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1.
Int J Med Sci ; 16(12): 1557-1563, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31839743

RESUMO

E2F3, a member of the E2F family, plays a critical role in cell cycle and proliferation by targeting downstream, retinoblastoma (RB) a tumor suppressor family protein. The purpose of this study, was to investigate the role and function of E2F3 in vivo. We examined phenotypic abnormalities, by deletion of the E2f3 gene in mice. Complete ablation of the E2F3 was fully penetrant, in the pure C57BL/6N background. The E2f3+/ - mouse embryo developed normally without fatal disorder. However, they exhibited reduced body weight, growth retardation, skeletal imperfection, and poor grip strength ability. Findings suggest that E2F3 has a pivotal role in muscle and bone development, and affect normal mouse growth.


Assuntos
Desenvolvimento Ósseo/genética , Fator de Transcrição E2F3/genética , Desenvolvimento Embrionário/genética , Músculo Esquelético/crescimento & desenvolvimento , Animais , Apoptose/genética , Peso Corporal/genética , Ciclo Celular/genética , Proliferação de Células/genética , Embrião de Mamíferos , Humanos , Camundongos , Camundongos Knockout , Músculo Esquelético/metabolismo , Músculo Esquelético/fisiopatologia , Fenótipo
2.
Stroke ; 50(10): 2651-2660, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31500558

RESUMO

Background and Purpose- Genome-wide association studies have identified the HDAC9 (histone deacetylase 9) gene region as a major risk locus for atherosclerotic stroke and coronary artery disease in humans. Previous results suggest a role of altered HDAC9 expression levels as the underlying disease mechanism. rs2107595, the lead single nucleotide polymorphism for stroke and coronary artery disease resides in noncoding DNA and colocalizes with histone modification marks suggestive of enhancer elements. Methods- To determine the mechanisms by which genetic variation at rs2107595 regulates HDAC9 expression and thus vascular risk we employed targeted resequencing, proteome-wide search for allele-specific nuclear binding partners, chromatin immunoprecipitation, genome-editing, reporter assays, circularized chromosome conformation capture, and gain- and loss-of-function experiments in cultured human cell lines and primary immune cells. Results- Targeted resequencing of the HDAC9 locus in patients with atherosclerotic stroke and controls supported candidacy of rs2107595 as the causative single nucleotide polymorphism. A proteomic search for nuclear binding partners revealed preferential binding of the E2F3/TFDP1/Rb1 complex (E2F transcription factor 3/transcription factor Dp-1/Retinoblastoma 1) to the rs2107595 common allele, consistent with the disruption of an E2F3 consensus site by the risk allele. Gain- and loss-of-function studies showed a regulatory effect of E2F/Rb proteins on HDAC9 expression. Compared with the common allele, the rs2107595 risk allele exhibited higher transcriptional capacity in luciferase assays and was associated with higher HDAC9 mRNA levels in primary macrophages and genome-edited Jurkat cells. Circularized chromosome conformation capture revealed a genomic interaction of the rs2107595 region with the HDAC9 promoter, which was stronger for the common allele as was the in vivo interaction with E2F3 and Rb1 determined by chromatin immunoprecipitation. Gain-of-function experiments in isogenic Jurkat cells demonstrated a key role of E2F3 in mediating rs2107595-dependent transcriptional regulation of HDAC9. Conclusions- Collectively, our findings imply allele-specific transcriptional regulation of HDAC9 via E2F3 and Rb1 as a major mechanism mediating vascular risk at rs2107595.


Assuntos
Aterosclerose/genética , Fator de Transcrição E2F3/genética , Regulação da Expressão Gênica/genética , Histona Desacetilases/genética , Proteínas Repressoras/genética , Proteínas de Ligação a Retinoblastoma/genética , Ubiquitina-Proteína Ligases/genética , Células Cultivadas , Predisposição Genética para Doença/genética , Humanos , Polimorfismo de Nucleotídeo Único
3.
Mol Cell ; 74(6): 1264-1277.e7, 2019 06 20.
Artigo em Inglês | MEDLINE | ID: mdl-31130363

RESUMO

E2F1, E2F2, and E2F3A, the three activators of the E2F family of transcription factors, are key regulators of the G1/S transition, promoting transcription of hundreds of genes critical for cell-cycle progression. We found that during late S and in G2, the degradation of all three activator E2Fs is controlled by cyclin F, the substrate receptor of 1 of 69 human SCF ubiquitin ligase complexes. E2F1, E2F2, and E2F3A interact with the cyclin box of cyclin F via their conserved N-terminal cyclin binding motifs. In the short term, E2F mutants unable to bind cyclin F remain stable throughout the cell cycle, induce unscheduled transcription in G2 and mitosis, and promote faster entry into the next S phase. However, in the long term, they impair cell fitness. We propose that by restricting E2F activity to the S phase, cyclin F controls one of the main and most critical transcriptional engines of the cell cycle.


Assuntos
Ciclo Celular/genética , Ciclinas/genética , Fator de Transcrição E2F1/genética , Fator de Transcrição E2F2/genética , Fator de Transcrição E2F3/genética , Proteínas Ligases SKP Culina F-Box/genética , Transcrição Genética , Linhagem Celular Tumoral , Ciclinas/metabolismo , Fator de Transcrição E2F1/metabolismo , Fator de Transcrição E2F2/metabolismo , Fator de Transcrição E2F3/metabolismo , Células Epiteliais/citologia , Células Epiteliais/metabolismo , Regulação da Expressão Gênica , Aptidão Genética , Células HEK293 , Células HeLa , Humanos , Mutação , Osteoblastos/citologia , Osteoblastos/metabolismo , Proteólise , Proteínas Ligases SKP Culina F-Box/metabolismo , Transdução de Sinais , Ubiquitinação
4.
Mol Med Rep ; 19(6): 4946-4954, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30957179

RESUMO

The potential role of microRNA (miR)­210­3p in carcinogenesis and the cisplatin sensitivity of ovarian cancer were evaluated in the present study. The relative expression levels of miR­210­3p in cisplatin­sensitive SKOV­3 cells and cisplatin­resistant SKOV­3/DDP cells were determined using reverse transcription­quantitative polymerase chain reaction analysis. miR­210­3p mimics and inhibitors were transfected into SKOV­3/DDP cells. Cell Counting Kit­8, scratch and Transwell invasion assays and flow cytometry were conducted to evaluate the role of miR­210­3p in ovarian cancer cells. A luciferase reporter assay was used to verify the association between miR­210­3p and E2F transcription factor 3 (E2F3). Drug sensitivity was evaluated by treating the cells with cisplatin. The expression level of miR­210­3p was lower in SKOV­3/DDP cells than in SKOV­3 cells. Compared with the untransfected control, SKOV­3 cells transfected with miR­210­3p exhibited a significantly higher survival rate. The overexpression of miR­210­3p inhibited SKOV­3/DDP cell proliferation, migration and invasion, and promoted cell apoptosis. By contrast, the inhibition of miR­210­3p promoted cell migration and invasion. The luciferase reporter assay confirmed that E2F3 was a direct target gene of miR­210­3p. Cisplatin treatment resulted in a sharp decrease in the survival rate of SKOV­3/DDP cells transfected with the miR­210­3p mimics. The decrease in cell survival rate caused by the overexpression of miR­210­3p was rescued by the overexpression of E2F3 in SKOV­3/DDP cells. Taken together, these results suggest that miR­210­3p may act as a tumor suppressor in ovarian cancer cells and affect the sensitivity of cells to cisplatin by directly targeting E2F3. This indicates its potential use as a therapeutic target for improving drug resistance in ovarian cancer.


Assuntos
Proliferação de Células/efeitos dos fármacos , Cisplatino/farmacologia , Fator de Transcrição E2F3/metabolismo , MicroRNAs/metabolismo , Regiões 3' não Traduzidas , Antagomirs/metabolismo , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Sequência de Bases , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Resistencia a Medicamentos Antineoplásicos , Fator de Transcrição E2F3/química , Fator de Transcrição E2F3/genética , Feminino , Humanos , MicroRNAs/antagonistas & inibidores , MicroRNAs/genética , Neoplasias Ovarianas/patologia , Alinhamento de Sequência
5.
Chem Biol Interact ; 306: 39-46, 2019 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-30926320

RESUMO

MicroRNAs (miRNAs) have been regarded as potential modulators in varying ocular diseases, including age-related cataract (ARC). However, the roles of miR-221 in ARC progression and its underlying mechanism remain poorly understood. In this study, human lens epithelial cell line (SRA01/04) was used to investigate the potential function of miR-221 in vitro. The expressions of miR-221, sirtuin-1 (SIRT1) and E2F transcription factor 3 (E2F3) were measured in ARC tissues by quantitative real-time polymerase chain reaction and western blotting, respectively. To investigate the effect of miR-221, SIRT1 and E2F3 on cell apoptosis, SRA01/04 cells were transfected with miR-221 inhibitor, negative control inhibitor, pcDNA3.1-SIRT1 overexpression vector, pcDNA3.1-E2F3 overexpression vector or pcDNA3.1 empty vector. After the transfection, cell viability was detected in SRA01/04 cells at 0, 24, 48 or 72 h by cell counting kit-8 assay. Cell apoptosis was evaluated in transfected SRA01/04 cells by flow cytometry and western blotting at 72 h. The interaction between miR-221 and SIRT1 or E2F3 was probed by luciferase activity and RNA immunoprecipitation assays. Results showed that high expression of miR-221 was exhibited in ARC tissues compared with that in normal samples and associated with Lens Opacities Classification System III grades. Knockdown of miR-221 promoted cell viability and inhibited apoptosis in SRA01/04 cells. Moreover, both of SIRT1 and E2F3 levels were directly targeted by miR-221 and down-regulated in ARC tissues. Besides, overexpression of SIRT1 or E2F3 increased cell viability and suppressed apoptosis in SRA01/04 cells, which was reversed by addition of miR-221. We concluded that miR-221 promoted lens epithelial cells apoptosis through regulating SIRT1 and E2F3, providing a novel biomarker for treatment of ARC.


Assuntos
Apoptose , Fator de Transcrição E2F3/metabolismo , Células Epiteliais/metabolismo , Cristalino/metabolismo , MicroRNAs/metabolismo , Sirtuína 1/metabolismo , Sobrevivência Celular , Células Cultivadas , Fator de Transcrição E2F3/genética , Células Epiteliais/patologia , Feminino , Humanos , Cristalino/patologia , Masculino , MicroRNAs/genética , Pessoa de Meia-Idade , Sirtuína 1/genética
6.
Mol Med Rep ; 19(5): 3575-3583, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30864742

RESUMO

MicroRNAs (miRNA/miRs) have been demonstrated to be critical post­transcriptional modulators of gene expression during tumorigenesis. Numerous miRNAs have been revealed to be downregulated in human epithelial ovarian cancer (EOC). In the present study, it was observed that the expression of miR­145 was decreased in EOC tissues and cell lines. Overexpression of miR­145 inhibited the proliferation, migration and invasion of EOC cells. The D­type cyclin 2, cyclin D2 (CCND2), and E2F transcription factor 3 (E2F3) were confirmed to be targets of miR­145. In addition, restoration of these 2 genes significantly reversed the tumor suppressive effects of miR­145. Collectively, the results indicated that miR­145 serves a critical role in suppressing the biological behavior of EOC cells by targeting CCND2 and E2F3. Therefore, miR­145 was suggested to be a potential miRNA­based therapeutic target in ovarian cancer.


Assuntos
Ciclina D2/genética , Fator de Transcrição E2F3/genética , Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , Neoplasias Ovarianas/genética , Neoplasias Ovarianas/patologia , Interferência de RNA , Regiões 3' não Traduzidas , Adulto , Idoso , Ciclo Celular/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células , Feminino , Genes Reporter , Humanos , Pessoa de Meia-Idade
7.
Commun Biol ; 2: 3, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30740539

RESUMO

Mitochondrial damage is caused by changes in the micro-environmental conditions during tumor progression. Cancer cells require mechanisms for mitochondrial quality control during this process; however, how mitochondrial integrity is maintained is unclear. Here we show that E2F3d, a previously unidentified E2F3 isoform, mediates hypoxia-induced mitophagy in cancer cells. Aberrant activity and expression of the E2F3 transcription factor is frequently observed in many cancer cells. Loss of retinoblastoma (Rb) protein family function increases the expression of E2F3d and E2F3a. E2F3d localizes to the outer mitochondrial membrane and its cytosolic domain contains an LC3-interacting region motif. Overexpression of E2F3d induces mitochondrial fragmentation and mitophagy, suggesting that E2F3d plays an important role in mitophagy. Furthermore, depletion of E2F3s attenuates hypoxia-induced mitophagy and increases intracellular levels of reactive oxygen species, which is reversed by the reintroduction of E2F3d. This study presents another key player that regulates mitochondrial quality control in cancer cells.


Assuntos
Hipóxia Celular , Fator de Transcrição E2F3/metabolismo , Mitocôndrias/metabolismo , Proteínas Mitocondriais/metabolismo , Mitofagia , Neoplasias/metabolismo , Dinaminas/genética , Fator de Transcrição E2F3/genética , Técnicas de Silenciamento de Genes , Células HEK293 , Células HeLa , Humanos , Proteínas Associadas aos Microtúbulos/metabolismo , Membranas Mitocondriais/metabolismo , Isoformas de Proteínas/metabolismo , Espécies Reativas de Oxigênio/metabolismo
8.
J BUON ; 23(5): 1492-1499, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30570877

RESUMO

PURPOSE: MicroRNA (miR)-194-5p is downregulated in bladder cancer (BC), but its role in BC has not been determined mechanistically. METHODS: The expression levels of miR-194-5p and E2F transcription factor 3 (E2F3) were determined by means of quantitative reverse transcription and polymerase chain reaction in BC specimens. In addition, T24 BC cells were transfected with a miR-194-5p mimic, a miR-194-5p inhibitor, or E2F3 small interfering (si)RNA, and the level of E2F3 protein expressed by these cells was assessed by western blotting. A dual luciferase reporter assay was applied to verify the binding site between miR-194-5p and the 3' untranslated region of E2F3. Transwell assays were performed to examine cell migration and invasion. RESULTS: Dysregulation of miR-194-5p in BC was closely associated with node metastasis and differentiation. In BC specimens and cell lines, miR-194-5p mRNA was downregulated, while E2F3 mRNA was upregulated. Overexpression of miR-194-5p suppressed the expression of E2F3 mRNA and protein. By regulating E2F3, miR-194-5p inhibited cell migration and invasion in BC. Treatment of BC cells with E2F3 siRNA had the same effect as did overexpression of miR-194-5p. CONCLUSIONS: MiR-194-5p directly targets E2F3 and inhibits cell migration and invasion in BC.


Assuntos
Fator de Transcrição E2F3/metabolismo , MicroRNAs/metabolismo , Neoplasias da Bexiga Urinária/genética , Neoplasias da Bexiga Urinária/metabolismo , Movimento Celular/fisiologia , Regulação para Baixo , Fator de Transcrição E2F3/antagonistas & inibidores , Fator de Transcrição E2F3/biossíntese , Fator de Transcrição E2F3/genética , Humanos , MicroRNAs/biossíntese , MicroRNAs/genética , Invasividade Neoplásica , Transfecção , Neoplasias da Bexiga Urinária/patologia
9.
Eur Rev Med Pharmacol Sci ; 22(24): 8640-8648, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30575904

RESUMO

OBJECTIVE: The aim of this study was to explore the role of microRNA-210 (miR-210) and E2F3 in the development of pancreatic cancer and to investigate the possible underlying mechanism. PATIENTS AND METHODS: The expression level of miR-210 in pancreatic cancer tissues, para-cancerous tissues, and normal pancreatic tissues was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The correlation between miR-210 expression and pathological indicators of pancreatic cancer was analyzed. Meanwhile, the expression of miR-210 in pancreatic cancer cells and normal pancreatic ductal epithelial cells was detected by qRT-PCR. After transfection with miR-210 mimics and inhibitor, the viability and cell cycle of pancreatic cancer cells were detected by cell counting kit-8 (CCK-8) assay and flow cytometry, respectively. The binding condition of miR-210 and E2F3 was verified by Dual-Luciferase reporter gene assay. RESULTS: MiR-210 was lowly expressed in pancreatic cancer tissues than that of para-cancerous tissues. The expression of miR-210 was negatively correlated with TNM stage and tumor size of pancreatic cancer. In vitro experiments showed that the miR-210 was downregulated in pancreatic cancer cells than that of normal pancreatic ductal epithelial cells. Meanwhile, overexpression of miR-210 arrested cell cycle decreased cell viability and downregulated E2F3 expression in pancreatic cancer cells. Dual-Luciferase reporter gene assay indicated that E2F3 bound to mi-210. Further experiments confirmed that E2F3 was negatively regulated by miR-210. CONCLUSIONS: MiR-210 knockdown promotes cell proliferation by upregulating E2F3 expression, thereby promoting the progression of pancreatic cancer.


Assuntos
Fator de Transcrição E2F3/genética , Regulação Neoplásica da Expressão Gênica , MicroRNAs/metabolismo , Neoplasias Pancreáticas/genética , Ciclo Celular/genética , Linhagem Celular Tumoral , Proliferação de Células/genética , Sobrevivência Celular/genética , Regulação para Baixo , Feminino , Técnicas de Silenciamento de Genes , Humanos , Masculino , MicroRNAs/genética , Pessoa de Meia-Idade , Pâncreas/patologia , Neoplasias Pancreáticas/patologia
10.
Biosci Rep ; 38(6)2018 12 21.
Artigo em Inglês | MEDLINE | ID: mdl-30487158

RESUMO

Gastric cancer (GC) is the second most frequent cause of cancer-related mortality in the world, with Eastern Asia having the highest incidence rates. E2F is a family of transcription factor proteins that has a variety of functions, which include control of cell cycle, cell differentiation, DNA damage response and cell death. E2F transcription factors are divided into two subfamilies: transcription activators (E2F transcription factors 1 (E2F1), 2 (E2F2) and 3a (E2F3a)) and repressors (E2F3b, E2F transcription factors 4 (E2F4), 5 (E2F5), 6 (E2F6), 7 (E2F7) and 8 (E2F8)). Studies have demonstrated that E2F had prognostic significance in a number of cancers. However, the entirety of the prognostic roles of E2F mRNA expression in GC has not yet been apparently determined. In the present study, the prognostic value of individual family members of E2F mRNA expression for overall survival (OS) was evaluated by using online Kaplan-Meier Plotter (KM Plotter) database. Our result demonstrated that high expressions of three family members of E2F (E2F1, E2F3, E2F4) mRNA were significantly associated with unfavourable OS in all GC patients. However, increased expressions of E2F2, E2F5, E2F6 and E2F7 were significantly associated with favourable OS, especially for higher clinical stages in GC patients. These results provided a better insight into the prognostic functions of E2F mRNA genes in GC. Although the results should be further verified in clinical trials, our findings may be a favourable prognostic predictor for the development of newer therapeutic drugs in the treatment of GC.


Assuntos
Proteínas de Ligação a DNA/genética , Fatores de Transcrição E2F/genética , Prognóstico , Neoplasias Gástricas/genética , Divisão Celular/genética , Intervalo Livre de Doença , Fator de Transcrição E2F1/genética , Fator de Transcrição E2F2/genética , Fator de Transcrição E2F3/genética , Feminino , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Estimativa de Kaplan-Meier , Masculino , Valor Preditivo dos Testes , RNA Mensageiro/genética , Neoplasias Gástricas/epidemiologia , Neoplasias Gástricas/patologia
11.
Cell Death Dis ; 9(5): 509, 2018 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-29724991

RESUMO

HOXB9, as a HOX family transcription factor, playing a significant role in embryonic development and cancer progression. However, the function of HOXB9 and its precise mechanism in regulating endometrial cancer progression remains unknown. Here, we demonstrated that the expression of HOXB9 was increased in endometrial cancer, and associated with histological grade and lymph node metastasis. In addition, elevated HOXB9 predicts a poor prognosis in endometrial cancer patients. Interestingly, bioinformatics analysis of TCGA cancer database showed that HOXB9 expression is positively correlated with E2F3 expression. Moreover, HOXB9 promoted E2F3 expression by directly targeting to its promoter. Furthermore, we found that knocking down E2F3 abolished the ability of HOXB9 in enhancing cell migration. Taken together, for the first, we demonstrated the function and mechanism of HOXB9 in regulating endometrial cancer progression, and indicated HOXB9 may be a novel prognostic marker of endometrial cancer.


Assuntos
Adenocarcinoma/genética , Fator de Transcrição E2F3/genética , Neoplasias do Endométrio/genética , Regulação Neoplásica da Expressão Gênica , Proteínas de Homeodomínio/genética , Adenocarcinoma/metabolismo , Adenocarcinoma/mortalidade , Adenocarcinoma/patologia , Adulto , Idoso , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Progressão da Doença , Fator de Transcrição E2F3/antagonistas & inibidores , Fator de Transcrição E2F3/metabolismo , Neoplasias do Endométrio/metabolismo , Neoplasias do Endométrio/mortalidade , Neoplasias do Endométrio/patologia , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Feminino , Proteínas de Homeodomínio/antagonistas & inibidores , Proteínas de Homeodomínio/metabolismo , Humanos , Metástase Linfática , Pessoa de Meia-Idade , Gradação de Tumores , Estadiamento de Neoplasias , Prognóstico , Regiões Promotoras Genéticas , Ligação Proteica , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Transdução de Sinais , Análise de Sobrevida
12.
Mol Med Rep ; 18(1): 1155-1164, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29767254

RESUMO

MicroRNAs (miRNA/miRs) have been associated with the initiation and progression of non­small­cell lung cancer (NSCLC). Hence, a comprehensive understanding of the association between dysregulated miRNAs and NSCLC may contribute to the identification of novel therapeutic methods for patients with NSCLC. MiRNA­433 (miR­433) has been reported to be dysregulated in numerous types of human cancers; however, its expression pattern, biological roles and associated mechanisms in NSCLC require further investigation. The present study aimed to detect miR­433 expression and determine its roles and underlying molecular mechanisms in NSCLC. In the present study, reverse transcription­quantitative polymerase chain reaction revealed that miR­433 was significantly downregulated in NSCLC tissues and cell lines. This decreased miR­433 expression was strongly associated with the tumor node metastasis stage and lymph node metastasis of patients with NSCLC. Cell Counting kit­8 and cell invasion assays revealed that the resumption of miR­433 expression decreased the proliferation and invasion of NSCLC cells. Bioinformatics analysis predicted E2F transcription factor 3 (E2F3) as a potential target of miR­433. Luciferase reporter assay, RT­qPCR and western blot analysis further demonstrated that E2F3 was a direct target of miR­433 in NSCLC. E2F3 downregulation induced by small interfering RNA exhibited inhibitory effects similar to those of miR­433 overexpression in NSCLC cells, and the restored E2F3 expression counteracted the suppressive effects on NSCLC cells induced by miR­433 overexpression. Therefore, miR­433 may inhibit the progression of NSCLC, at least in part, by targeting E2F3. The present study indicated that miR­433 may be investigated as an innovative candidate target for the therapy of patients with this fatal disease.


Assuntos
Carcinoma Pulmonar de Células não Pequenas/metabolismo , Proliferação de Células , Fator de Transcrição E2F3/metabolismo , Neoplasias Pulmonares/metabolismo , MicroRNAs/biossíntese , Proteínas de Neoplasias/metabolismo , RNA Neoplásico/biossíntese , Células A549 , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/patologia , Fator de Transcrição E2F3/genética , Feminino , Humanos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , Masculino , MicroRNAs/genética , Invasividade Neoplásica , Proteínas de Neoplasias/genética , RNA Neoplásico/genética
13.
Biochem Biophys Res Commun ; 502(3): 358-363, 2018 07 20.
Artigo em Inglês | MEDLINE | ID: mdl-29807010

RESUMO

As the development of sequencing technology, more and more circular RNAs (circRNAs) are identified in human cancer tissues. Increasing evidences imply circRNAs are important regulators in tumor progression. Nevertheless, how circRNAs participate in breast cancer development and progression is not well understood. In the present study, we identified a novel circRNA hsa_circ_0008039 with upregulated expression level in breast cancer tissues. By functional experiments, we found that hsa_circ_0008039 depletion significantly suppressed the proliferation, arrested cell-cycle progression and reduced migration in breast cancer. Mechanistic investigations suggested that hsa_circ_0008039 served as a competing endogenous RNA (ceRNA) of miR-432-5p. Subsequently, E2F3 was identified as the functional target of miR-432-5p and overexpression of hsa_circ_0008039 elevated E2F3 expression in breast cancer. On the whole, our study indicated that hsa_circ_0008039 exerted oncogenic roles in breast cancer and suggested the hsa_circ_0008039/miR-432-5p/E2F3 axis might be a potential therapeutic target.


Assuntos
Neoplasias da Mama/genética , Fator de Transcrição E2F3/genética , MicroRNAs/genética , RNA/genética , Animais , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Fator de Transcrição E2F3/metabolismo , Feminino , Regulação Neoplásica da Expressão Gênica , Técnicas de Silenciamento de Genes , Xenoenxertos , Humanos , Células MCF-7 , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , MicroRNAs/metabolismo , Oncogenes , RNA/antagonistas & inibidores , RNA/metabolismo , RNA Circular , Regulação para Cima
14.
PLoS One ; 13(4): e0194937, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29617434

RESUMO

The E2F transcription factors control key elements of development, including mammary gland branching morphogenesis, with several E2Fs playing essential roles. Additional prior data has demonstrated that loss of individual E2Fs can be compensated by other E2F family members, but this has not been tested in a mammary gland developmental context. Here we have explored the role of the E2Fs and their ability to functionally compensate for each other during mammary gland development. Using gene expression from terminal end buds and chromatin immunoprecipitation data for E2F1, E2F2 and E2F3, we noted both overlapping and unique mammary development genes regulated by each of the E2Fs. Based on our computational findings and the fact that E2Fs share a common binding motif, we hypothesized that E2F transcription factors would compensate for each other during mammary development and function. To test this hypothesis, we generated RNA from E2F1-/-, E2F2-/- and E2F3+/- mouse mammary glands. QRT-PCR on mammary glands during pregnancy demonstrated increases in E2F2 and E2F3a in the E2F1-/- mice and an increase in E2F2 levels in E2F3+/- mice. During lactation we noted that E2F3b transcript levels were increased in the E2F2-/- mice. Given that E2Fs have previously been noted to have the most striking effects on development during puberty, we hypothesized that loss of individual E2Fs would be compensated for at that time. Double mutant mice were generated and compared with the single knockouts. Loss of both E2F1 and E2F2 revealed a more striking phenotype than either knockout alone, indicating that E2F2 was compensating for E2F1 loss. Interestingly, while E2F2 was not able to functionally compensate for E2F3+/- during mammary outgrowth, increased E2F2 expression was observed in E2F3+/- mammary glands during pregnancy day 14.5 and lactation day 5. Together, these findings illustrate the specificity of E2F family members to compensate during development of the mammary gland.


Assuntos
Fator de Transcrição E2F1/metabolismo , Fator de Transcrição E2F2/metabolismo , Fator de Transcrição E2F3/metabolismo , Glândulas Mamárias Animais/crescimento & desenvolvimento , Animais , Fator de Transcrição E2F1/deficiência , Fator de Transcrição E2F1/genética , Fator de Transcrição E2F2/deficiência , Fator de Transcrição E2F2/genética , Fator de Transcrição E2F3/deficiência , Fator de Transcrição E2F3/genética , Feminino , Regulação da Expressão Gênica , Células HCT116 , Humanos , Camundongos , Camundongos Knockout , Gravidez , Interferência de RNA , RNA Interferente Pequeno/metabolismo
15.
Cell Death Dis ; 9(3): 370, 2018 03 06.
Artigo em Inglês | MEDLINE | ID: mdl-29511172

RESUMO

Retinoblastoma tumor suppressor (Rb) promotes cell cycle exit, survival, differentiation, and tumor suppression in the retina. Here, we show it is also essential for vascularization and lamination. Despite minimal effects on Hif1a target expression, intraretinal vascular plexi did not form in the Rb -/- murine retina. Deleting adenovirus E2 promoter binding factor 3 (E2f3), which rescues starburst amacrine cell differentiation, or E2f2, had no effect, but deleting E2f1, which promotes neuronal cell cycle exit and survival, restored retinal vasculature. We specifically linked cell loss to the defect because removing Bax rescued rod and bipolar neurons and the vasculature, but not cell cycle exit. Despite rescuing Rb -/- neurons, Bax deletion exacerbated a delay in outer retina lamination, and exposed a requirement for Rb in inner retina lamination. The latter resembled Sem5 or FAT atypical cadherin 3 (Fat3) mutants, but expression of Sem5/Fat3 pathway components, or that of Neogenin, which perturbs migration in the Rb -/- cortex, was unchanged. Instead, lamination defects correlated with ectopic division, and were E2f1-dependent, implicating the cell cycle machinery. These in vivo studies expose new developmental roles for Rb, pinpoint aberrant E2f1 and Bax activity in neuronal death and vascular loss, and further implicate E2f1 in defective lamination. Links between Rb, angiogenesis and lamination have implications for the treatment of neovascularization, neurodegeneration and cancer.


Assuntos
Neovascularização Fisiológica , Retina/crescimento & desenvolvimento , Retina/metabolismo , Vasos Retinianos/metabolismo , Proteína do Retinoblastoma/metabolismo , Animais , Apoptose , Ciclo Celular , Fator de Transcrição E2F2/genética , Fator de Transcrição E2F2/metabolismo , Fator de Transcrição E2F3/genética , Fator de Transcrição E2F3/metabolismo , Feminino , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Neurônios/citologia , Neurônios/metabolismo , Proteína do Retinoblastoma/genética
16.
In Vitro Cell Dev Biol Anim ; 54(4): 304-310, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29508126

RESUMO

The development of skeletal muscle is a complex process involving the proliferation, differentiation, apoptosis, and changing of muscle fiber types in myoblasts. Many reports have described the involvement of microRNAs in the myogenesis of myoblasts. In this study, we found that the expression of miR-152 was gradually down-regulated during myoblast proliferation, but gradually up-regulated during the differentiation of myoblasts. Transfection with miR-152 mimics restrained cell proliferation and decreased the expression levels of cyclin E, CDK4, and cyclin D1, but promoted myotube formation and significantly increased the mRNA expression levels of MyHC, MyoD, MRF4, and MyoG in C2C12 myoblasts. However, treatment with miR-152 inhibitors promoted cell proliferation and restrained differentiation. Moreover, over-expression of miR-152 significantly decreased E2F3 production in C2C12 myoblasts. A luciferase assay confirmed that miR-152 could bind to the 3' UTR of E2F3. In conclusion, this study showed that miR-152 inhibited proliferation and promoted myoblast differentiation by targeting E2F3.


Assuntos
Fator de Transcrição E2F3/genética , MicroRNAs/fisiologia , Mioblastos/citologia , Animais , Diferenciação Celular/genética , Proliferação de Células/genética , Fator de Transcrição E2F3/metabolismo , Feminino , Regulação da Expressão Gênica , Células HeLa , Humanos , Camundongos Endogâmicos , MicroRNAs/antagonistas & inibidores , MicroRNAs/genética , Fibras Musculares Esqueléticas/metabolismo , Mioblastos/metabolismo
17.
Biol Psychiatry ; 84(3): 167-179, 2018 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-29397901

RESUMO

BACKGROUND: Lasting changes in gene expression in brain reward regions, including nucleus accumbens (NAc), contribute to persistent functional changes in the addicted brain. We and others have demonstrated that altered expression of several candidate transcription factors in NAc regulates drug responses. A recent large-scale genome-wide study from our group predicted transcription factor E2F3 (E2F3) as a prominent upstream regulator of cocaine-induced changes in gene expression and alternative splicing. METHODS: We studied expression of two E2F3 isoforms-E2F3a and E2F3b-in mouse NAc after repeated cocaine administration and assayed the effects of overexpression or depletion of E2f3 isoforms in NAc on cocaine behavioral responses. We then performed RNA sequencing to investigate the effect of E2f3a overexpression in this region on gene expression and alternative splicing and performed quantitative chromatin immunoprecipitation at downstream targets in NAc following E2f3a overexpression or repeated cocaine exposure. Sample sizes varied between experiments and are noted in the text. RESULTS: We showed that E2f3a, but not E2f3b, overexpression or knockdown in mouse NAc regulates cocaine-induced locomotor and place conditioning behavior. Furthermore, we demonstrated that E2f3a overexpression substantially recapitulates genome-wide transcriptional profiles and alternative splicing induced by cocaine. We further validated direct binding of E2F3a at key target genes following cocaine exposure. CONCLUSIONS: This study establishes E2F3a as a novel transcriptional regulator of cocaine action in NAc. The findings reveal a crucial role for E2F3a in the regulation of cocaine-elicited behavioral states. Moreover, the importance of this role is bolstered by the extensive recapitulation of cocaine's transcriptional effects in NAc by overexpression of E2f3a.


Assuntos
Processamento Alternativo , Cocaína/farmacologia , Fator de Transcrição E2F3/fisiologia , Núcleo Accumbens/fisiologia , Animais , Comportamento Animal , Imunoprecipitação da Cromatina , Fator de Transcrição E2F3/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Núcleo Accumbens/efeitos dos fármacos , Isoformas de Proteínas/genética
18.
J Cancer Res Clin Oncol ; 144(3): 531-542, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29350287

RESUMO

PURPOSE: lncRNA H19 has been considered as an oncogenic lncRNA in many human tumours. In the present study, we identify the role and molecular mechanism of lncRNA H19 in melanoma. METHOD: QRT-PCR was used to detect the expression of lncRNA H19 and E2F3 was detected in melanoma tissues. Cell counting kit-8 (CCK8), representative metabolites analysis was used to explore the biological function of lncRNA H19, miR-106a-5p and E2F3 in melanoma cells. Bioinformatics, luciferase reporter assays, MS2-RIP and RNA pull-down assay was used to demonstrate the molecular mechanism of lncRNA H19 in melanoma. We further test the function of lncRNA H19 in vivo though Xenograft tumour assay. RESULTS: We found that lncRNA H19 was increased in melanoma tissue, and lncRNA H19 was correlated with poor prognosis of melanoma patients. miR-106a-5p acts as a tumour suppressor in melanoma by targeting E2F3. E2F3 affects the melanoma cell glucose metabolism and growth. We also demonstrated that lncRNA H19 may function as the sponge of miR-106a-5p to up-regulate E2F3 expression, and consequently promote the glucose metabolism and growth of melanoma. CONCLUSIONS: This result elucidates a new mechanism for lncRNA H19 in melanoma development and provides a survival indicator and potential therapeutic target for melanoma patients.


Assuntos
Proliferação de Células/genética , Fator de Transcrição E2F3/fisiologia , Glucose/metabolismo , Melanoma , MicroRNAs/fisiologia , RNA Longo não Codificante/fisiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Metabolismo dos Carboidratos/genética , Linhagem Celular Tumoral , Fator de Transcrição E2F3/genética , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Masculino , Melanoma/genética , Melanoma/metabolismo , Melanoma/mortalidade , Melanoma/patologia , Camundongos , Camundongos Nus , MicroRNAs/genética , Pessoa de Meia-Idade , Transdução de Sinais/genética , Neoplasias Cutâneas/genética , Neoplasias Cutâneas/metabolismo , Neoplasias Cutâneas/patologia
19.
J Biol Chem ; 293(9): 3156-3167, 2018 03 02.
Artigo em Inglês | MEDLINE | ID: mdl-29330306

RESUMO

E2F transcription factors are regulated by binding to the retinoblastoma (Rb) tumor suppressor family of proteins. Previously, we reported an E2FLQ mutation that disrupts the binding with Rb proteins without affecting the transcriptional activity of E2F. We also showed that mouse embryonic fibroblasts with an E2F3LQ mutation exhibit increased E2F activity and more rapid cell proliferation. In this report, we analyzed E2F3LQ mice to further characterize the in vivo consequences of Rb family-independent E2F3 activity. We found that homozygous E2F3LQ mice were viable and had no obvious developmental defects or tumor growth. Our results also indicated that E2F3LQ cells largely retain normal control of cell proliferation in vivo However, female E2F3LQ mice had partial nursing defects. Examination of the E2F3LQ mammary glands revealed increased caveolin-1 (CAV1) expression, reduced prolactin receptor/Stat5 signaling, and impaired pregnancy-induced cell proliferation and differentiation. Of note, ChIP experiments disclosed that E2F3 binds the CAV1 promoter. Furthermore, E2F3 overexpression induced CAV1 expression, and CRISPR/CAS9-mediated E2F3 knockout reduced CAV1 levels and also increased prolactin receptor-induced Stat5 signaling in mammary epithelial cells. Our results suggest that the Rb family-independent E2F3 LQ variant inhibits pregnancy-induced mammary gland cell proliferation and differentiation by up-regulating CAV1 expression and inhibiting Stat5 signaling.


Assuntos
Fator de Transcrição E2F3/genética , Fator de Transcrição E2F3/metabolismo , Glândulas Mamárias Animais/citologia , Mutação , Fator de Transcrição STAT5/metabolismo , Transdução de Sinais , Animais , Sequência de Bases , Caveolina 1/genética , Diferenciação Celular/genética , Proliferação de Células/genética , Feminino , Regulação da Expressão Gênica/genética , Camundongos , Gravidez
20.
J Cell Biochem ; 119(4): 3429-3439, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29135049

RESUMO

E2F3 is a transcription factor that has been shown to be overexpressed in hepatocellular carcinoma (HCC). It is well-known that the E2F3 gene encodes two proteins E2F3a and E2F3b. Therefore, the functions of the two distinct isoforms need to be clarified separately. To characterize the function of E2F3b in HCC, the effects of ectopic expression of E2F3b on cell proliferation, cell cycle, apoptosis and gene expression were investigated. E2F3b promoted G1/S phase transition and markedly increased cell proliferation, but had minor effect on apoptosis. Microarray analyses identified 366 differentially expressed genes (171 upregulated and 195 downregulated) in E2F3b- overexpressing cells. Differential expression of 16 genes relevant to cell cycle and cell proliferation were further verified by real-time PCR. Six genes, including CDC2, CCNE1, ARF, MAP4K2, MUSK, and PAX2 were confirmed to be upregulated by more than twofold; one gene, CCNA2 was validated to be downregulated by more than twofold. We also confirmed that E2F3b increased the protein levels of both cyclin E and Arf but did not affect cyclin D1 protein. These results suggest that E2F3b functions as an important promoter for cell proliferation and plays important roles in transcriptional regulation in HepG2 liver cancer cells.


Assuntos
Fator de Transcrição E2F3/genética , Fator de Transcrição E2F3/metabolismo , Perfilação da Expressão Gênica/métodos , Neoplasias Hepáticas/genética , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Processamento Alternativo , Apoptose , Ciclo Celular , Proliferação de Células , Regulação Neoplásica da Expressão Gênica , Redes Reguladoras de Genes , Células Hep G2 , Humanos , Neoplasias Hepáticas/metabolismo
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