RESUMO
p53 is a much studied transcription factor which has a key role in the maintenance of genetic stability. It belongs to a larger family of genes including two other highly related proteins, p63 and p73. The p53 pathway has a vital role in the prevention of cancer formation and is ubiquitously lost in a high percentage of human cancers. In 60% of cancer cases this occurs via p53 gene mutation. In the remaining cancers expressing a WTp53 gene, loss of cell signalling upstream or downstream of p53 are responsible for the inactivation of the p53 pathway. It has recently been described that the p53 gene encodes for nine different p53 isoforms, whereas the p63 and p73 genes encode for at least other 6 and 29, respectively. This finding may have a profound impact on our comprehension of p53 tumour suppressor activity. Studies in several tumour types have shown abnormal expression of these protein isoforms. Hence, better understanding of p53 tumour suppressor activity and the interaction between p53 family members and their isoforms is likely to bring us closer to cancer therapy. Therapeutic manipulation of the p53 pathway is therefore a highly promising field and already the focus of extensive investigation. Many strategies are being developed to either restore inactive/suppressed wild-type p53 (WTp53) or reverse the p53 mutant phenotype into WTp53. As p53 pathway inactivation is a common denominator to all cancers, it is highly expected that these therapies will be able to target a broad range of cancers and will allow for more specific targeting of cancer cells, avoiding collateral damage to normal tissue.
Assuntos
Antineoplásicos/uso terapêutico , Mutação/genética , Neoplasias/diagnóstico , Neoplasias/terapia , Proteína Supressora de Tumor p53/genética , Proteínas Supressoras de Tumor/genética , Humanos , Neoplasias/genética , Prognóstico , Isoformas de ProteínasRESUMO
In the malaria vector Anopheles gambiae, two point mutations in the acetylcholinesterase (ace-1R) and the sodium channel (kdrR) genes confer resistance to organophosphate/carbamate and pyrethroid insecticides, respectively. The mechanisms of compensation that recover the functional alterations associated with these mutations and their role in the modulation of insecticide efficacy are unknown. Using multidisciplinary approaches adapted to neurons isolated from resistant Anopheles gambiae AcerKis and KdrKis strains together with larval bioassays, we demonstrate that nAChRs, and the intracellular calcium concentration represent the key components of an adaptation strategy ensuring neuronal functions maintenance. In AcerKis neurons, the increased effect of acetylcholine related to the reduced acetylcholinesterase activity is compensated by expressing higher density of nAChRs permeable to calcium. In KdrKis neurons, changes in the biophysical properties of the L1014F mutant sodium channel, leading to enhance overlap between activation and inactivation relationships, diminish the resting membrane potential and reduce the fraction of calcium channels available involved in acetylcholine release. Together with the lower intracellular basal calcium concentration observed, these factors increase nAChRs sensitivity to maintain the effect of low concentration of acetylcholine. These results explain the opposite effects of the insecticide clothianidin observed in AcerKis and KdrKis neurons in vitro and in vivo.
Assuntos
Anopheles/efeitos dos fármacos , Anopheles/genética , Inseticidas/farmacologia , Mosquitos Vetores/efeitos dos fármacos , Mosquitos Vetores/genética , Acetilcolina/farmacologia , Acetilcolinesterase/genética , Animais , Anopheles/metabolismo , Cálcio/metabolismo , Canais de Cálcio/metabolismo , Feminino , Genes de Insetos , Humanos , Técnicas In Vitro , Resistência a Inseticidas/genética , Malária/transmissão , Controle de Mosquitos/métodos , Mosquitos Vetores/metabolismo , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Técnicas de Patch-Clamp , Mutação Puntual , Receptores Nicotínicos/metabolismo , Canais de Sódio/genéticaRESUMO
The insect repellent IR3535 is one of the important alternative in the fight against mosquito-borne disease such as malaria, dengue, chikungunya, yellow fever and Zika. Using a multidisciplinary approach, we propose the development of an innovative insecticide-based vector control strategy using an unexplored property of IR3535. We have demonstrated that in insect neurosecretory cells, very low concentration of IR3535 induces intracellular calcium rise through cellular mechanisms involving orthosteric/allosteric sites of the M1-muscarinic receptor subtype, G protein ßγ subunits, background potassium channel inhibition generating depolarization, which induces voltage-gated calcium channel activation. The resulting internal calcium concentration elevation increases nicotinic receptor sensitivity to the neonicotinoid insecticide thiacloprid. The synergistic interaction between IR3535 and thiacloprid contributes to significantly increase the efficacy of the treatment while reducing concentrations. In this context, IR3535, used as a synergistic agent, seems to promise a new approach in the optimization of the integrated vector management for vector control.
Assuntos
Controle de Insetos , Proteínas de Insetos/metabolismo , Insetos Vetores/metabolismo , Inseticidas/farmacologia , Periplaneta/metabolismo , Receptores Muscarínicos/metabolismo , beta-Alanina/análogos & derivados , Animais , Masculino , beta-Alanina/farmacologiaRESUMO
Migration stimulating factor (MSF) is a truncated oncofetal fibronectin isoform expressed by fetal and tumor-associated cells. MSF mRNA is distinguished from other fibronectin isoforms by its size (2.1 kb) and the inclusion of a specific intronic sequence at its 3' end. Initial Northern blot analysis with a MSF-specific probe indicated the presence of this 2.1-kb transcript and an additional unexpected 5.9-kb RNA present in both MSF-secreting (fetal) and nonsecreting (adult) fibroblasts. Our investigations into the nature of these transcripts and their relationship to MSF protein secretion revealed that the 5.9-kb mRNA is a second MSF-encoding transcript. Both these mRNAs have identical coding sequence and differ only in the length of their intron-derived 3'-untranslated region (UTR). The 5.9-kb MSF mRNA is retained in the nucleus whereas the 2.1-kb mRNA is not. MSF-secreting fetal fibroblasts have significantly lower nuclear levels of the 5.9-kb mRNA and correspondingly higher cytoplasmic levels of the 2.1-kb transcript than their nonsecreting adult counterparts. Adult fibroblasts induced to secrete MSF by treatment with transforming growth factor-beta1 displayed similar changes in their respective levels of MSF mRNA, but not those of a control gene. When cloned downstream of a reporter gene, only the longer 3'-UTR retained coding sequence within the nucleus. We conclude that expression of MSF protein is regulated by 3'-UTR truncation of the 5.9-kb nuclear-sequestered "precursor" MSF mRNA and nuclear export of mature 2.1-kb message. Inducible 3'-UTR processing represents a novel regulatory mechanism involved in cancer pathogenesis that may open new avenues for therapeutic gene delivery.
Assuntos
Regiões 3' não Traduzidas/metabolismo , Citocinas/biossíntese , Precursores de RNA/metabolismo , Regiões 3' não Traduzidas/genética , Adulto , Sequência de Aminoácidos , Linhagem Celular , Núcleo Celular/metabolismo , Citocinas/genética , Citocinas/metabolismo , Fibroblastos/citologia , Fibroblastos/metabolismo , Fibroblastos/fisiologia , Fibronectinas , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Dados de Sequência Molecular , Precursores de RNA/genética , Frações Subcelulares/metabolismoRESUMO
Interleukin-1 receptor antagonist (IL-1ra) has been shown to play a crucial role in the prevention of various inflammatory diseases. There is also convincing evidence that IL-1ra is able to counteract inflammatory effects of IL-1 members implicated in insulin resistance and diabetes. However, the use of knock-out animal models provides evidence to the contrary and the role of IL-1ra in obesity-linked anomalies remains controversial. This minireview gets an insight into recent findings on the implication of IL-1ra and its gene polymorphism in diabetes and obesity, discusses the potential dual effects of IL-1ra observed in different models, and comments on future directions.
Assuntos
Diabetes Mellitus/metabolismo , Resistência à Insulina , Proteína Antagonista do Receptor de Interleucina 1/metabolismo , Interleucina-1/antagonistas & inibidores , Obesidade/metabolismo , Animais , Diabetes Mellitus/genética , Modelos Animais de Doenças , Humanos , Inflamação/imunologia , Inflamação/metabolismo , Inflamação/prevenção & controle , Resistência à Insulina/genética , Proteína Antagonista do Receptor de Interleucina 1/genética , Proteína Antagonista do Receptor de Interleucina 1/uso terapêutico , Interleucina-1/genética , Interleucina-1/metabolismo , Camundongos , Camundongos Knockout , Obesidade/genética , Polimorfismo GenéticoRESUMO
Insulin regulation of hepatic gene transcription is a vital component of glucose homeostasis. Understanding the molecular regulationof thisprocess aids the searchfor the defect(s) that promotesinsulin-resistant states, such asdiabetesmellitus. We havepreviously shownthat the insulin regulationof hepatic IGF-binding protein-1 (IGFBP1) expression requiresthe signalling proteins phosphatidylinositol 3-kinase (PI 3-kinase) and mammalian target of rapamycin (mTOR). In this report, we demonstrate that activation of the mTOR pathway, without activation of its upstream regulator PI 3-kinase, reduces IGFBP1 expression. Therefore, mTOR activation is sufficient to mimic insulin regulation of this gene. However, longer exposure (>3 h) of cells to insulin reduces the importance of this pathway in insulin regulation of the gene, suggesting a temporal switch in signalling mechanisms linking insulin action to the IGFBP1 gene promoter. In contrast, the activation of PI 3-kinase is required for insulin regulation of IGFBP1 under all conditions tested. Therefore, an mTOR-independent, PI 3-kinase-dependent pathway becomes more important in IGFBP1 regulation after long exposure to insulin. This is a novel concept in insulin regulation of gene expression and demonstrates the importance of temporal analysis of signalling processes.
Assuntos
Proteína 1 de Ligação a Fator de Crescimento Semelhante à Insulina/metabolismo , Insulina/metabolismo , Fígado/metabolismo , Animais , Cicloeximida/farmacologia , Regulação para Baixo , Regulação da Expressão Gênica , Fígado/efeitos dos fármacos , Modelos Biológicos , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Quinases/metabolismo , Ratos , Transdução de Sinais , Sirolimo/farmacologia , Serina-Treonina Quinases TOR , Fatores de Tempo , Células Tumorais CultivadasRESUMO
Zinc-α2-glycoprotein (ZAG) is a new adipokine whose gene expression is downregulated in obese patients. We recently reported ZAG expression in breast tumor or healthy breast tissue and detected this expression at high levels in ductal carcinoma and in normal epithelial adjacent tissue but not in normal tissue of healthy women. In the present study, we used two human breast tumor cell lines (MCF-7 and MDA-MB231) and one fibrocystic breast cell line (MCF10a) to examine whether recombinant ZAG has an effect on proliferative/apoptotic response in breast cancer cell lines. ZAG seemed to exert a proliferative effect on breast cancer cell proliferation [+11 to 27% in MCF-7 with (ZAG) = 5-20 µg/ml; +13% in MDA-MB-231 with (ZAG) = 5 µg/ml] and, on the contrary, an anti-proliferative effect in the fibrocystic breast cell line [-5 to -8% in MCF-10a with (ZAG) = 5-10 µg/ml]. ZAG was able to modulate gene and protein expression involved in the apoptotic response. However, further studies are required to fully elucidate the effects of ZAG on the proliferation of mammary cells.
Assuntos
Neoplasias da Mama/genética , Neoplasias da Mama/metabolismo , Proteínas de Plasma Seminal/genética , Proteínas de Plasma Seminal/metabolismo , Apoptose/genética , Neoplasias da Mama/patologia , Processos de Crescimento Celular/genética , Linhagem Celular , Linhagem Celular Tumoral , Feminino , Expressão Gênica , Humanos , Células MCF-7 , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Glicoproteína Zn-alfa-2RESUMO
Obesity is associated with an increased risk of breast cancer in postmenopausal women. Accumulating evidence suggests that adipose tissue, which is an endocrine organ producing a large range of factors, may interfere with breast cancer development. Leptin and adiponectin are two major adipocyte-secreted hormones. The pro-carcinogenic effect of leptin and conversely, the anti-carcinogenic effect of adiponectin result from two main mechanisms: a modulation in the signalling pathways involved in proliferation process and a subtle regulation of the apoptotic response. This review provides insight into recent findings on the molecular mechanisms of leptin and adiponectin in mammary tumours, and discusses the potential interplay between these two adipokines in breast cancer.
Assuntos
Adiponectina/fisiologia , Tecido Adiposo/metabolismo , Neoplasias da Mama/etiologia , Leptina/fisiologia , Obesidade/complicações , Adiponectina/farmacologia , Animais , Comunicação Celular/fisiologia , Feminino , Humanos , Leptina/farmacologia , Camundongos , Receptores para Leptina/fisiologia , Células Tumorais CultivadasRESUMO
p53 gene expresses several protein isoforms modulating p53-mediated responses through regulation of gene expression. Here, we identify a novel p53 isoform, Δ160p53, lacking the first 159 residues. By knockdown experiments and site-directed mutagenesis, we show that Δ160p53 is encoded by Δ133p53 transcript using ATG160 as translational initiation site. This hypothesis is supported by endogenous expression of Δ160p53 in U2OS, T47D and K562 cells, the latter ones carrying a premature stop codon that impairs p53 and Δ133p53 protein expression but not the one of Δ160p53. Overall, these results show that the Δ133p53 transcript generates two different p53 isoforms, Δ133p53 and Δ160p53.
Assuntos
Proteína Supressora de Tumor p53/química , Proteína Supressora de Tumor p53/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Cães , Técnicas de Silenciamento de Genes , Humanos , Células K562 , Camundongos , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , Isoformas de Proteínas/química , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , RNA Mensageiro/genética , Ratos , Deleção de Sequência , Proteína Supressora de Tumor p53/metabolismoRESUMO
Obesity is associated with an increased risk of breast cancer. interleukin-1 (IL-1), a pro-inflammatory cytokine secreted by adipose tissue, is involved in breast cancer development. There is also convincing evidence that other adipocytokines including leptin not only have a role in haematopoiesis, reproduction and immunity but are also growth factors in cancer. Therefore, IL-1 family and leptin family are adipocytokines which could represent a major link between obesity and breast cancer progression. This minireview provides insight into recent findings on the prognostic significance of IL-1 and leptin in mammary tumours, and discusses the potential interplay between IL-1 family members and adipocyte-derived hormones in breast cancer.