RESUMEN

The transcription factor ΔNp63 plays a pivotal role in maintaining the integrity of stratified epithelial tissues by regulating the expression of distinct target genes involved in lineage specification, cell stemness, cell proliferation and differentiation. Here, we identified the ABC transporter subfamily member ABCC1 as a novel ΔNp63 target gene. We found that in immortalized human keratinocytes and in squamous cell carcinoma (SCC) cells, ∆Np63 induces the expression of ABCC1 by physically occupying a p63-binding site (p63 BS) located in the first intron of the ABCC1 gene locus. In cutaneous SCC and during the activation of the keratinocyte differentiation program, ∆Np63 and ABCC1 levels are positively correlated raising the possibility that ABCC1 might be involved in the regulation of the proliferative/differentiative capabilities of squamous tissue. However, we did not find any gross alteration in the structure and morphology of the epidermis in humanized hABCC1 knock-out mice. Conversely, we found that the genetic ablation of ABCC1 led to a marked reduction in inflammation-mediated proliferation of keratinocytes, suggesting that ABCC1 might be involved in the regulation of keratinocyte proliferation upon inflammatory/proliferative signals. In line with these observations, we found a significant increase in ABCC1 expression in squamous cell carcinomas (SCCs), a tumor type characterized by keratinocyte hyper-proliferation and a pro-inflammatory tumor microenvironment. Collectively, these data uncover ABCC1 as an additional ∆Np63 target gene potentially involved in those skin diseases characterized by dysregulation of proliferation/differentiation balance.

Asunto(s)

Carcinoma de Células Escamosas , Proliferación Celular , Regulación Neoplásica de la Expresión Génica , Queratinocitos , Proteínas Asociadas a Resistencia a Múltiples Medicamentos , Neoplasias Cutáneas , Factores de Transcripción , Proteínas Supresoras de Tumor , Humanos , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/metabolismo , Carcinoma de Células Escamosas/patología , Proteínas Asociadas a Resistencia a Múltiples Medicamentos/genética , Proteínas Asociadas a Resistencia a Múltiples Medicamentos/metabolismo , Animales , Queratinocitos/metabolismo , Queratinocitos/patología , Ratones , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Neoplasias Cutáneas/genética , Neoplasias Cutáneas/patología , Neoplasias Cutáneas/metabolismo , Proteínas Supresoras de Tumor/genética , Proteínas Supresoras de Tumor/metabolismo , Proliferación Celular/genética , Diferenciación Celular/genética , Ratones Noqueados , Transactivadores/genética , Transactivadores/metabolismo , Línea Celular Tumoral

RESUMEN

Defects in achieving a fully differentiated state and aberrant expression of genes and microRNAs (miRs) involved in differentiation are common to virtually all tumor types. Here, we demonstrate that the zinc finger transcription factor ZNF281/Zfp281 is down-regulated during epithelial, muscle, and granulocytic differentiation in vitro. The expression of this gene is absent in terminally differentiated human tissues, in contrast to the elevated expression in proliferating/differentiating ones. Analysis of the 3'UTR of ZNF281/Zfp281 revealed the presence of numerous previously undescribed miR binding sites that were proved to be functional for miR-mediated post-transcriptional regulation. Many of these miRs are involved in differentiation pathways of distinct cell lineages. Of interest, ZNF281/Zfp281 is able to inhibit muscle differentiation promoted by miR-1, of which ZNF281/Zfp281 is a direct target. These data suggest that down-regulation of ZNF281/Zfp281 during differentiation in various cell types may occur through specific miRs whose expression is tissue-restricted. In addition, we found that in rhabdomyosarcoma and leiomyosarcoma tumors, the expression of ZNF281/Zfp281 is significantly higher compared with normal counterparts. We extended our analysis to other human soft tissue sarcomas, in which the expression of ZNF281 is associated with a worse prognosis. In summary, we highlight here a new role of ZNF281/Zfp281 in counteracting muscle differentiation; its down-regulation is at least in part mediated by miR-1. The elevated expression of ZNF281/Zfp281 in soft tissue sarcomas warrants further analysis for its possible exploitation as a prognostic marker in this class of tumors.

Asunto(s)

MicroARNs/metabolismo , Desarrollo de Músculos/genética , Proteínas Represoras/metabolismo , Sarcoma/metabolismo , Factores de Transcripción/metabolismo , Animales , Línea Celular Tumoral , Perfilación de la Expresión Génica , Humanos , Estimación de Kaplan-Meier , Ratones , MicroARNs/genética , Mioblastos/metabolismo , Células 3T3 NIH , Pronóstico , Procesamiento Proteico-Postraduccional/genética , Proteínas Represoras/genética , Sarcoma/genética , Sarcoma/mortalidad , Factores de Transcripción/genética

RESUMEN

Efficient repair of DNA double-strand breaks (DSBs) is of critical importance for cell survival. Although non-homologous end joining (NHEJ) is the most used DSBs repair pathway in the cells, how NHEJ factors are sequentially recruited to damaged chromatin remains unclear. Here, we identify a novel role for the zinc-finger protein ZNF281 in participating in the ordered recruitment of the NHEJ repair factor XRCC4 at damage sites. ZNF281 is recruited to DNA lesions within seconds after DNA damage through a mechanism dependent on its DNA binding domain and, at least in part, on poly-ADP ribose polymerase (PARP) activity. ZNF281 binds XRCC4 through its zinc-finger domain and facilitates its recruitment to damaged sites. Consequently, depletion of ZNF281 impairs the efficiency of the NHEJ repair pathway and decreases cell viability upon DNA damage. Survival analyses from datasets of commonly occurring human cancers show that higher levels of ZNF281 correlate with poor prognosis of patients treated with DNA-damaging therapies. Thus, our results define a late ZNF281-dependent regulatory step of NHEJ complex assembly at DNA lesions and suggest additional possibilities for cancer patients' stratification and for the development of personalised therapeutic strategies.

Asunto(s)

Roturas del ADN de Doble Cadena , Reparación del ADN por Unión de Extremidades , Neoplasias/genética , Neoplasias/patología , Proteínas Represoras/metabolismo , Sistemas CRISPR-Cas , Línea Celular Tumoral , Proteínas de Unión al ADN/metabolismo , Bases de Datos Genéticas , Humanos , Neoplasias/metabolismo , Poli(ADP-Ribosa) Polimerasa-1/genética , Poli(ADP-Ribosa) Polimerasa-1/metabolismo , Pronóstico , Proteínas Represoras/genética , Tasa de Supervivencia

RESUMEN

Mutations in the TP53 gene and microenvironmentally driven activation of hypoxia-inducible factor-1 (HIF-1) typically occur in later stages of tumorigenesis. An ongoing challenge is the identification of molecular determinants of advanced cancer pathogenesis to design alternative last-line therapeutic options. Here, we report that p53 mutants influence the tumor microenvironment by cooperating with HIF-1 to promote cancer progression. We demonstrate that in non-small cell lung cancer (NSCLC), p53 mutants exert a gain-of-function (GOF) effect on HIF-1, thus regulating a selective gene expression signature involved in protumorigenic functions. Hypoxia-mediated activation of HIF-1 leads to the formation of a p53 mutant/HIF-1 complex that physically binds the SWI/SNF chromatin remodeling complex, promoting expression of a selective subset of hypoxia-responsive genes. Depletion of p53 mutants impairs the HIF-mediated up-regulation of extracellular matrix (ECM) components, including type VIIa1 collagen and laminin-γ2, thus affecting tumorigenic potential of NSCLC cells in vitro and in mouse models in vivo. Analysis of surgically resected human NSCLC revealed that expression of this ECM gene signature was highly correlated with hypoxic tumors exclusively in patients carrying p53 mutations and was associated with poor prognosis. Our data reveal a GOF effect of p53 mutants in hypoxic tumors and suggest synergistic activities of p53 and HIF-1. These findings have important implications for cancer progression and might provide innovative last-line treatment options for advanced NSCLC.

Asunto(s)

Carcinoma de Pulmón de Células no Pequeñas/genética , Factor 1 Inducible por Hipoxia/genética , Neoplasias Pulmonares/genética , Proteína p53 Supresora de Tumor/metabolismo , Animales , Carcinoma de Pulmón de Células no Pequeñas/metabolismo , Carcinoma de Pulmón de Células no Pequeñas/patología , Hipoxia de la Célula/genética , Línea Celular Tumoral , Matriz Extracelular , Genes p53 , Xenoinjertos , Humanos , Factor 1 Inducible por Hipoxia/metabolismo , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patología , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Mutación , Activación Transcripcional , Microambiente Tumoral , Proteína p53 Supresora de Tumor/genética

RESUMEN

Derangement of cellular differentiation because of mutation or inappropriate expression of specific genes is a common feature in tumors. Here, we show that the expression of ZNF281, a zinc finger factor involved in several cellular processes, decreases during terminal differentiation of murine cortical neurons and in retinoic acid-induced differentiation of neuroblastoma (NB) cells. The ectopic expression of ZNF281 inhibits the neuronal differentiation of murine cortical neurons and NB cells, whereas its silencing causes the opposite effect. Furthermore, TAp73 inhibits the expression of ZNF281 through miR34a. Conversely, MYCN promotes the expression of ZNF281 at least in part by inhibiting miR34a. These findings imply a functional network that includes p73, MYCN, and ZNF281 in NB cells, where ZNF281 acts by negatively affecting neuronal differentiation. Array analysis of NB cells silenced for ZNF281 expression identified GDNF and NRP2 as two transcriptional targets inhibited by ZNF281. Binding of ZNF281 to the promoters of these genes suggests a direct mechanism of repression. Bioinformatic analysis of NB datasets indicates that ZNF281 expression is higher in aggressive, undifferentiated stage 4 than in localized stage 1 tumors supporting a central role of ZNF281 in affecting the differentiation of NB. Furthermore, patients with NB with high expression of ZNF281 have a poor clinical outcome compared with low-expressors. These observations suggest that ZNF281 is a controller of neuronal differentiation that should be evaluated as a prognostic marker in NB.

Asunto(s)

Biomarcadores de Tumor/biosíntesis , Diferenciación Celular , Proteínas de Neoplasias/biosíntesis , Neuroblastoma/metabolismo , Neuronas/metabolismo , Transactivadores/biosíntesis , Factores de Transcripción/biosíntesis , Animales , Biomarcadores de Tumor/genética , Línea Celular Tumoral , Regulación Neoplásica de la Expresión Génica , Humanos , Ratones , Proteínas de Neoplasias/genética , Neuroblastoma/diagnóstico , Neuroblastoma/genética , Neuroblastoma/patología , Neuronas/patología , Pronóstico , Proteínas Represoras , Transactivadores/genética , Factores de Transcripción/genética

RESUMEN

INTRODUCTION: Major depressive disorder (MDD) and osteoporosis are two common disorders with high morbidity and mortality rates. Conflicting data have found associations between MDD and low bone mineral density (BMD) or osteoporosis, although causative factors are still unclear. A pilot study was designed with the aim to assess the relationship between MDD and BMD in postmenopausal women with MDD compared to healthy volunteers. We hypothesized that attachment style (AS) mediated this relationship. METHODS: The sample was made of 101 postmenopausal women, 49 with MDD and 52 age-matched healthy volunteers. Structured clinical interview and Beck Depression Inventory (BDI) were performed to assesse MDD. AS was evaluated using the Relationship Questionnaire (RQ). BMD was measured by dual energy X-ray absorptiometry. RESULTS: The univariate analysis showed that women with MDD had lower BMD values as compared to healthy volunteers. In the regression models MDD diagnosis and BDI score were not significant predictors of low BMD. The "preoccupied" pattern of insecure AS was a significant, independent predictor of decreased BMD in all skeletal sites: lumbar spine (p=0.008), femoral neck (p=0.011), total hip (p=0.002). CONCLUSIONS: This is the first study exploring the relationship between AS, MDD and BMD. Our results support the link between MDD and low BMD. We found that insecure AS was a risk factor for decreased BMD, regardless of depression. Insecure AS may play a role in the relationship between MDD and BMD or may constitute a risk factor itself. Therapeutic interventions focused on AS could improve psychiatric disorders and physical diseases related to low BMD.

Asunto(s)

Densidad Ósea , Trastorno Depresivo Mayor/psicología , Apego a Objetos , Osteoporosis Posmenopáusica/psicología , Posmenopausia/psicología , Absorciometría de Fotón , Anciano , Trastorno Depresivo Mayor/complicaciones , Trastorno Depresivo Mayor/fisiopatología , Femenino , Humanos , Relaciones Interpersonales , Entrevista Psicológica , Persona de Mediana Edad , Osteoporosis Posmenopáusica/complicaciones , Osteoporosis Posmenopáusica/fisiopatología , Pacientes Ambulatorios/psicología , Inventario de Personalidad , Proyectos Piloto , Posmenopausia/fisiología , Factores de Riesgo , Apoyo Social , Encuestas y Cuestionarios

RESUMEN

Cells are constantly exposed to endogenous and exogenous factors that threaten the integrity of their DNA. The maintenance of genome stability is of paramount importance in the prevention of both cancer and aging processes. To deal with DNA damage, cells put into operation a sophisticated and coordinated mechanism, collectively known as DNA damage response (DDR). The DDR orchestrates different cellular processes, such as DNA repair, senescence and apoptosis. Among the key factors of the DDR, the related proteins p53, p63 and p73, all belonging to the same family of transcription factors, play multiple relevant roles. Indeed, the members of this family are directly involved in the induction of cell cycle arrest that is necessary to allow the cells to repair. Alternatively, they can promote cell death in case of prolonged or irreparable DNA damage. They also take part in a more direct task by modulating the expression of core factors involved in the process of DNA repair or by directly interacting with them. In this review we will analyze the fundamental roles of the p53 family in the aging process through their multifaceted function in DDR.

Asunto(s)

Envejecimiento/fisiología , Reparación del ADN/fisiología , Mamíferos/metabolismo , Proteína p53 Supresora de Tumor/metabolismo , Animales , Regulación de la Expresión Génica , Proteína p53 Supresora de Tumor/genética