RESUMO
This article is a report of the "International Colloquium on Gap junctions: 50Years of Impact on Cancer" that was held 8-9 September 2016, at the Amphitheater "Pôle Biologie Santé" of the University of Poitiers (Poitiers, France). The colloquium was organized by M Mesnil (Université de Poitiers, Poitiers, France) and C Naus (University of British Columbia, Vancouver, Canada) to celebrate the 50th anniversary of the seminal work published in 1966 by Loewenstein and Kanno [Intercellular communication and the control of tissue growth: lack of communication between cancer cells, Nature, 116 (1966) 1248-1249] which initiated studies on the involvement of gap junctions in carcinogenesis. During the colloquium, 15 participants presented reviews or research updates in the field which are summarized below.
Assuntos
Junções Comunicantes/metabolismo , Proteínas de Neoplasias/metabolismo , Neoplasias/metabolismo , Animais , Junções Comunicantes/genética , Junções Comunicantes/patologia , Humanos , Proteínas de Neoplasias/genética , Neoplasias/genética , Neoplasias/patologiaRESUMO
OBJECTIVE: Using non-invasive magnetic resonance (MR) techniques and a histological approach, we assessed the outcomes of perinatal exposure at a low dose of 3,3'-DCBPA (2-chloro-4-[1-(3-chloro-4-hydroxyphenyl)-1-methylethyl]phenol) and/or 3,5-DCBPA (2,6-dichloro-4-[1-(4-hydroxyphenyl)-1-methylethyl]phenol) on mice livers. MATERIALS AND METHODS: Fertilized female Swiss mice were injected intraperitoneally during gestation and lactation with either vehicle control, 20 µg/kg/day of BPA, 3,5-DCBPA, 3,3'-DCBPA or a mixture (mix-DCBPA). Complementary methods were used to evaluate, in male and female pups, (1) liver structure by texture analysis of images obtained through MR imaging (MRI) and histology, (2) hepatic lipid composition through in vivo 1H MR spectroscopy (1H MRS). RESULTS: Principal component analysis of texture parameters showed no structural modification of the liver with BPA and DCBPA treatments. Accordingly, no hepatic microvesicular steatosis was observed through hematoxylin-eosin staining. Compared to control, MRS revealed no difference in lipid composition for BPA, 3,5-DCBPA or 3,3'-DCBPA groups. However, MRS detected a significant increase in the mix-DCBPA groups for the saturated component of fatty acids (FA), total unsaturated FA bond index and polyunsaturated FA bond index. CONCLUSION: Prior to any structural changes, polyunsaturated fatty acids significantly increased in young male and female mice exposed perinatally at a low dose to a mixture of dichlorinated BPA.
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Compostos Benzidrílicos/toxicidade , Lipídeos/análise , Fígado/efeitos dos fármacos , Espectroscopia de Ressonância Magnética , Exposição Materna , Fenóis/toxicidade , Animais , Peso Corporal , Ácidos Graxos , Fígado Gorduroso , Feminino , Lactação , Fígado/diagnóstico por imagem , Fígado/patologia , Imageamento por Ressonância Magnética , Masculino , Camundongos , GravidezRESUMO
Lipid rafts are specific microdomains of plasma membrane which are enriched in cholesterol and sphingolipids. These domains seem to favour the interactions of particular proteins and the regulation of signalling pathways in the cells. Recent data have shown that among the proteins, which are preferentially localized in lipid rafts, are connexins that are the structural proteins of gap junctions. Since gap junctional intercellular communication is involved in various cellular processes and pathologies such as cancer, we were interested to review the various observations concerning this specific localization of connexins in lipid rafts and its consequences on gap junctional intercellular communication capacity. In particular, we will focus our discussion on the role of the lipid raft-connexin connection in cancer progression. This article is part of a Special Issue entitled: The Communicating junctions, composition, structure and characteristics.
Assuntos
Junções Comunicantes/fisiologia , Microdomínios da Membrana/química , Animais , Comunicação Celular , Movimento Celular , Conexinas/química , Junções Comunicantes/metabolismo , Humanos , Lipídeos/química , Microdomínios da Membrana/fisiologia , Modelos Biológicos , Modelos Moleculares , Neoplasias/metabolismo , Conformação Proteica , Transdução de SinaisRESUMO
Glioblastoma cells are characterized by high proliferation and invasive capacities. Tumor development has been associated with a decrease of gap-junctional intercellular communication, but the concrete involvement of gap junction proteins, connexins, remains elusive since they are also suspected to promote cell invasion. In order to better understand how connexins control the glioma cell phenotype, we studied the consequences of inhibiting the intrinsic expression of the major astrocytic connexin, Connexin43, in human U251 glioblastoma cells by the shRNA strategy. The induced down-regulation of Cx43 expression has various effects on the U251 cells such as increased clonogenicity, angiogenesis and decreased adhesion on specific extracellular matrix proteins. We demonstrate that the invasion capacity measured in vitro and ex vivo correlates with Cx43 expression level. For the first time in a cancer cell context, our work demonstrates that Cx43 cofractionates, colocalizes and coimmunoprecipitates with a lipid raft marker, caveolin-1 and that this interaction is inversely correlated to the level of Cx43. This localization of Cx43 in these lipid raft microdomains regulates both homo- and heterocellular gap junctional communications (respectively between U251 cells, or between U251 cells and astrocytes). Moreover, the adhesive and invasive capacities are not dependent, in our model, on Cav-1 expression level. Our results tend to show that heterocellular gap junctional communication between cancer and stroma cells may affect the behavior of the tumor cells. Altogether, our data demonstrate that Cx43 controls the tumor phenotype of glioblastoma U251 cells and in particular, invasion capacity, through its localization in lipid rafts containing Cav-1.
Assuntos
Caveolina 1/metabolismo , Conexina 43/genética , Conexina 43/metabolismo , Regulação para Baixo , Glioblastoma/genética , Invasividade Neoplásica/genética , Animais , Caveolina 1/análise , Adesão Celular , Comunicação Celular , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Células Cultivadas , Galinhas , Conexina 43/análise , Junções Comunicantes/genética , Junções Comunicantes/metabolismo , Junções Comunicantes/patologia , Regulação Neoplásica da Expressão Gênica , Glioblastoma/metabolismo , Glioblastoma/patologia , Humanos , Microdomínios da Membrana/genética , Microdomínios da Membrana/metabolismo , Microdomínios da Membrana/patologia , Camundongos , Invasividade Neoplásica/patologiaRESUMO
Serotonin (5-hydroxytryptamine, 5-HT) is a biogenic monoamine acting as a neurotransmitter in the central nervous system (CNS), local mediator in the gut, and vasoactive agent in the blood. It has been linked to a variety of CNS functions and is implicated in many CNS and psychiatric disorders. The high comorbidity between some neuropathies can be partially understood by the fact that these diseases share a common etiology involving the serotoninergic system. In addition to its well-known functions, serotonin has been shown to be a mitogenic factor for a wide range of normal and tumor cells, including glioma cells, in vitro. The developing CNS of fetus and newborn is particularly susceptible to the deleterious effects of neurotoxic substances in our environment, and perinatal exposure could result in the later development of diseases, a hypothesis known as the developmental origin of health and disease. Some of these substances affect the serotoninergic system and could therefore be the source of a silent pandemic of neurodevelopmental toxicity. This review presents the available data that are contributing to the appreciation of the effects of the exposome on the serotoninergic system and their potential link with brain pathologies (neurodevelopmental, neurodegenerative, neurobehavioral disorders, and glioblastoma).
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5-hydroxytryptamine-4 (5-HT(4)) receptors have been proposed to contribute to the generation of atrial fibrillation in human atrial myocytes, but it is unclear if these receptors are present in the hearts of small laboratory animals (e.g. rat). In this study, we examined presence and functionality of 5-HT(4) receptors in auricular myocytes of newborn rats and their possible involvement in regulation of gap junctional intercellular communication (GJIC, responsible for the cell-to-cell propagation of the cardiac excitation). Western-blotting assays showed that 5-HT(4) receptors were present and real-time RT-PCR analysis revealed that 5-HT(4b) was the predominant isoform. Serotonin (1 microM) significantly reduced cAMP concentration unless a selective 5-HT(4) inhibitor (GR113808 or ML10375, both 1 microM) was present. Serotonin also reduced the amplitude of L-type calcium currents and influenced the strength of GJIC without modifying the phosphorylation profiles of the different channel-forming proteins or connexins (Cxs), namely Cx40, Cx43 and Cx45. GJIC was markedly increased when serotonin exposure occurred in presence of a 5-HT(4) inhibitor but strongly reduced when 5-HT(2A) and 5-HT(2B) receptors were inhibited, showing that activation of these receptors antagonistically regulated GJIC. The serotoninergic response was completely abolished when 5-HT(4), 5-HT(2A) and 5-HT(2B) were simultaneously inhibited. A 24 h serotonin exposure strongly reduced Cx40 expression whereas Cx45 was less affected and Cx43 still less. In conclusion, this study revealed that 5-HT(4) (mainly 5-HT(4b)), 5-HT(2A) and 5-HT(2B) receptors coexisted in auricular myocytes of newborn rat, that 5-HT(4) activation reduced cAMP concentration, I(Ca)(L) and intercellular coupling whereas 5-HT(2A) or 5-HT(2B) activation conversely enhanced GJIC.
Assuntos
Junções Comunicantes/metabolismo , Átrios do Coração/citologia , Miócitos Cardíacos/metabolismo , Receptor 5-HT2A de Serotonina/metabolismo , Receptor 5-HT2B de Serotonina/metabolismo , Receptor 5-HT2C de Serotonina/metabolismo , Receptores 5-HT4 de Serotonina/metabolismo , Adenilil Ciclases/metabolismo , Aminobenzoatos/farmacologia , Animais , Animais Recém-Nascidos , Western Blotting , Células Cultivadas , Conexinas/metabolismo , Junções Comunicantes/efeitos dos fármacos , Técnicas In Vitro , Indóis/farmacologia , Miócitos Cardíacos/efeitos dos fármacos , Técnicas de Patch-Clamp , Fosforilação/efeitos dos fármacos , Piperidinas/farmacologia , Ratos , Ratos Wistar , Receptor 5-HT2A de Serotonina/genética , Receptor 5-HT2B de Serotonina/genética , Receptor 5-HT2C de Serotonina/genética , Receptores 5-HT4 de Serotonina/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Serotonina/farmacologia , Antagonistas do Receptor 5-HT2 de Serotonina , Antagonistas do Receptor 5-HT4 de Serotonina , Serotoninérgicos/farmacologia , Antagonistas da Serotonina/farmacologia , Sulfonamidas/farmacologia , para-AminobenzoatosRESUMO
Bone is a dynamic tissue that undergoes a precise remodeling process involving resorptive osteoclastic cells and bone-forming osteoblastic (OB) cells. The functional imbalance of either of these cell types can lead to severe skeletal diseases. The proliferation and differentiation of OB cells play a major role in bone development and turnover. These cellular processes are coordinated by connexin43 (Cx43)-based gap-junctional intercellular communication (GJIC) and by soluble factors such as endothelin-1 (ET-1). We have used the Cx43 heterozygous (Cx43(+/-)) murine model to study the possible cross-talk between Cx43 and ET-1 in cultured calvarial OB cells. On microcomputed tomographic analysis of 3-day-old pups, Cx43(+/-) mice showed hypomineralized calvaria in comparison with their Cx43(+/+) littermates. Characterization of cultured OB cells clearly demonstrated the effect of the partial deletion of the Cx43 gene on its expression, on GJIC, and subsequently on OB differentiation. In this model, ET-1 (10(-8) M) lost its mitogenic action in Cx43(+/-) OB cells compared with Cx43(+/+) cells. Moreover, a correlation between the inhibition of cell differentiation by ET-1 and the decreased amount and function of Cx43 was found in Cx43(+/+) OB cells but not in their Cx43(+/-) counterparts. Thus, as Cx43 is linked to OB differentiation, our data indicate that this mitogenic ET-1 peptide has pronounced effects on fully differentiated OB cells. With respect to roles in mechanotransduction and OB differentiation, Cx43 might modulate osteoblastic sensitivity to soluble factors.
Assuntos
Diferenciação Celular/genética , Conexina 43/metabolismo , Endotelina-1/metabolismo , Junções Comunicantes/metabolismo , Osteoblastos/metabolismo , Osteogênese/genética , Animais , Doenças Ósseas Metabólicas/genética , Remodelação Óssea/efeitos dos fármacos , Remodelação Óssea/genética , Calcificação Fisiológica/genética , Comunicação Celular/efeitos dos fármacos , Comunicação Celular/genética , Diferenciação Celular/efeitos dos fármacos , Divisão Celular/genética , Proliferação de Células , Conexina 43/genética , Endotelina-1/farmacologia , Junções Comunicantes/efeitos dos fármacos , Junções Comunicantes/genética , Inibidores do Crescimento/metabolismo , Inibidores do Crescimento/farmacologia , Mecanotransdução Celular/efeitos dos fármacos , Mecanotransdução Celular/fisiologia , Camundongos , Camundongos Knockout , Camundongos Transgênicos , Técnicas de Cultura de Órgãos , Osteoblastos/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Crânio/diagnóstico por imagem , Crânio/metabolismo , Crânio/fisiopatologia , Microtomografia por Raio-XRESUMO
The incidence of brain pathologies has increased during last decades. Better diagnosis (autism spectrum disorders) and longer life expectancy (Parkinson's disease, Alzheimer's disease) partly explain this increase, while emerging data suggest pollutant exposures as a possible but still underestimated cause of major brain disorders. Taking into account that the brain parenchyma is rich in gap junctions and that most pollutants inhibit their function; brain disorders might be the consequence of gap-junctional alterations due to long-term exposures to pollutants. In this article, this hypothesis is addressed through three complementary aspects: (1) the gap-junctional organization and connexin expression in brain parenchyma and their function; (2) the effect of major pollutants (pesticides, bisphenol A, phthalates, heavy metals, airborne particles, etc.) on gap-junctional and connexin functions; (3) a description of the major brain disorders categorized as neurodevelopmental (autism spectrum disorders, attention deficit hyperactivity disorders, epilepsy), neurobehavioral (migraines, major depressive disorders), neurodegenerative (Parkinson's and Alzheimer's diseases) and cancers (glioma), in which both connexin dysfunction and pollutant involvement have been described. Based on these different aspects, the possible involvement of pollutant-inhibited gap junctions in brain disorders is discussed for prenatal and postnatal exposures.
Assuntos
Doença de Alzheimer/genética , Encefalopatias/genética , Conexina 43/genética , Transtorno Depressivo Maior/metabolismo , Poluentes Atmosféricos/toxicidade , Doença de Alzheimer/induzido quimicamente , Doença de Alzheimer/patologia , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Encefalopatias/induzido quimicamente , Encefalopatias/patologia , Comunicação Celular/efeitos dos fármacos , Transtorno Depressivo Maior/induzido quimicamente , Transtorno Depressivo Maior/patologia , Feminino , Junções Comunicantes/efeitos dos fármacos , Junções Comunicantes/genética , Junções Comunicantes/patologia , Humanos , GravidezRESUMO
The resistance of glioblastomas to treatments is mainly the consequence of their invasive capacities. Therefore, in order to better treat these tumors, it is important to understand the molecular mechanisms which are responsible for this behavior. Previous work suggested that gap junction proteins, the connexins, facilitate the aggressive nature of glioma cells. Here, we show that one of them-connexin43 (Cx43)-is implicated in the formation and function of invadopodia responsible for invasion capacity of U251 human glioblastoma cells. Immunofluorescent approaches-combined with confocal analyses-revealed that Cx43 was detected in all the formation stages of invadopodia exhibiting proteolytic activity. Clearly, Cx43 appeared to be localized in invadopodia at low cell density and less associated with the establishment of gap junctions. Accordingly, lower extracellular matrix degradation correlated with less mature invadopodia and MMP2 activity when Cx43 expression was decreased by shRNA strategies. Moreover, the kinetics of invadopodia formation could be dependent on Cx43 dynamic interactions with partners including Src and cortactin. Interestingly, it also appeared that invadopodia formation and MMP2 activity are dependent on Cx43 hemichannel activity. In conclusion, these results reveal that Cx43 might be involved in the formation and function of the invadopodia of U251 glioblastoma cells.
Assuntos
Conexina 43/metabolismo , Glioblastoma/metabolismo , Glioblastoma/patologia , Podossomos/metabolismo , Linhagem Celular Tumoral , Gelatina/metabolismo , Técnicas de Silenciamento de Genes , Humanos , Modelos Biológicos , Ligação Proteica , ProteóliseRESUMO
Tissue homeostasis is the result of a complex intercellular network controlling the behavior of every cell for the survival of the whole organism. In mammalian tissues, cells do communicate via diverse long- and short-range communication mechanisms. While long-range communication involves hormones through blood circulation and neural transmission, short-range communication mechanisms include either paracrine diffusible factors or direct interactions (e.g., gap junctions, intercellular bridges and tunneling nanotubes) or a mixture of both (e.g., exosomes). Tumor growth represents an alteration of tissue homeostasis and could be the consequence of intercellular network disruption. In this network, direct short-range intercellular communication seems to be particularly involved. The first type of these intercellular communications thought to be involved in cancer progression were gap junctions and their protein subunits, the connexins. From these studies came the general assumption that global decreased connexin expression is correlated to tumor progression and increased cell proliferation. However, this assumption appeared more complicated by the fact that connexins may act also as pro-tumorigenic. Then, the concept that direct intercellular communication could be involved in cancer has been expanded to include new forms of intercellular communication such as tunneling nanotubes (TNTs) and exosomes. TNTs are intercellular bridges that allow free exchange of small molecules or even mitochondria depending on the presence of gap junctions. The majority of current research shows that such exchanges promote cancer progression by increasing resistance to hypoxia and chemotherapy. If exosomes are also involved in these mechanisms, more studies are needed to understand their precise role. Prostate cancer (PCa) represents a type of malignancy with one of the highest incidence rates worldwide. The precise role of these types of direct short-range intercellular communication has been considered in the progression of PCa. However, even though data are in favor of connexins playing a key role in PCa progression, a clear understanding of the role of TNTs and exosomes is needed to define their precise role in this malignancy. This review article summarizes the current view of the main mechanisms involved in short-range intercellular communication and their implications in cancer and delves into the biological, predictive and therapeutic role of connexins in PCa.
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Homeostatic control of pyramidal neuron firing rate involves a functional balance of feedforward excitation and feedback inhibition in neocortical circuits. Here, we reveal a dynamic scaling in vesicular excitatory (vesicular glutamate transporters VGLUT1 and VGLUT2) and inhibitory (vesicular inhibitory amino acid transporter VIAAT) transporter mRNA and synaptic protein expression in rat neocortical neuronal cultures, using a well established in vitro protocol to induce homeostatic plasticity. During the second and third week of synaptic differentiation, the predominant vesicular transporters expressed in neocortical neurons, VGLUT1 and VIAAT, are both dramatically upregulated. In mature cultures, VGLUT1 and VIAAT exhibit bidirectional and opposite regulation by prolonged activity changes. Endogenous coregulation during development and homeostatic scaling of the expression of the transporters in functionally differentiated cultures may serve to control vesicular glutamate and GABA filling and adjust functional presynaptic excitatory/inhibitory balance. Unexpectedly, hyperexcitation in differentiated cultures triggers a striking increase in VGLUT2 mRNA and synaptic protein, whereas decreased excitation reduces levels. VGLUT2 mRNA and protein are expressed in subsets of VGLUT1-encoded neocortical neurons that we identify in primary cultures and in neocortex in situ and in vivo. After prolonged hyperexcitation, downregulation of VGLUT1/synaptophysin intensity ratios at most synapses is observed, whereas a subset of VGLUT1-containing boutons selectively increase the expression of VGLUT2. Bidirectional and opposite regulation of VGLUT1 and VGLUT2 by activity may serve as positive or negative feedback regulators for cortical synaptic transmission. Intracortical VGLUT1/VGLUT2 coexpressing neurons have the capacity to independently modulate the level of expression of either transporter at discrete synapses and therefore may serve as a plastic interface between subcortical thalamic input (VGLUT2) and cortical output (VGLUT1) neurons.
Assuntos
Homeostase , Neocórtex/metabolismo , Proteína Vesicular 1 de Transporte de Glutamato/metabolismo , Proteína Vesicular 2 de Transporte de Glutamato/metabolismo , Proteínas Vesiculares de Transporte de Aminoácidos Inibidores/metabolismo , Envelhecimento , Animais , Animais Recém-Nascidos , Axônios/metabolismo , Células Cultivadas , Técnicas In Vitro , Neocórtex/citologia , Neocórtex/crescimento & desenvolvimento , Vias Neurais/citologia , Vias Neurais/crescimento & desenvolvimento , Vias Neurais/metabolismo , Neurônios/metabolismo , Células Piramidais/metabolismo , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Sinapses/metabolismo , Sinapses/fisiologia , Distribuição Tecidual , Regulação para Cima , Proteína Vesicular 1 de Transporte de Glutamato/genética , Proteína Vesicular 2 de Transporte de Glutamato/genéticaRESUMO
Male fertility is a highly controlled process that allows proliferation, meiosis and differentiation of male germ cells in the testis, final maturation in the epididymis and also requires functional male accessory glands: seminal vesicles, prostate and corpus cavernosum. In addition to classical endocrine and paracrine controls, mainly by gonadotropins LH and FSH and steroids, there is now strong evidence that all these processes are dependent upon the presence of homocellular or heterocellular junctions, including gap junctions and their specific connexins (Cxs), between the different cell types that structure the male reproductive tract. The present review is focused on the identification of Cxs, their distribution in the testis and in different structures of the male genital tract (epididymis, seminal vesicle, prostate, corpus cavernosum), their crucial role in the control of spermatogenesis and their implication in the function of the male accessory glands, including functional smooth muscle tone. Their potential dysfunctions in some testis (spermatogenic arrest, seminoma) and prostate (benign hyperplasia, adenocarcinoma) diseases and in the physiopathology of the human erectile function are also discussed.
Assuntos
Comunicação Celular , Conexinas/fisiologia , Junções Comunicantes/fisiologia , Espermatogênese , Testículo/metabolismo , Animais , Diferenciação Celular , Proliferação de Células , Fertilidade , Células Germinativas , Humanos , Masculino , Meiose , Próstata/metabolismo , Doenças Prostáticas/metabolismo , ReproduçãoRESUMO
Precise diagnosis of low and high grades of brain tumors permits determining therapeutical strategies. So far, diagnosis and prognosis of gliomas were based on histological and genetic criteria which need being completed by a panel of molecular markers. Highly distributed in brain, gap junction proteins, connexins, could be considered as markers of glioma progression as previous studies indicated that expression of a connexin type, connexin43 (Cx43), is inversely correlated to tumor grading. However, this assumption was weakened by the low number of glioma samples used. Taking advantage of tissue microarray technique, we pursued this analysis by studying in situ expression of Cx43 on 85 samples (37 grade IV, 18 grade III, 24 grade II, and 6 grades II to III). Our analysis confirmed the global diminution of Cx43 expression in glioblastomas that was observed in previous studies. However, this analysis brought new insights such as the following ones. First, the high number of samples permitted to show that more than 60% of glioblastomas still express Cx43. Second, no gradual decrease in Cx43 expression was observed between grades II and III, but Cx43 appeared to be a marker distinguishing oligodendrocytic and astrocytic grade III tumors. Third, independently from tumor grade, a Cx43 nuclear staining was detected in areas where leukocytes are present. In conclusion, our study emphasizes the importance of in situ immunohistochemical approaches by giving more precise insights in the subcellular localization of Cx43. It also emphasizes the necessity to carry out such analysis on a wide range of samples to circumvent the high glioma heterogeneity.
Assuntos
Biomarcadores Tumorais/metabolismo , Neoplasias Encefálicas/metabolismo , Conexina 43/metabolismo , Glioma/metabolismo , Adolescente , Adulto , Idoso , Western Blotting/métodos , Neoplasias Encefálicas/patologia , Núcleo Celular/metabolismo , Quimiotaxia de Leucócito , Feminino , Glioblastoma/metabolismo , Glioblastoma/patologia , Glioma/patologia , Humanos , Masculino , Pessoa de Meia-Idade , Gradação de Tumores , Proteínas de Neoplasias/metabolismo , Análise Serial de Tecidos/métodos , Adulto JovemRESUMO
Formation of metastases negatively impacts the survival prognosis of cancer patients. Globally, if the various steps involved in their formation are relatively well identified, the molecular mechanisms responsible for the emergence of invasive cancer cells are still incompletely resolved. Elucidating what are the mechanisms that allow cancer cells to evade from the tumor is a crucial point since it is the first step of the metastatic potential of a solid tumor. In order to be invasive, cancer cells have to undergo transformations such as down-regulation of cell-cell adhesions, modification of cell-matrix adhesions and acquisition of proteolytic properties. These transformations are accompanied by the capacity to "activate" stromal cells, which may favor the motility of the invasive cells through the extracellular matrix. Since modulation of gap junctional intercellular communication is known to be involved in cancer, we were interested to consider whether these different transformations necessary for the acquisition of invasive phenotype are related with gap junctions and their structural proteins, the connexins. In this review, emerging roles of connexins and gap junctions in the process of tissue invasion are proposed.
Assuntos
Conexinas/metabolismo , Transição Epitelial-Mesenquimal , Junções Comunicantes/metabolismo , Neoplasias/patologia , Animais , Adesão Celular , Humanos , Invasividade Neoplásica , Neoplasias/metabolismo , ProteóliseRESUMO
For decades, cancer was associated with gap-junction defects. However, more recently it appeared that the gap junction proteins (connexins) could be re-expressed and participate to cancer cell dissemination during the late stages of tumor progression. Since primary tumors of prostate cancer (PCa) are known to be connexin deficient, it was interesting to verify whether their bone-targeted metastatic behaviour could be influenced by the re-expression of the connexin type (connexin43) which is originally present in prostate tissue and highly expressed in bone where it participates to the differentiation of osteoblastic cells. Thus, we investigated the effect of the increased Cx43 expression, by retroviral infection, on the metastatic behaviour of two well-characterized cell lines (PC-3 and LNCaP) representing different stages of PCa progression. It appeared that Cx43 differently behaved in those cell lines and induced different phenotypes. In LNCaP, Cx43 was functional, localized at the plasma membrane and its high expression was correlated with a more aggressive phenotype both in vitro and in vivo. In particular, those Cx43-expressing LNCaP cells exhibited a high incidence of osteolytic metastases generated by bone xenografts in mice. Interestingly, LNCaP cells were also able to decrease the proliferation of cocultured osteoblastic cells. In contrast, the increased expression of Cx43 in PC-3 cells led to an unfunctional, cytoplasmic localization of the protein and was correlated with a reduction of proliferation, adhesion and invasion of the cells. In conclusion, the localization and the functionality of Cx43 may govern the ability of PCa cells to metastasize in bones.
Assuntos
Neoplasias Ósseas/secundário , Conexina 43/fisiologia , Junções Comunicantes/fisiologia , Neoplasias da Próstata/patologia , Western Blotting , Neoplasias Ósseas/fisiopatologia , Adesão Celular , Linhagem Celular Tumoral , Proliferação de Células , Técnicas de Cocultura , Humanos , Imuno-Histoquímica , Masculino , Neoplasias da Próstata/fisiopatologia , Reação em Cadeia da Polimerase em Tempo RealRESUMO
Cancer was one of the first pathologies to be associated with gap-junction defect. Despite the evidence accumulated over the last 40-year period, the molecular involvement of gap junctions and their structural proteins (connexins) in cancer has not been elucidated. The lack of a satisfying explanation may come from the complexity of the disease, evolving through various stages during tumor progression, with cancer cells exhibiting different phenotypes. Here, the question of the involvement of gap junctions has been readdressed by considering the connexin expression/function level at different fundamental stages of carcinogenesis (cell proliferation, cell invasion, and cancer cell dissemination). By performing this analysis, it becomes clear that gap junctions are probably differently involved, depending on the stage of the cancer progression considered. In particular, the most recent data suggest that connexins may act on cell growth by controlling gene expression through a variety of processes (independent of or dependent on the gap-junctional communication capacity). During invasion, connexins have been demonstrated to enhance adherence of cancer cells to the stroma, migration, and probably their dissemination by establishing communication with the endothelial barrier. All these data present a complex picture of connexins in various functions, depending on the cell phenotype.
Assuntos
Junções Comunicantes/metabolismo , Junções Comunicantes/fisiologia , Neoplasias/metabolismo , Comunicação Celular/fisiologia , Conexinas/metabolismo , Conexinas/fisiologia , Regulação Neoplásica da Expressão Gênica/fisiologia , Neoplasias/fisiopatologiaRESUMO
System A transporters SNAT1 and SNAT2 mediate uptake of neutral alpha-amino acids (e.g. glutamine, alanine, and proline) and are expressed in central neurons. We tested the hypothesis that SNAT2 is required to support neurotransmitter glutamate synthesis by examining spontaneous excitatory activity after inducing or repressing SNAT2 expression for prolonged periods. We stimulated de novo synthesis of SNAT2 mRNA and increased SNAT2 mRNA stability and total SNAT2 protein and functional activity, whereas SNAT1 expression was unaffected. Increased endogenous SNAT2 expression did not affect spontaneous excitatory action-potential frequency over control. Long term glutamine exposure strongly repressed SNAT2 expression but increased excitatory action-potential frequency. Quantal size was not altered following SNAT2 induction or repression. These results suggest that spontaneous glutamatergic transmission in pyramidal neurons does not rely on SNAT2. To our surprise, repression of SNAT2 activity was not limited to System A substrates. Taurine, gamma-aminobutyric acid, and beta-alanine (substrates of the SLC6 gamma-aminobutyric acid transporter family) repressed SNAT2 expression more potently (10x) than did System A substrates; however, the responses to System A substrates were more rapid. Since ATF4 (activating transcription factor 4) and CCAAT/enhancer-binding protein are known to bind to an amino acid response element within the SNAT2 promoter and mediate induction of SNAT2 in peripheral cell lines, we tested whether either factor was similarly induced by amino acid deprivation in neurons. We found that glutamine and taurine repressed the induction of both transcription factors. Our data revealed that SNAT2 expression is constitutively low in neurons under physiological conditions but potently induced, together with the taurine transporter TauT, in response to depletion of neutral amino acids.
Assuntos
Sistemas de Transporte de Aminoácidos/fisiologia , Proteínas da Membrana Plasmática de Transporte de GABA/metabolismo , Regulação da Expressão Gênica , Glutamina/metabolismo , Neocórtex/citologia , Neurônios/metabolismo , Sistema A de Transporte de Aminoácidos , Sistemas de Transporte de Aminoácidos/metabolismo , Animais , Células Cultivadas , Relação Dose-Resposta a Droga , Eletrofisiologia , Oócitos/metabolismo , Isoformas de Proteínas , Ratos , Xenopus laevisRESUMO
Gap junctional intercellular communication (GJIC) permits coordinated cellular activities during developmental and differentiation processes. In bone, the involvement of the gap junctional protein, connexin-43 (Cx43), and of GJIC in osteoblastic differentiation and mineralization of the extracellular matrix has been previously demonstrated. Former studies have shown that endothelin-1 (ET-1) was also implicated in the control of osteoblastic proliferation and differentiation. However, depending on the cellular models, ET-1 has been shown to decrease or increase osteoblastic differentiation markers. As no data were available on the ET-1 effect on GJIC and Cx43 expression in osteoblastic cells, we analyzed here the possible crosstalk between Cx43 and ET-1 in a human cell line (hFOB 1.19) which displays different Cx43 expression levels and phenotypes when cultured at 33.5 or 39 degrees C. The presence of ET-1 (10(-8) M) for 2-12 days of culture did not significantly alter the proliferation rate of hFOB cells whatever their phenotype. In contrast, ET-1 induced a differential inhibitory effect on the biochemical differentiation markers (alkaline phosphatase activity and osteocalcin expression) with a significant reduction in the differentiated phenotype at 39 degrees C, whereas no effects were measured at 33.5 degrees C. The inhibitory effect was linked to a decrease of GJIC and of Cx43 both at transcriptional and protein levels. Altogether, our results suggest that Cx43 expression level could influence the action of ET-1 on human osteoblastic cell differentiation. Our data also indicate that the gap junctional protein could play a pivotal role in the response of osteoblasts to mitogenic factors implicated in bone pathologies.
Assuntos
Diferenciação Celular/efeitos dos fármacos , Conexina 43/metabolismo , Endotelina-1/farmacologia , Osteoblastos/citologia , Osteoblastos/metabolismo , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Junções Comunicantes/efeitos dos fármacos , Junções Comunicantes/metabolismo , Humanos , Osteoblastos/efeitos dos fármacos , Receptor de Endotelina A/metabolismoRESUMO
During bone remodeling, osteoblastic (OB) cells have a central role leading to the production of extracellular matrix and its subsequent mineralization. As revealed by human physiopathologies, the OB differentiation process is essential for the control of calcium metabolism and normal bone formation. Moreover, accumulating data in the field of bone development suggest that connexin 43 (Cx43)-mediated gap junctional communication plays an important role in OB differentiation and function. Since Ca(2+) has a central role in OB physiology, the aim of the present study was to investigate the hypothetical involvement of Cx43 in OB calcium homeostasis. We performed measurements of intracellular calcium activity ([Ca(2+)]( i )) by a cytofluorimetric method using Fluo-4 as a calcium indicator and endothelin-1 (ET-1) as a physiological calcium-mobilizing factor on cultured OB cells isolated from calvaria of Cx43(+/-) and Cx43(+/+) mice. Partial deletion of the Cx43 gene induced a significant decrease in the [Ca(2+)]( i ) rise elicited by ET-1. This reduction was not correlated to a decrease or a modification of ET receptor subtype expression as assessed by real-time reverse-transcription polymerase chain reaction. Pharmacological investigations led us to demonstrate that the significant difference in [Ca(2+)]( i ) peak amplitude during the ET-1 action was associated with decreased calcium influx involving L-type voltage-sensitive calcium channels, whereas calcium release from intracellular stores and implication of phospholipase C were not affected by the reduced expression of Cx43. In conclusion, our data demonstrate for the first time that the Cx43 level of expression and/or function is able to modulate the [Ca(2+)]( i ) mobilization in OB cells.
Assuntos
Cálcio/metabolismo , Conexina 43/fisiologia , Endotelina-1/farmacologia , Osteoblastos/fisiologia , Crânio/metabolismo , Animais , Animais Recém-Nascidos , Sinalização do Cálcio , Comunicação Celular , Células Cultivadas , Conexina 43/genética , Junções Comunicantes/metabolismo , Heterozigoto , Humanos , Camundongos , Camundongos Knockout , Camundongos Transgênicos , Osteoblastos/citologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptor de Endotelina A/metabolismo , Receptor de Endotelina B/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Crânio/citologia , Fosfolipases Tipo C/metabolismoRESUMO
The gap junction proteins, connexins (Cxs), are present in the testis, and among them, Cx43 play an essential role in spermatogenesis. In the present study, we investigated the testicular expression and regulation of another Cx, Cx33, previously described as a negative regulator of gap junction communication. Cx33 mRNA was present in testis and undetectable in heart, liver, ovary, and uterus. In the mature testis, Cx33 was specifically immunolocalized in the basal compartment of the seminiferous tubules, whereas Cx43 was present in both seminiferous tubule and interstitial compartments. During stages IX and X of spermatogenesis, characterized by Sertoli cell phagocytosis of residual bodies, Cx43 was poorly expressed within seminiferous tubules, while Cx33 signal was strong. To evaluate the role of phagocytosis in the control of Cx33 and Cx43 expression, the effect of LPS was analyzed in the Sertoli cell line 42GPA9. We show herein that phagocytosis activation by LPS concomitantly stimulated Cx33 and inhibited Cx43 mRNA levels. These effects appear to have been mediated through IL-1alpha, because the exposure of Sertoli cells to the IL-1 receptor antagonist partly reversed these effects. IL-1alpha enhanced and reduced, respectively, the levels of Cx33 and Cx43 mRNA in a time- and dose-dependent manner. These data reveal that Cx33 and Cx43 genes are controlled differently within the testis and suggest that these two Cxs may exert opposite and complementary effects on spermatogenesis.