RESUMO
Ocular adnexal extranodal marginal zone lymphoma (OA-EMZL) is the most frequent subtype of ocular adnexal lymphoma, with a high propensity for recurrence. Distant recurrence (DR) as an essential prognostic event has unique clinical risk factors, but whether distinct molecular features exist remains poorly understood. Here, we identified potential biomarkers using proteomic analysis of 27 OA-EMZL samples. The MYC-targeted genes PCNA, MCM6, and MCM4 were identified as candidates. MYC-targeted genes were further identified as the most significantly activated gene set in patients with DR. The candidate genes were verified in samples from 11 patients with DR and 33 matched controls using immunohistochemistry. The 3-year and 5-year AUC values of MCM6 (0.699 and 0.757) were higher than those of Ki-67 (0.532 and 0.592). High expressions of MCM6 and MCM4 were significantly associated with shorter distant recurrence-free survival (Log-rank p = 0.017, Log-rank p = 0.0053). Multivariate Cox regression identified MCM6 expression as an independent risk factor for DR (HR, 6.86; 95% CI, 1.32-35.79; P = 0.02). Knockdown of c-Myc in B cells resulted in decreased MCM6 and MCM4 expression and reduced proliferative capacity. Our results suggest that activation of the MYC-targeted gene is a distinct molecular feature of DR in OA-EMZL. MYC-targeted gene, MCM6, is a promising pathological biomarker for DR.
Assuntos
Neoplasias Oculares , Linfoma de Zona Marginal Tipo Células B , Humanos , Proteômica , Neoplasias Oculares/genética , Neoplasias Oculares/metabolismo , Linfoma de Zona Marginal Tipo Células B/patologia , Prognóstico , Imuno-HistoquímicaRESUMO
Gluten is a key component that allows wheat flour to form a dough, and it is also a byproduct of the production of wheat starch. As a commercial product, wheat gluten is increasingly used in the food-related industry because of its versatile functional properties and wide range of sources. Wheat gluten is manufactured industrially on a large scale through the Martin process and batter process and variants thereof. Gliadin and glutenin impart cohesiveness and elasticity properties, respectively, to wheat gluten. The formation of gluten networks and polymers depends mainly on covalent bonds (disulfide bonds) and noncovalent bonds (ionic bonds, hydrogen bonds, and hydrophobic interactions). The multifunctional properties (viscoelasticity, gelation, foamability, etc.) of wheat gluten are shown by rehydration and other processing techniques. Wheat gluten has been widely used in wheat-based products, food auxiliary agents, food packaging, encapsulation and release of food functional ingredients, food adsorption and heat insulation materials, special purpose foods, and versatile applications. In the future, wheat gluten protein will be used as an important raw material to participate in the development and preparation of various food and degradable materials, and the application potential of wheat gluten in food-related industries will be massive. This review summarizes the main manufacturing processes, composition, and structure of gluten protein, and the various functional properties that support its application in the food and related industries.
The versatile functional properties of wheat gluten are closely related to structureThe application as a binder and substitute for meat is growing in popularityRenewable and degradable gluten-based materials are important exploited fieldsWheat gluten shows vast potential and application value in food-related industry.
Assuntos
Farinha , Triticum , Triticum/química , Farinha/análise , Glutens , Indústria de Processamento de Alimentos , ViscosidadeRESUMO
Chromatin remodeling impacts the structural neighborhoods and regulates gene expression. However, the role of enhancer-guided chromatin remodeling in the gene regulation remains unclear. Here, using RNA-seq and ChIP-seq, we identified for the first time that neurotensin (NTS) serves as a key oncogene in uveal melanoma and that CTCF interacts with the upstream enhancer of NTS and orchestrates an 800 kb chromosomal loop between the promoter and enhancer. Intriguingly, this novel CTCF-guided chromatin loop was ubiquitous in a cohort of tumor patients. In addition, a disruption in this chromosomal interaction prevented the histone acetyltransferase EP300 from embedding in the promoter of NTS and resulted in NTS silencing. Most importantly, in vitro and in vivo experiments showed that the ability of tumor formation was significantly suppressed via deletion of the enhancer by CRISPR-Cas9. These studies delineate a novel onco-enhancer guided epigenetic mechanism and provide a promising therapeutic concept for disease therapy.
Assuntos
Fator de Ligação a CCCTC/genética , Carcinogênese/genética , Proteína p300 Associada a E1A/genética , Melanoma/genética , Neurotensina/genética , Neoplasias Uveais/genética , Animais , Fator de Ligação a CCCTC/metabolismo , Sistemas CRISPR-Cas , Carcinogênese/metabolismo , Carcinogênese/patologia , Estudos de Casos e Controles , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Proteína p300 Associada a E1A/metabolismo , Elementos Facilitadores Genéticos , Epigênese Genética , Regulação Neoplásica da Expressão Gênica , Humanos , Masculino , Melanócitos/metabolismo , Melanócitos/patologia , Melanoma/metabolismo , Melanoma/mortalidade , Melanoma/patologia , Camundongos , Camundongos Nus , Neurotensina/antagonistas & inibidores , Neurotensina/metabolismo , Regiões Promotoras Genéticas , Deleção de Sequência , Análise de Sobrevida , Carga Tumoral , Neoplasias Uveais/metabolismo , Neoplasias Uveais/mortalidade , Neoplasias Uveais/patologia , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Aberrant chromatin transformation dysregulates gene expression and may be an important driver of tumorigenesis. However, the functional role of chromosomal dynamics in tumorigenesis remains to be elucidated. Here, using in vitro and in vivo experiments, we reveal a novel long noncoding (lncing) driver at chr12p13.3, in which a novel lncRNA GALNT8 Antisense Upstream 1 (GAU1) is initially activated by an open chromatin status, triggering recruitment of the transcription elongation factor TCEA1 at the oncogene GALNT8 promoter and cis-activates the expression of GALNT8. Analysis of The Cancer Genome Atlas (TCGA) clinical database revealed that the GAU1/GALNT8 driver serves as an important indicative biomarker, and targeted silencing of GAU1 via the HKP-encapsulated method exhibited therapeutic efficacy in orthotopic xenografts. Our study presents a novel oncogenetic mechanism in which aberrant tuning of the chromatin state at specific chromosomal loci exposes factor-binding sites, leading to recruitment of trans-factor and activation of oncogenetic driver, thereby provide a novel alternative concept of chromatin dynamics in tumorigenesis.
Assuntos
Carcinogênese/genética , Cromossomos Humanos Par 12 , Regulação Neoplásica da Expressão Gênica , N-Acetilgalactosaminiltransferases/genética , RNA Longo não Codificante/metabolismo , Adulto , Animais , Biomarcadores Tumorais , Linhagem Celular , Células Cultivadas , Cromatina/metabolismo , Progressão da Doença , Humanos , Masculino , Camundongos Endogâmicos BALB C , Camundongos Nus , N-Acetilgalactosaminiltransferases/metabolismo , Neuroblastoma/genética , Neuroblastoma/metabolismo , Regiões Promotoras Genéticas , RNA Longo não Codificante/genética , Neoplasias da Retina/genética , Neoplasias da Retina/metabolismo , Neoplasias da Retina/patologia , Retinoblastoma/genética , Retinoblastoma/metabolismo , Retinoblastoma/patologia , Fatores de Elongação da Transcrição/metabolismo , Polipeptídeo N-AcetilgalactosaminiltransferaseRESUMO
BACKGROUND: Dynamic N6-methyladenosine (m6A) RNA modification generated and erased by N6-methyltransferases and demethylases regulates gene expression, alternative splicing and cell fate. Ocular melanoma, comprising uveal melanoma (UM) and conjunctival melanoma (CM), is the most common primary eye tumor in adults and the 2nd most common melanoma. However, the functional role of m6A modification in ocular melanoma remains unclear. METHODS: m6A assays and survival analysis were used to explore decreased global m6A levels, indicating a late stage of ocular melanoma and a poor prognosis. Multiomic analysis of miCLIP-seq, RNA-seq and Label-free MS data revealed that m6A RNA modification posttranscriptionally promoted HINT2 expression. RNA immunoprecipitation (RIP)-qPCR and dual luciferase assays revealed that HINT2 mRNA specifically interacted with YTHDF1. Furthermore, polysome profiling analysis indicated a greater amount of HINT2 mRNA in the translation pool in ocular melanoma cells with higher m6A methylation. RESULTS: Here, we show that RNA methylation significantly inhibits the progression of UM and CM. Ocular melanoma samples showed decreased m6A levels, indicating a poor prognosis. Changes in global m6A modification were highly associated with tumor progression in vitro and in vivo. Mechanistically, YTHDF1 promoted the translation of methylated HINT2 mRNA, a tumor suppressor in ocular melanoma. CONCLUSIONS: Our work uncovers a critical function for m6A methylation in ocular melanoma and provides additional insight into the understanding of m6A modification.
Assuntos
Adenosina/análogos & derivados , Neoplasias Oculares/genética , Melanoma/genética , Proteínas Mitocondriais/genética , Processamento Pós-Transcricional do RNA , RNA Mensageiro/genética , Adenosina/metabolismo , Apoptose , Linhagem Celular Tumoral , Transformação Celular Neoplásica/genética , Transformação Celular Neoplásica/metabolismo , Neoplasias Oculares/metabolismo , Neoplasias Oculares/mortalidade , Neoplasias Oculares/patologia , Perfilação da Expressão Gênica , Humanos , Melanoma/metabolismo , Melanoma/mortalidade , Melanoma/patologia , Metilação , Proteínas Mitocondriais/metabolismo , Prognóstico , RNA Mensageiro/metabolismoRESUMO
N6-methyladenosine (m6A), the most prevalent modification of mammalian RNA, has received increasing attention. Although m6A has been shown to be associated with biological activities, such as spermatogenesis modulation, cell spermatogenesis and pluripotency, Drosophila sex determination, and the control of T cell homeostasis and response to heat shock, little is known about its roles in cancer biology and cancer stem cells. Recent articles have noted that some genes have abnormal m6A expression after tumorigenesis, including genes ABS2, RARA, MYB, MYC, ADAM19 and FOX1. Abnormal changes in the m6A levels of these genes are closely related to tumour occurrence and development. In this review, we summarized the 'dual edge weapon' role of RNA methylation in the tumorigenesis. We discussed RNA methylation could lead to not only tumour progression but also tumour suppression. Moreover, we clarified that the abnormal changes in the m6A enrichment of specific loci contribute to tumour occurrence and development, thereby representing a novel anti-cancer strategy by restoration to balanced RNA methylation in tumour cells.
Assuntos
Adenosina/análogos & derivados , Carcinogênese/genética , RNA Mensageiro/genética , Adenosina/genética , Progressão da Doença , Redes Reguladoras de Genes , Humanos , Neoplasias/genéticaRESUMO
Exploring gene function is critical for understanding the complexity of life. DNA sequences and the three-dimensional organization of chromatin (chromosomal interactions) are considered enigmatic factors underlying gene function, and interactions between two distant fragments can regulate transactivation activity via mediator proteins. Thus, a series of chromosome conformation capture techniques have been developed, including chromosome conformation capture (3C), circular chromosome conformation capture (4C), chromosome conformation capture carbon copy (5C), and high-resolution chromosome conformation capture (Hi-C). The application of these techniques has expanded to various fields, but cancer remains one of the major topics. Interactions mediated by proteins or long noncoding RNAs (lncRNAs) are typically found using 4C-sequencing and chromatin interaction analysis by paired-end tag sequencing (ChIA-PET). Currently, Hi-C is used to identify chromatin loops between cancer risk-associated single-nucleotide polymorphisms (SNPs) found by genome-wide association studies (GWAS) and their target genes. Chromosomal conformations are responsible for altered gene regulation through several typical mechanisms and contribute to the biological behavior and malignancy of different tumors, particularly prostate cancer, breast cancer and hematologic neoplasms. Moreover, different subtypes may exhibit different 3D-chromosomal conformations. Thus, C-tech can be used to help diagnose cancer subtypes and alleviate cancer progression by destroying specific chromosomal conformations. Here, we review the fundamentals and improvements in chromosome conformation capture techniques and their clinical applications in cancer to provide insight for future research.
Assuntos
Cromatina/genética , Cromossomos Humanos/química , Neoplasias/genética , Cromatina/química , Mapeamento Cromossômico , Regulação Neoplásica da Expressão Gênica , Predisposição Genética para Doença , Humanos , Conformação de Ácido NucleicoRESUMO
Phthalate esters (PAEs) are plasticizers widely used in the industry and easily released into the environment, posing a serious threat to human health. Molecularly imprinted polymers (MIPs) are important as selective adsorbents for the removal of PAEs. In this study, three kinds of mussel-inspired MIPs for the removal of PAEs were first prepared with gallic acid (GA), hexanediamine (HD), tannic acid (TA), and dopamine (DA) under mild conditions. The adsorption results showed that the MIP with low cost derived from GA and HD (GAHD-MIP) obtained the highest adsorption capacity among these materials. Furthermore, 97.43% of equilibrium capacity could be reached within the first 5 min of adsorption. Especially, the dummy template of diallyl phthalate (DAP) with low toxicity was observed to be more suitable to prepare MIPs than dibutyl phthalate (DBP), although DBP was the target of adsorption. The adsorption process was in accordance with the pseudo-second-order kinetics model. In the isotherm analysis, the adsorption behavior agreed with the Freundlich model. Additionally, the material maintained high adsorption performance after 7 cycles of regeneration tests. The GAHD-MIP adsorbents in this study, with low cost, rapid adsorption equilibrium, green raw materials, and low toxicity dummy template, provide a valuable reference for the design and development of new MIPs.
Assuntos
Dibutilftalato , Ácido Gálico , Polímeros Molecularmente Impressos , Poluentes Químicos da Água , Adsorção , Dibutilftalato/química , Poluentes Químicos da Água/química , Ácido Gálico/química , Polímeros Molecularmente Impressos/química , Ácidos Ftálicos/química , Cinética , Purificação da Água/métodosRESUMO
Cancer-associated fibroblasts (CAFs) exhibit notable versatility, plasticity, and robustness, actively participating in cancer progression through intricate interactions within the tumor microenvironment (TME). N6-methyladenosine (m6A) modification is the most prevalent modification in eukaryotic mRNA, playing essential roles in mRNA metabolism and various biological processes. Howbeit, the precise involvement of m6A in CAF activation remains enigmatic. In this study, we revealed that the m6A demethylase FTO supports CAF-mediated angiogenesis through activation of EGR1 and VEGFA in conjunctival melanoma (CoM). First, single-cell transcriptome analysis revealed that FTO was specifically upregulated in the CAF population, thereby contributing to the hypo-m6A status in the TME of CoM. Moreover, CAFs of CoM displayed extensive proangiogenic potential, which was largely compromised by FTO inhibition, both in vitro and in vivo. By employing multi-omics analysis, we showed that FTO effectively eliminates the m6A modifications of VEGFA and EGR1. This process subsequently disrupts the YTHDF2-dependent mRNA decay pathway, resulting in increased mRNA stability and upregulated expression of these molecules. Collectively, our findings initially indicate that the upregulation of FTO plays a pivotal role in tumor development by promoting CAF-mediated angiogenesis. Therapeutically, targeting FTO may show promise as a potential antiangiogenic strategy to optimize cancer treatment.
Assuntos
Adenosina , Dioxigenase FTO Dependente de alfa-Cetoglutarato , Fibroblastos Associados a Câncer , Proteína 1 de Resposta de Crescimento Precoce , Neovascularização Patológica , Microambiente Tumoral , Fator A de Crescimento do Endotélio Vascular , Dioxigenase FTO Dependente de alfa-Cetoglutarato/genética , Dioxigenase FTO Dependente de alfa-Cetoglutarato/metabolismo , Fibroblastos Associados a Câncer/metabolismo , Fibroblastos Associados a Câncer/patologia , Humanos , Neovascularização Patológica/genética , Neovascularização Patológica/metabolismo , Proteína 1 de Resposta de Crescimento Precoce/genética , Proteína 1 de Resposta de Crescimento Precoce/metabolismo , Animais , Fator A de Crescimento do Endotélio Vascular/metabolismo , Fator A de Crescimento do Endotélio Vascular/genética , Adenosina/análogos & derivados , Adenosina/metabolismo , Camundongos , Estabilidade de RNA , Linhagem Celular Tumoral , Regulação Neoplásica da Expressão Gênica , Proteínas de Ligação a RNA/genética , Proteínas de Ligação a RNA/metabolismo , Melanoma/genética , Melanoma/patologia , Melanoma/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Indutores da Angiogênese/metabolismoRESUMO
Extrachromosomal circular DNA (eccDNA), ranging in size from tens to millions of base pairs, is independent of conventional chromosomes. Recently, eccDNAs have been considered an unanticipated major source of somatic rearrangements, contributing to genomic remodeling through chimeric circularization and reintegration of circular DNA into the linear genome. In addition, the origin of eccDNA is considered to be associated with essential chromatin-related events, including the formation of super-enhancers and DNA repair machineries. Moreover, our understanding of the properties and functions of eccDNA has continuously and greatly expanded. Emerging investigations demonstrate that eccDNAs serve as multifunctional molecules in various organisms during diversified biological processes, such as epigenetic remodeling, telomere trimming, and the regulation of canonical signaling pathways. Importantly, its special distribution potentiates eccDNA as a measurable biomarker in many diseases, especially cancers. The loss of eccDNA homeostasis facilitates tumor initiation, malignant progression, and heterogeneous evolution in many cancers. An in-depth understanding of eccDNA provides novel insights for precision cancer treatment. In this review, we summarized the discovery history of eccDNA, discussed the biogenesis, characteristics, and functions of eccDNA. Moreover, we emphasized the role of eccDNA during tumor pathogenesis and malignant evolution. Therapeutically, we summarized potential clinical applications that target aberrant eccDNA in multiple diseases.
Assuntos
DNA Circular , Neoplasias , Cromatina , DNA Circular/genética , Humanos , Neoplasias/genéticaRESUMO
To understand the formation process of dough with different hydration levels upon mixing and the response of dough rheology, the dynamic evolution of gluten protein was tracked and quantified at morphological, structural, and molecular levels. Both macroscopical and microscopic distribution images showed that partial and full hydration induced quick formation of a more compact gluten network compared with limited hydration. Gluten network in highly hydrated samples was more susceptible to the formation and collapse induced by mechanical force. SE-HPLC results indicated significant depolymerization of glutenin macropolymer (GMP) in fully and partially hydrated samples. Sufficient mixing was accompanied by the increase of ionic and hydrogen bonds, while excessive mixing increased exposure of free -SH. Higher hydration level induced more ordered secondary structure. Correlation and principal component analysis revealed the patterns and dynamics of gluten evolution during dough formation with different hydration levels, and their contribution to the changes in dough modulus.
RESUMO
To understand the formation of gluten network and its regulation on noodle qualities upon mixing and resting, the dynamic distribution and molecular transformation of gluten were tracked and quantified. Confocal laser scanning microscopy and scanning electron microscopy images showed that appropriate mixing (8 min) and resting (60 min) induced a compact gluten network with higher gluten junctions. Both height and width of protein molecular chains were increased by hydration during mixing and reduced after excessive resting (90 min). According to the size exclusion/reversed phase-HPLC profiles, mixing induced slight depolymerization of large glutenin polymer, and α-gliadin subunits were more susceptible to polymerization after appropriate mixing and resting. Increased mixing time was accompanied by the strengthening of ionic and hydrogen bonds, and the weakening of hydrophobic interaction. PCA and correlation analysis revealed the accurate regulation of mixing and resting induced dynamic distribution and evolution of gluten on the macroscopic noodle qualities.
Assuntos
Farinha , Triticum , Pão , Gliadina , Glutens/química , Triticum/químicaRESUMO
Uveal melanoma (UM) and retinoblastoma (RB), which cause blindness and even death, are the most frequently observed primary intraocular malignancies in adults and children, respectively. Epigenetic studies have shown that changes in the epigenome contribute to the rapid progression of both UM and RB following classic genetic changes. The loss of epigenetic homeostasis plays an important role in oncogenesis by disrupting the normal patterns of gene expression. The targetable nature of epigenetic modifications provides a unique opportunity to optimize treatment paradigms and establish new therapeutic options for both UM and RB with these aberrant epigenetic modifications. We aimed to review the research findings regarding relevant epigenetic changes in UM and RB. Herein, we 1) summarize the literature, with an emphasis on epigenetic alterations, including DNA methylation, histone modifications, RNA modifications, noncoding RNAs and an abnormal chromosomal architecture; 2) elaborate on the regulatory role of epigenetic modifications in biological processes during tumorigenesis; and 3) propose promising therapeutic candidates for epigenetic targets and update the list of epigenetic drugs for the treatment of UM and RB. In summary, we endeavour to depict the epigenetic landscape of primary intraocular malignancy tumorigenesis and provide potential epigenetic targets in the treatment of these tumours.
Assuntos
Neoplasias da Retina , Retinoblastoma , Adulto , Carcinogênese/genética , Criança , Epigênese Genética , Homeostase/genética , Humanos , Melanoma , Neoplasias da Retina/genética , Retinoblastoma/genética , Neoplasias UveaisRESUMO
N6-methyladenosine (m6A) is the most universal internal RNA modification on messenger RNAs and regulates the fate and functions of m6A-modified transcripts through m6A-specific binding proteins. Nevertheless, the functional role and potential mechanism of the m6A reading proteins in ocular melanoma tumorigenicity, especially cancer stem-like cell (CSC) properties, remain to be elucidated. Herein, we demonstrated that the m6A reading protein YTHDF3 promotes the translation of the target transcript CTNNB1, contributing to ocular melanoma propagation and migration through m6A methylation. YTHDF3 is highly expressed in ocular melanoma stem-like cells and abundantly enriched in ocular melanoma tissues, which is related to poor clinical prognosis. Moreover, YTHDF3 is required for the maintenance of CSC properties and tumor initiation capacity in ocular melanoma both in vitro and in vivo. Ocular melanoma cells with targeted YTHDF3 knockdown exhibited inhibitory tumor proliferation and migration abilities. Transcriptome-wide mapping of m6A peaks and YTHDF3 binding peaks on mRNAs revealed a key target gene candidate, CTNNB1. Mechanistically, YTHDF3 enhances CTNNB1 translation through recognizing and binding the m6A peaks on CTNNB1 mRNA.
Assuntos
RNA MensageiroRESUMO
PURPOSE: All uveal melanoma and a fraction of other melanoma subtypes are driven by activation of the G-protein alpha-q (Gαq) pathway. Targeting these melanomas has proven difficult despite advances in the molecular understanding of key driver signaling pathways in the disease pathogenesis. Inhibitors of Gαq have shown promising preclinical results, but their therapeutic activity in distinct Gαq mutational contexts and in vivo have remained elusive. EXPERIMENTAL DESIGN: We used an isogenic melanocytic cellular system to systematically examine hotspot mutations in GNAQ (e.g., G48V, R183Q, Q209L) and CYSLTR2 (L129Q) found in human uveal melanoma. This cellular system and human uveal melanoma cell lines were used in vitro and in in vivo xenograft studies to assess the efficacy of Gαq inhibition as a single agent and in combination with MEK inhibition. RESULTS: We demonstrate that the Gαq inhibitor YM-254890 inhibited downstream signaling and in vitro growth in all mutants. In vivo, YM-254890 slowed tumor growth but did not cause regression in human uveal melanoma xenografts. Through comprehensive transcriptome analysis, we observed that YM-254890 caused inhibition of the MAPK signaling with evidence of rebound by 24 hours and combination treatment of YM-254890 and a MEK inhibitor led to sustained MAPK inhibition. We further demonstrated that the combination caused synergistic growth inhibition in vitro and tumor shrinkage in vivo. CONCLUSIONS: These data suggest that the combination of Gαq and MEK inhibition provides a promising therapeutic strategy and improved therapeutic window of broadly targeting Gαq in uveal melanoma.See related commentary by Neelature Sriramareddy and Smalley, p. 1217.
Assuntos
Melanoma , Neoplasias Uveais , Linhagem Celular Tumoral , Subunidades alfa de Proteínas de Ligação ao GTP/genética , Subunidades alfa Gq-G11 de Proteínas de Ligação ao GTP/genética , Humanos , Melanoma/tratamento farmacológico , Melanoma/genética , Quinases de Proteína Quinase Ativadas por Mitógeno/metabolismo , Mutação , Neoplasias Uveais/tratamento farmacológico , Neoplasias Uveais/genéticaRESUMO
Sertoli cells are essential for regulating normal spermatogenesis. However, the mechanisms underlying human Sertoli cell development remain largely elusive. Here we examined the function and signaling pathways of BMP6 in regulating human Sertoli cells. RT-PCR, immunocytochemistry and Western blots revealed that BMP6 and its multiple receptors were expressed in human Sertoli cells. CCK-8 and EDU assays showed that BMP6 promoted the proliferation of Sertoli cells. Conversely, BMP6 siRNAs inhibited the division of these cells. Annexin V/PI assay indicated that BMP6 reduced the apoptosis in human Sertoli cells, whereas BMP6 knockdown assumed reverse effects. BMP6 enhanced the expression levels of ZO1, SCF, GDNF and AR in human Sertoli cells, and ELISA assay showed an increase of SCF by BMP6 and a reduction by BMP6 siRNAs. Notably, Smad2/3 phosphorylation and cyclin D1 were enhanced by BMP6 and decreased by BMP6 siRNAs in human Sertoli cells. The levels of DACH1 and TFAP2A were increased by BMP6 and reduced by BMP6 siRNAs, and the growth of human Sertoli cells was inhibited by these siRNAs. Collectively, these results suggest that BMP6 regulates the proliferation and apoptosis of human Sertoli cells via activating the Smad2/3/cyclin D1 and DACH1 and TFAP2A pathway.
Assuntos
Proteína Morfogenética Óssea 6/metabolismo , Ciclina D1/metabolismo , Proteínas do Olho/genética , Células de Sertoli/citologia , Proteínas Smad/metabolismo , Fator de Transcrição AP-2/genética , Fatores de Transcrição/genética , Apoptose , Proteína Morfogenética Óssea 6/genética , Proliferação de Células , Células Cultivadas , Proteínas do Olho/metabolismo , Humanos , Masculino , Fosforilação , Células de Sertoli/metabolismo , Transdução de Sinais , Proteína Smad2/metabolismo , Proteína Smad3/metabolismo , Fator de Transcrição AP-2/metabolismo , Fatores de Transcrição/metabolismo , Ativação TranscricionalRESUMO
Stem cells, by definition, are the primitive cells that have the potential of both self-renewal and differentiation into a number of mature and functional cells, and thus they have great applications in cell therapy and tissue engineering for regenerative medicine. Bone morphogenetic protein 6 (BMP6) belongs to transforming growth factor ß(TGF-ß) superfamily. The fate determinations of stem cells require complex regulatory networks that involve BMP6 signaling pathway. Recent studies have demonstrated that BMP6 plays crucial roles in controlling the self-renewal and differentiation of stem cells. In this review, we address the expression, function and regulation of BMP6 in various kinds of stem cells, with focus on mesenchymal stem cells (MSCs), germline stem cells (GSCs), hematopoietic stem cells (HSCs), and neural stem cells (NSCs). Notably, there are distinct effects of BMP6 on promoting self-renewal and differentiation of these stem cells. We also discuss the involement of BMP6 in diseases, including leukemia, astrocytic glioma, and Alzheimer's disease, and the therapy of these diseases via gene targeting. We further highlight certain issues for further investigation on the regulation and function of BMP6 in stem cells. Significantly, a thorough understanding of BMP6 regulation on a variety of adult stem cells could make them feasible for applications in both regenerative and reproductive medicine, and it would shed novel insights into the etiology of the diseases and offer new targets for drug design to treat these disorders.