response to adipogenic induction. RGS2 expression was also up-regulated during osteogenesis at a level similar to that induced by treatment of DEX alone, a shared component of adipogenic and osteogenic differentiation inducing media, but significantly lower than the level induced by adipogenic inducing media. RGS4 expression was down-regulated during the first 48 h of osteogenesis but up-regulated afterwards, in both cases at levels similar to that induced by DEX alone. Expression knock-down using small interfering RNA against RGS2 resulted in decreased differentiation efficiency during both adipogenesis and osteogenesis. On the other hand, expression knock-down of RGS4 also resulted in decreased adipogenic differentiation but increased osteogenic differentiation. CONCLUSIONS: RGS2 and RGS4 are differentially regulated during adipogenic and osteogenic differentiation of hMSCs. In addition, both RGS2 and RGS4 play positive roles during adipogenesis but opposing roles during osteogenesis, with RGS2 as a positive regulator and RGS4 as a negative regulator. These results imply that members of RGS proteins may play multifaceted roles during human adipogenesis and osteogenesis to balance or counterbalance each other's function during those processes.
Asunto(s)
Adipogénesis/fisiología , Regulación de la Expresión Génica/fisiología , Células Madre Mesenquimatosas/citología , Osteocitos/citología , Osteogénesis/fisiología , Proteínas RGS/metabolismo , Adipogénesis/genética , Regulación de la Expresión Génica/genética , Humanos , Osteogénesis/genética , Proteínas RGS/genética , Factores de TiempoRESUMEN
BACKGROUND: Understanding the molecular basis underlying the formation of bone-forming osteocytes and lipid-storing adipocytes will help provide insights into the cause of disorders originating in stem/progenitor cells and develop therapeutic treatments for bone- or adipose-related diseases. In this study, the role of RGS2 and RGS4, two members of the regulators of G protein signaling (RGS) family, was investigated during adipogenenic and osteogenenic differentiation of human mesenchymal stem cells (hMSCs). RESULTS: Expression of RGS2 and RGS4 were found to be inversely regulated during adipogenesis induced by dexamethasone (DEX) and 3-isobutyl-methylxanthine, regardless if insulin was present, with RGS2 up-regulated and RGS4 down-regulated in response to adipogenic induction. RGS2 expression was also up-regulated during osteogenesis at a level similar to that induced by treatment of DEX alone, a shared component of adipogenic and osteogenic differentiation inducing media, but significantly lower than the level induced by adipogenic inducing media. RGS4 expression was down-regulated during the first 48 h of osteogenesis but up-regulated afterwards, in both cases at levels similar to that induced by DEX alone. Expression knock-down using small interfering RNA against RGS2 resulted in decreased differentiation efficiency during both adipogenesis and osteogenesis. On the other hand, expression knock-down of RGS4 also resulted in decreased adipogenic differentiation but increased osteogenic differentiation. CONCLUSIONS: RGS2 and RGS4 are differentially regulated during adipogenic and osteogenic differentiation of hMSCs. In addition, both RGS2 and RGS4 play positive roles during adipogenesis but opposing roles during osteogenesis, with RGS2 as a positive regulator and RGS4 as a negative regulator. These results imply that members of RGS proteins may play multifaceted roles during human adipogenesis and osteogenesis to balance or counterbalance each other's function during those processes.
Asunto(s)
Humanos , Osteocitos/citología , Osteogénesis/fisiología , Regulación de la Expresión Génica/fisiología , Proteínas RGS/metabolismo , Adipogénesis/fisiología , Células Madre Mesenquimatosas/citología , Osteogénesis/genética , Factores de Tiempo , Regulación de la Expresión Génica/genética , Proteínas RGS/genética , Adipogénesis/genéticaRESUMEN
According to receptor theory, the effect of a ligand depends on the amount of agonist-receptor complex. Therefore, changes in receptor abundance should have quantitative effects. However, the response to pheromone in Saccharomyces cerevisiae is robust (unaltered) to increases or reductions in the abundance of the G-protein-coupled receptor (GPCR), Ste2, responding instead to the fraction of occupied receptor. We found experimentally that this robustness originates during G-protein activation. We developed a complete mathematical model of this step, which suggested the ability to compute fractional occupancy depends on the physical interaction between the inhibitory regulator of G-protein signaling (RGS), Sst2, and the receptor. Accordingly, replacing Sst2 by the heterologous hsRGS4, incapable of interacting with the receptor, abolished robustness. Conversely, forcing hsRGS4:Ste2 interaction restored robustness. Taken together with other results of our work, we conclude that this GPCR pathway computes fractional occupancy because ligand-bound GPCR-RGS complexes stimulate signaling while unoccupied complexes actively inhibit it. In eukaryotes, many RGSs bind to specific GPCRs, suggesting these complexes with opposing activities also detect fraction occupancy by a ratiometric measurement. Such complexes operate as push-pull devices, which we have recently described.
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Proteínas Activadoras de GTPasa/metabolismo , Receptores del Factor de Conjugación/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/metabolismo , Humanos , Modelos Teóricos , Unión Proteica , Proteínas RGS/metabolismoRESUMEN
To identify single-nucleotide polymorphisms that contribute to the genetic susceptibility to schizophrenia, we examined the potential association between schizophrenia and 9 single nucleotide polymorphisms (rs1530351, rs4791230, rs2869577, rs8077696, rs8070231, rs2292592, rs9916525, rs1122079, and rs4790953) in the G-protein signaling 9 gene. The participants included 395 schizophrenia subjects and 400 healthy controls. The selected single nucleotide polymorphisms were genotyped using mass spectrometry techniques. The allelic or genotypic frequencies of the rs4791230 (promoter region) polymorphisms in subjects with schizophrenia were significantly different from those in healthy controls. The subjects with schizophrenia had a significantly higher frequency of the G allele (P = 0.030, odds ratio = 1.589, 95% confidence interval = 1.042-2.422) of rs4791230. Strong linkage disequilibrium was observed in 4 blocks (D' > 0.9). Significantly fewer T-A (rs1530351-rs4791230) haplotypes (P = 0.029) were found in subjects with schizophrenia. These findings suggest a role of G-protein signaling 9 polymorphisms in schizophrenia among Han Chinese and may be informative for future genetic or neurobiological studies on schizophrenia.
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Proteínas RGS/genética , Esquizofrenia/genética , Adulto , Pueblo Asiatico/genética , Estudios de Casos y Controles , China , Femenino , Proteínas de Unión al GTP/genética , Estudios de Asociación Genética , Predisposición Genética a la Enfermedad , Variación Genética , Haplotipos , Humanos , Desequilibrio de Ligamiento , Masculino , Persona de Mediana Edad , Polimorfismo de Nucleótido Simple , Proteínas RGS/metabolismo , Esquizofrenia/metabolismoRESUMEN
Throughout evolution, organisms have created numerous mechanisms to sense and respond to their environment. One such highly conserved mechanism involves regulation by heterotrimeric G-protein complex comprised of alpha (Gα), beta (Gß) and gamma (Gγ) subunits. In plants, these proteins play important roles in signal transduction pathways related to growth and development including response to biotic and abiotic stresses and consequently affect yield. In this work, we have identified and characterized the complete heterotrimeric G-protein repertoire in the Capsicum annuum (Capsicum) genome which consists of one Gα, one Gß and three Gγ genes. We have also identified one RGS gene in the Capsicum genome that acts as a regulator of the G-protein signaling. Biochemical activities of the proteins were confirmed by assessing the GTP-binding and GTPase activity of the recombinant Gα protein and its regulation by the GTPase acceleration activity of the RGS protein. Interaction between different subunits was established using yeast- and plant-based analyses. Gene and protein expression profiles of specific G-protein components revealed interesting spatial and temporal regulation patterns, especially during root development and during fruit development and maturation. This research thus details the characterization of the first heterotrimeric G-protein family from a domesticated, commercially important vegetable crop.
Asunto(s)
Transducción de Señal , Solanaceae/genética , Capsicum/genética , Capsicum/crecimiento & desarrollo , Capsicum/metabolismo , Frutas/genética , Frutas/crecimiento & desarrollo , Frutas/metabolismo , Proteínas de Unión al GTP/genética , Proteínas de Unión al GTP/metabolismo , Regulación del Desarrollo de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Proteínas de Unión al GTP Heterotriméricas/genética , Proteínas de Unión al GTP Heterotriméricas/metabolismo , Especificidad de Órganos , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Mapeo de Interacción de Proteínas , Proteínas RGS/genética , Proteínas RGS/metabolismoRESUMEN
The entomopathogenic fungus Beauveria bassiana is widely used in pest biocontrol strategies. We evaluated both the antioxidant response mediated by compatible solutes, trehalose or mannitol, and the expression of related genes using oxygen pulses at three oxygen concentrations in solid state culture (SSC): normal atmosphere (21% O2), low oxygen (16% O2) and enriched oxygen (26% O2). Trehalose concentration decreased 75% after atmospheric modifications in the cultures, whereas mannitol synthesis was three-fold higher under the 16% O2 pulses relative to normal atmosphere (100 and 30 µg mannitol mg(-1) biomass, respectively). Confirming this result, expression of the mpd gene, coding for mannitol-1-P dehydrogenase (MPD), increased up to 1.4 times after O2 pulses. The expression of the bbrgs1 gene, encoding a regulatory G protein related to conidiation, was analysed to explain previously reported differences in conidial production. Surprisingly, expression of bbrgs1 decreased after atmospheric modification. Finally, principal component analysis (PCA) indicated that 83.39% of the variability in the data could be explained by two components. This analysis corroborated the positive correlation between mannitol concentration and mpd gene expression, as well as the negative correlation between conidial production and bbrgs1 gene expression. This study contributes to understanding of antioxidant and molecular response of B. bassiana induced under oxidant conditions.
Asunto(s)
Antioxidantes/metabolismo , Beauveria/efectos de los fármacos , Beauveria/metabolismo , Oxígeno/metabolismo , Estrés Fisiológico , Beauveria/genética , Beauveria/crecimiento & desarrollo , Medios de Cultivo/química , Perfilación de la Expresión Génica , Manitol/metabolismo , Proteínas RGS/biosíntesis , Proteínas RGS/genética , Esporas Fúngicas/crecimiento & desarrollo , Deshidrogenasas del Alcohol de Azúcar/biosíntesis , Deshidrogenasas del Alcohol de Azúcar/genética , Trehalosa/metabolismoRESUMEN
Genetic variants of the RGS5 gene are believed to be risk factors for hypertension and cardiovascular diseases. In this study, we investigated the association between RGS5 gene variants and hypertension in the Mongolian and Han populations. Peripheral blood was obtained from 429 unrelated Mongolian herdsmen and 416 Han farmers [including essential hypertension (EH) patients and controls]. Nine tagSNPs within the RGS5 genes were retrieved from HapMap, and the samples were individually genotyped using the polymerase chain reaction/ligase detection reaction assay. The distribution of the allele frequency of rs12035879 differed significantly between hypertensive subjects and controls in the Han population, while the distribution of the allele and genotype frequencies of rs16849802 differed significantly between hypertensive subjects and controls in the Mongolian population. We observed an association between rs16849802 and EH in the Mongolian population. The frequency of haplotype GAA was significantly higher in the EH group than in controls in the Mongolian population. However, the EH group and controls did not differ significantly in all 6 haplotypes in the Han population. The rs16849802 and haplotype GAA independently increased the risk of EH in Mongolian patients, and may be used as a risk factor for the prediction of high blood pressure.
Asunto(s)
Estudios de Asociación Genética , Hipertensión/genética , Polimorfismo de Nucleótido Simple/genética , Proteínas RGS/genética , Adulto , Anciano , China , Hipertensión Esencial , Femenino , Frecuencia de los Genes , Genotipo , Haplotipos , Humanos , Hipertensión/patología , Masculino , Persona de Mediana Edad , Mongolia , Factores de RiesgoRESUMEN
Multidrug resistance (MDR) poses a serious impediment to the success of chemotherapy for laryngeal cancer. To identify microRNAs and mRNAs associated with MDR of human laryngeal cancer Hep-2 cells, we developed a multidrug-resistant human laryngeal cancer subline, designated Hep-2/v, by exposing Hep-2 cells to stepwise increasing concentrations of vincristine (0.02-0.96'µM). Microarray assays were performed to compare the microRNA and mRNA expression profiles of Hep-2 and Hep-2/v cells. Compared to Hep-2 cells, Hep-2/v cells were more resistant to chemotherapy drugs (∼45-fold more resistant to vincristine, 5.1-fold more resistant to cisplatin, and 5.6-fold more resistant to 5-fluorouracil) and had a longer doubling time (42.33±1.76 vs 28.75±1.12'h, P<0.05), higher percentage of cells in G0/G1 phase (80.98±0.52 vs 69.14±0.89, P<0.05), increased efflux of rhodamine 123 (95.97±0.56 vs 12.40±0.44%, P<0.01), and up-regulated MDR1 expression. A total of 7 microRNAs and 605 mRNAs were differentially expressed between the two cell types. Of the differentially expressed mRNAs identified, regulator of G-protein signaling 10, high-temperature requirement protein A1, and nuclear protein 1 were found to be the putative targets of the differentially expressed microRNAs identified. These findings may open a new avenue for clarifying the mechanisms responsible for MDR in laryngeal cancer.
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Humanos , Resistencia a Antineoplásicos/genética , Neoplasias Laríngeas/genética , MicroARNs/aislamiento & purificación , Miembro 1 de la Subfamilia B de Casetes de Unión a ATP/genética , ARN Mensajero/aislamiento & purificación , Antineoplásicos/farmacología , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/genética , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Cisplatino/farmacología , Citometría de Flujo , Fluorouracilo/farmacología , Puntos de Control de la Fase G1 del Ciclo Celular , Genes MDR , Neoplasias Laríngeas/tratamiento farmacológico , Proteínas de Neoplasias/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Proteínas RGS/genética , /farmacocinética , Serina Endopeptidasas/genética , Análisis de Matrices Tisulares , Vincristina/farmacologíaRESUMEN
Multidrug resistance (MDR) poses a serious impediment to the success of chemotherapy for laryngeal cancer. To identify microRNAs and mRNAs associated with MDR of human laryngeal cancer Hep-2 cells, we developed a multidrug-resistant human laryngeal cancer subline, designated Hep-2/v, by exposing Hep-2 cells to stepwise increasing concentrations of vincristine (0.02-0.96'µM). Microarray assays were performed to compare the microRNA and mRNA expression profiles of Hep-2 and Hep-2/v cells. Compared to Hep-2 cells, Hep-2/v cells were more resistant to chemotherapy drugs (≈ 45-fold more resistant to vincristine, 5.1-fold more resistant to cisplatin, and 5.6-fold more resistant to 5-fluorouracil) and had a longer doubling time (42.33 ± 1.76 vs 28.75 ± 1.12'h, P<0.05), higher percentage of cells in G0/G1 phase (80.98 ± 0.52 vs 69.14 ± 0.89, P<0.05), increased efflux of rhodamine 123 (95.97 ± 0.56 vs 12.40 ± 0.44%, P<0.01), and up-regulated MDR1 expression. A total of 7 microRNAs and 605 mRNAs were differentially expressed between the two cell types. Of the differentially expressed mRNAs identified, regulator of G-protein signaling 10, high-temperature requirement protein A1, and nuclear protein 1 were found to be the putative targets of the differentially expressed microRNAs identified. These findings may open a new avenue for clarifying the mechanisms responsible for MDR in laryngeal cancer.
Asunto(s)
Miembro 1 de la Subfamilia B de Casetes de Unión a ATP/genética , Resistencia a Antineoplásicos/genética , Neoplasias Laríngeas/genética , MicroARNs/aislamiento & purificación , ARN Mensajero/aislamiento & purificación , Subfamilia B de Transportador de Casetes de Unión a ATP , Antineoplásicos/farmacología , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/genética , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Cisplatino/farmacología , Citometría de Flujo , Fluorouracilo/farmacología , Puntos de Control de la Fase G1 del Ciclo Celular , Genes MDR , Serina Peptidasa A1 que Requiere Temperaturas Altas , Humanos , Neoplasias Laríngeas/tratamiento farmacológico , Proteínas de Neoplasias/genética , Proteínas RGS/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Rodamina 123/farmacocinética , Serina Endopeptidasas/genética , Análisis de Matrices Tisulares , Vincristina/farmacologíaRESUMEN
Recent studies demonstrate an association between antipsychotic-induced parkinsonism (AIP) and rs4606 SNP of RGS2 gene in Jewish and African-Americans. The current study investigates the association between rs4606 and AIP or its subsymptoms (rest tremor, rigidity, and bradykinesia) in 112 psychiatric inpatients of African-Caribbean origin. Presence of AIP, rigidity, bradykinesia, and tremor was measured by the UPDRS. We applied chi(2) (or Fisher Exact) and logistic regression analyses in several models including rs4606, age, gender, dose of antipsychotics, and anticholinergics, and two other putatively functional SNPs in DRD2 (-141CIns/Del) and HTR2C (Cys23Ser) genes. In contrast to recent literature, we find no evidence for an association between rs4606 and AIP or any of its subsymptoms. We hypothesize that the observed lack of association is due probably to differences in serotonin 2A-receptor affinities of the antipsychotics utilized (in contrast to the other published studies, the majority of our patients utilized typical antipsychotics).
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Antipsicóticos/efectos adversos , Trastornos Parkinsonianos/genética , Polimorfismo de Nucleótido Simple , Proteínas RGS/genética , Adulto , Factores de Edad , Antipsicóticos/administración & dosificación , Antipsicóticos/farmacología , Población Negra , Relación Dosis-Respuesta a Droga , Femenino , Humanos , Modelos Logísticos , Masculino , Persona de Mediana Edad , Trastornos Parkinsonianos/inducido químicamente , Escalas de Valoración Psiquiátrica , Receptor de Serotonina 5-HT2A/metabolismo , Factores de Riesgo , Factores Sexuales , Indias OccidentalesRESUMEN
Linkage and association studies in five independently ascertained samples have suggested that polymorphisms of the regulator of G-protein signaling 4 (RGS4) may confer risk for schizophrenia (SCZ). Suggestive evidence for association with bipolar disorder (BD) has also been presented. However, the associated alleles and haplotypes have differed among the samples. Data from other independent samples may clarify the putative associations. Hence, we investigated an independent, ethnically diverse Brazilian population comprising patients with SCZ (n=271) or BD1 (n=306), who were contrasted with 576 community-based controls. Parents of 49 SCZ cases and 44 BD cases were available for transmission disequilibrium tests (TDTs). Four RGS4 single-nucleotide polymorphisms (SNPs) 1, 4, 7 and 18 putatively associated with SCZ were investigated. In the SCZ samples, significant case-control differences were not observed for individual SNPs or haplotypes, though the TDT suggested transmission distortion similar to that observed in the initial report. For the BD sample, case-control comparisons revealed no significant differences for individual SNPs, but an omnibus test suggested differences in the overall distribution of haplotypes bearing all four SNPs (SNP-EM Omnibus likelihood ratio test; P=0.003). The TDT revealed over-transmission of allele A at SNP7 (P=0.016), as well as haplotypes incorporating this allele. However, global tests incorporating all haplotypes yielded only suggestive trends for association (P=0.19). In conclusion, association with SCZ was not detected in the present analyses. The failure to detect an association may be related to inadequate power or to confounds related to ethnic admixture. Suggestive associations with BD detected here require further investigation in a larger sample.
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Trastorno Bipolar/genética , Haplotipos , Polimorfismo de Nucleótido Simple/genética , Proteínas RGS/genética , Esquizofrenia/genética , Adulto , Brasil , Estudios de Casos y Controles , Femenino , Predisposición Genética a la Enfermedad , Humanos , Desequilibrio de Ligamiento , Masculino , Persona de Mediana Edad , Linaje , Valores de ReferenciaRESUMEN
Follicle-stimulating hormone receptor (FSHR) and luteinizing hormone receptor (LHR) belong to the super-family of G protein-coupled receptors (GPCR); GPCRs are negatively regulated by RGS ("regulators of G protein signaling") proteins. In this study we evaluated the effects of RGS3 and RGS10 on FSHR and LHR ligand binding and effector coupling. FSHR and LHR ligand binding were unchanged in the presence of RGS3 or RGS10. However, signaling by FSHR and LHR was diminished by RGS3 but not by RGS10. This constitutes the first demonstration of an interaction between RGS proteins and LH and FSH signaling pathways and identifies a mechanism for negative regulation of RGS3 on FSHR and LHR signaling.
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Proteínas RGS/farmacología , Receptores de HFE/metabolismo , Receptores de HL/metabolismo , Proteínas Represoras/farmacología , Transducción de Señal/fisiología , Animales , Línea Celular , Femenino , Humanos , Ligandos , Ratas , Ratas Sprague-Dawley , Transducción de Señal/efectos de los fármacosRESUMEN
We applied a systematic bioinformatics approach, followed by careful manual inspection and experimental validation to identify additional expressed sequences located at the Hereditary Prostate Cancer Region (HPC1) between D1S2818 and D1S1642 on chromosome 1q25. All transcripts already described for the 1q25 region were identified and we were able to define 11 additional expressed sequences within this region (three full-length cDNA clone sequences and eight ESTs), increasing the total number of gene count in this region by 38%. Five out of the 11 expressed sequences identified were shown to be expressed in prostate tissue and thus represent novel disease gene candidates for the HPC1 region. Here, we report a detailed characterization of these five novel disease gene candidates, their expression pattern in various tissues, their genomic organization and functional annotation. Two candidates (RGSL1 and RGSL2) correspond to novel members of the RGS family, which is involved in the regulation of G-protein signaling. RGSL1 and RGLS2 expression was detected by real-time polymerase chain reaction in normal prostate tissue, but could not be detected in prostate tumor cell lines, suggesting they might have a role in prostate cancer.