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1.
Aging (Albany NY) ; 13(3): 3866-3885, 2021 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-33461166

RESUMO

Long non-coding RNAs (lncRNA) play a vital role in colorectal cancer (CRC) progression. To investigate the role of long intergenic non-coding RNA LINC00485 in CRC, we performed in vitro functional experiments. LoVo tumor-bearing and liver metastasis mice were used as in vivo models. We found that LINC00485 expression was significantly lower in CRC tissues and cancer cells than in paired normal samples and human normal colonic epithelial cells. Lower expression of LINC00485 predicted poor prognosis in CRC patients. LINC00485 knockdown promoted the proliferation, migration, and invasion of FHC cells, while LINC00485 overexpression weakened these abilities of LoVo cells. MicroRNA miR-581 was the downstream target of LINC00485, which was downregulated in CRC samples and cancer cells compared to normal tissues and normal colonic epithelial cells. MiR-581 overexpression induced proliferation, migration, and invasion of FHC cells, while miR-581 antagomir treatment produced opposite results. MiR-581 directly targeted the 3'UTR of EDEM1 and inhibited its expression and induction of epithelial-mesenchymal transition of CRC. In mouse models, LINC00485 knockdown or down-regulation of miR-581 significantly repressed CRC cell growth and prevented CRC liver metastasis. Overall, LINC00485 suppressed CRC tumorigenesis and progression by targeting the miR-581/EDEM1 axis. LINC00485 may be a potential therapeutic target for CRC.

2.
Mol Ther Nucleic Acids ; 22: 750-765, 2020 Dec 04.
Artigo em Inglês | MEDLINE | ID: mdl-33230473

RESUMO

Hepatocellular carcinoma (HCC), one of the most aggressive malignancies, ranks as the fourth leading cause of cancer-related deaths worldwide. Emerging evidence indicates that RNA N6-methyladenosine (m6A) plays a critical role in tumor progression. However, the biological function of YTHDF1 in HCC remains unclear. Here, we found that YTHDF1 expression was strikingly elevated in HCC tissues and cell lines and significantly associated with prognosis of HCC patients. Moreover, YTHDF1 expression was transcriptionally regulated by USF1 and c-MYC in HCC. Functional studies showed that YTHDF1 can promote HCC cell proliferation and metastasis both in vitro and in vivo. Multi-omics analysis revealed that YTHDF1 can accelerate the translational output of FZD5 mRNA in an m6A-dependent manner and function as an oncogene through the WNT/ß-catenin pathway. Taken together, our study revealed an essential role of YTHDF1 in the progression of HCC cells, which indicated that targeting YTHDF1 may be a potential therapeutic strategy in HCC.

3.
Aging (Albany NY) ; 12(21): 21638-21659, 2020 11 06.
Artigo em Inglês | MEDLINE | ID: mdl-33159022

RESUMO

N6-methyladenosine (m6A) RNA methylation is the most prevalent modification of messenger RNAs (mRNAs) and catalyzed by a multicomponent methyltransferase complex (MTC), among which methyltransferase-like 3 (METTL3) and METTL14 are two core molecules. However, METTL3 and METTL14 play opposite regulatory roles in hepatocellular carcinoma (HCC). Based on The Cancer Genome Atlas (TCGA) database and Gene Expression Omnibus (GEO) database, we conducted a multi-omics analysis of METTL3 and METTL14 in HCC, including RNA-sequencing, m6ARIP-sequencing, and ribosome-sequencing profiles. We found that the expression and prognostic value of METTL3 and METTL14 are opposite in HCC. Besides, after METTL3 and METTL14 knockdown, most of the dysregulated mRNAs, signaling pathways and biological processes are distinct in HCC, which partly explains the contrary regulatory role of METTL3 and METTL14. Intriguingly, these mRNAs whose stability or translation efficiency are influenced by METTL3 or METTL14 in an m6A dependent manner, jointly regulate multiple signaling pathways and biological processes, which supports the cooperative role of METTL3 and METTL14 in catalyzing m6A modification. In conclusion, our study further clarified the contradictory role of METTL3 and METTL14 in HCC.

4.
Mol Cancer ; 19(1): 106, 2020 06 17.
Artigo em Inglês | MEDLINE | ID: mdl-32552762

RESUMO

BACKGROUND: Colorectal cancer (CRC) is one of the leading causes of tumor-related death worldwide, and its main cause of death is distant metastasis. Methyltransferase-like 14(METTL14), a major RNA N6-adenosine methyltransferase, is involved in tumor progression via regulating RNA function. The goal of the study is to uncover the biological function and molecular mechanism of METTL14 in CRC. METHODS: Quantitative real-time PCR (qRT-PCR), western blot and immunohistochemical (IHC) assays were employed to detect METTL14 and SOX4 in CRC cell lines and tissues. The biological functions of METTL14 were demonstrated using in vitro and in vivo experiments. Chromatin immunoprecipitation (ChIP), Transcrptomic RNA sequencing (RNA-Seq), m6A-RNA immunoprecipitation sequencing (MeRIP-Seq), RNA immunoprecipitation and luciferase reporter assays were used to explore the mechanism of METTL14 action. RESULTS: METTL14 expression was significantly downregulated in CRC and decreased METTL14 was associated with poor overall survival (OS). Both the univariate and multivariate Cox regression analysis indicated that METTL14 was an independent prognostic factor in CRC. Moreover, lysine-specific histone demethylase 5C(KDM5C)-mediated demethylation of histone H3 lysine 4 tri-methylation(H3K4me3) in the promoter of METTL14 inhibited METTL14 transcription. Functionally, we verified that METTL14 inhibited CRC cells migration, invasion and metastasis through in vitro and in vivo assays, respectively. Furthermore, we identified SRY-related high-mobility-group box 4(SOX4) as a target of METTL14-mediated m6A modification. Knockdown of METTL14 markedly abolished SOX4 mRNA m6A modification and elevated SOX4 mRNA expression. We also revealed that METTL14-mediated SOX4 mRNA degradation relied on the YTHDF2-dependent pathway. Lastly, we demonstrated that METTL14 might inhibit CRC malignant process partly through SOX4-mediated EMT process and PI3K/Akt signals. CONCLUSIONS: Decreased METTL14 facilitates tumor metastasis in CRC, suggesting that METTL14 might be a potential prognostic biomarker and effective therapeutic target for CRC.

5.
Ann Biomed Eng ; 2020 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-32504141

RESUMO

This paper reports a new type of augmented reality (AR) system that integrates a Microsoft HoloLens device with a three-dimensional (3D) point tracking module for medical training and telementored surgery. In this system, a stereo camera is used to track the 3D position of a scalpel and transfer its coordinates wirelessly to a HoloLens device. In the scenario of surgical training, a virtual surgical scene with pre-recorded surgical annotations is superimposed with the actual surgical scene so that the surgical trainee is able to operate following virtual instructions. In the scenario of telementored surgery, the virtual surgical scene is co-registered with the actual surgical scene so that the virtual scalpel remotely mentored by an experienced surgeon provides the AR guidance for the inexperienced on-site operator. The performance characteristics of the proposed AR telementoring system are verified by benchtop experiments. The clinical applicability of the proposed system in telementored skin grafting surgery and fasciotomy is validated in a New Zealand rabbit model. Our benchtop and in vivo experiments demonstrate the potential to improve surgical performance and reduce healthcare disparities in remote areas with limited resources.

6.
IUBMB Life ; 72(7): 1393-1403, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32107843

RESUMO

Metastasis is responsible for 90% of colorectal cancer (CRC)-related deaths. In the present study, we identified a novel key regulator of CRC metastasis, leucine-rich repeats and immunoglobulin-like domains protein 3 (LRIG3), which was significantly decreased in CRC tissues and cell lines. Downregulation of LRIG3 was attributed to copy number loss and promoter hypermethylation. Low LRIG3 expression was positively correlated with metastatic clinical features and shorter survival time. Functional experiments showed that knockout of LRIG3 markedly enhanced CRC cell migration and invasion ability, whereas reintroduction of LRIG3 exerted the opposite effects. Regarding the mechanism, LRIG3 could facilitate the binding of DUSP6 to ERK1/2, resulting in the dephosphorylation of ERK1/2 and subsequently downregulation of slug, an epithelial-to-mesenchymal transition trigger, thereby constraining CRC cell motility. Importantly, LRIG3 expression was strongly negatively correlated with slug or p-ERK1/2 expression in CRC tissues. Collectively, our data suggest that LRIG3 is a novel suppressor of CRC metastasis, reactivation of LRIG3 may be a promising therapeutic approach for metastatic CRC patients.

7.
J Biomed Opt ; 24(8): 1-4, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31436070

RESUMO

We propose a handheld projective imaging device for orthotopic projection of near-infrared fluorescence images onto target biological tissue at visible wavelengths without any additional visual aid. The device integrates a laser diode light source module, a camera module, a projector, an ultrasonic distance sensor, a Raspberry Pi single-board computer, and a battery module in a rugged handheld unit. It is calibrated at the detected working distance for seamless coregistration between fluorescence emission and projective imaging at the target tissue site. The proposed device is able to achieve a projection resolution higher than 314 µm and a planar projection bias less than 1 mm at a projection field of view of 58 × 108 mm2 and a working distance of 27 cm. Technical feasibility for projective imaging is verified in an ex vivo model of chicken breast tissue using indocyanine green as a fluorescence agent. Clinical utility for image-guided surgery is demonstrated in a clinical trial where sentinel lymph nodes in breast cancer patients are identified and resected under the guidance of projective imaging. Our ex vivo and in vivo experiments imply the clinical utility of deploying the proposed device for image-guided surgical interventions in resource-limited settings.


Assuntos
Neoplasias da Mama/diagnóstico por imagem , Neoplasias da Mama/cirurgia , Biópsia de Linfonodo Sentinela/métodos , Espectrometria de Fluorescência/instrumentação , Espectroscopia de Luz Próxima ao Infravermelho/instrumentação , Cirurgia Assistida por Computador , Desenho de Equipamento , Feminino , Fluorescência , Humanos , Verde de Indocianina , Lasers , Metástase Linfática , Imagem Óptica , Linfonodo Sentinela/patologia
8.
Neuropsychiatr Dis Treat ; 14: 3121-3132, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30532543

RESUMO

Background: Neuropsychiatric disorders are devastating illnesses worldwide; however, the potential involvement of viruses in the pathophysiological mechanisms of psychiatric diseases have not been clearly elucidated. Borna disease virus (BDV) is a neurotropic, noncytopathic RNA virus. Materials and methods: In this study, we infected neonatal rats intracranially with BDV Hu-H1 and Strain V within 24 hours of birth. Psychological phenotypes were assessed using sucrose preference test, open field test, elevated plus maze test, and forced swim test. The protein expression of ERK/CREB/BDNF pathway was assessed by Western blotting of in vitro and in vivo samples. Results: Hu-H1-infected rats showed anxiety-like behavior 8 weeks postinfection while Strain V-infected rats demonstrated a certain abnormal behavior. Phosphorylated ERK1/2 was significantly upregulated in the hippocampi of Strain V- and Hu-H1-infected rats compared with control rats, indicating that Raf/MEK/ERK signaling was activated. Conclusion: The data suggested that infection of neonatal rats with BDV Hu-H1 and Strain V caused behavioral abnormalities that shared common molecular pathways, providing preliminary evidences to investigate the underlying mechanisms of psychiatric disorders caused by BDV.

9.
Biotechnol Adv ; 36(8): 2129-2137, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30290194

RESUMO

Since its discovery in the 1970s, the T7 RNA polymerase (T7 RNAP) transcription system has been applied extensively as an effective tool in molecular biology because of its robust function in various hosts, including prokaryotic, eukaryotic and cell free systems. Recently, the T7 RNAP transcription system has emerged as a critical component for synthetic biology. The present paper summarizes the advances of the T7 RNAP transcription system in synthetic biology, including the recent progress of T7 RNAP structure and its cognate promoter and terminator and its application in cell free systems, logic gates and orthogonal genetic circuits.


Assuntos
Bacteriófago T7/genética , Regiões Promotoras Genéticas/genética , Biologia Sintética/métodos , DNA Viral/genética , Engenharia Genética , Transcrição Genética/genética
10.
Life Sci ; 211: 102-117, 2018 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-30201296

RESUMO

AIMS: Although decades of research have revealed numerous molecular abnormalities in the hippocampus associated with depression, the different mechanisms involved in the susceptibility and resilience of mice to chronic social defeat stress (CSDS)-induced depression remain poorly understand. Through the social defeat model, we can study the differences in molecular changes between the susceptible and resilient mice. MAIN METHODS: We used a proteomic-based platform to compare hippocampal proteins in CSDS mice with those in control mice. Differentially expressed proteins were identified through isobaric tags for relative and absolute quantitation (iTRAQ) combined with LC-MS/MS. We then analyzed the results by ingenuity pathway analysis (IPA) and verified five proteins by western blotting. KEY FINDINGS: Mice were exposed to 10 days of CSDS, which successfully induced stress-susceptible and -resilient phenotypes. 161 and 134 proteins were significantly differentially expressed in the susceptible and resilient groups, respectively, compared with the levels in the control group. The Rac signaling and the GABA receptors signaling pathways were the common top-ranking pathways. We found that low-density lipoprotein receptor-related protein 6 (LRP6) was upregulated in resilient mice and neuropeptide Y (NPY) was downregulated in susceptible mice compared with the levels in control mice. Moreover, neuropeptide Y receptor type 2 (NPY2R) protein expression in susceptible mice was downregulated compared with that in the resilient group. SIGNIFICANCE: Our findings in the three groups potentially reveal the differences in molecular mechanisms underlying depression between susceptible and resilient mice. The results provide insight into molecular abnormalities of the hippocampus in CSDS mice and some potential drug targets for treating depression.


Assuntos
Transtorno Depressivo/patologia , Suscetibilidade a Doenças , Hipocampo/patologia , Proteína-6 Relacionada a Receptor de Lipoproteína de Baixa Densidade/metabolismo , Neuropeptídeo Y/metabolismo , Proteômica/métodos , Receptores de Neuropeptídeo Y/metabolismo , Estresse Psicológico , Animais , Transtorno Depressivo/metabolismo , Modelos Animais de Doenças , Hipocampo/metabolismo , Marcação por Isótopo/métodos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Comportamento Social
11.
Cell Physiol Biochem ; 49(1): 381-394, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30138929

RESUMO

BACKGROUND/AIMS: Borna disease virus 1 (BoDV-1) infection induces cognitive impairment in rodents. Emerging evidence has demonstrated that Chromatin remodeling through histone acetylation can regulate cognitive function. In the present study, we investigated the epigenetic regulation of chromatin that underlies BoDV-1-induced cognitive changes in the hippocampus. METHODS: Immunofluorescence assay was applied to detect BoDV-1 infection in hippocampal neurons and Sprague-Dawley rats models. The histone acetylation levels both in vivo and vitro were assessed by western blots. The acetylation-regulated genes were identified by ChIP-seq and verified by RT-qPCR. Cognitive functions were evaluated with Morris Water Maze test. In addition, Golgi staining, and electrophysiology were used to study changes in synaptic structure and function. RESULTS: BoDV-1 infection of hippocampal neurons significantly decreased H3K9 histone acetylation level and inhibited transcription of several synaptic genes, including postsynaptic density 95 (PSD95) and brain-derived neurotrophic factor (BDNF). Furthermore, BoDV-1 infection of Sprague Dawley rats disrupted synaptic plasticity and caused spatial memory impairment. These rats also exhibited dysregulated hippocampal H3K9 acetylation and decreased PSD95 and BDNF protein expression. Treatment with the HDAC inhibitor, suberanilohydroxamic acid (SAHA), attenuated the negative effects of BoDV-1. CONCLUSION: Our results demonstrate that regulation of H3K9 histone acetylation may play an important role in BoDV-1-induced memory impairment, whereas SAHA may confer protection against BoDV-1-induced cognitive impairments. This study finds important mechanism of BoDV-1 infection disturbing neuronal synaptic plasticity and inducing cognitive dysfunction from the perspective of histone modification.


Assuntos
Doença de Borna/patologia , Vírus da Doença de Borna/fisiologia , Histonas/metabolismo , Memória/fisiologia , Acetilação/efeitos dos fármacos , Animais , Doença de Borna/virologia , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Células Cultivadas , Imunoprecipitação da Cromatina , Modelos Animais de Doenças , Proteína 4 Homóloga a Disks-Large/metabolismo , Ácidos Hidroxâmicos/farmacologia , Aprendizagem em Labirinto , Memória/efeitos dos fármacos , Plasticidade Neuronal/genética , Neurônios/citologia , Neurônios/metabolismo , Ratos , Ratos Sprague-Dawley , Vorinostat
12.
ACS Synth Biol ; 7(5): 1424-1435, 2018 05 18.
Artigo em Inglês | MEDLINE | ID: mdl-29609457

RESUMO

Metabolic engineering and synthetic biology usually require universal expression systems for stable and efficient gene expression in various organisms. In this study, a host-independent and stable T7 expression system had been developed by integrating T7 RNA polymerase and its cognate transcriptional units in single plasmid. The expression of T7 RNA polymerase was restricted below its lethal threshold using a T7 RNA polymerase antisense gene cassette, which allowed long periods of cultivation and protein production. In addition, by designing ribosome binding sites, we further tuned the expression capacity of this novel T7 system within a wide range. This host-independent expression system efficiently expressed genes in five different Gram-negative strains and one Gram-positive strain and was also shown to be applicable in a real industrial d- p-hydroxyphenylglycine production system.


Assuntos
RNA Polimerases Dirigidas por DNA/genética , Glicina/análogos & derivados , Engenharia Metabólica/métodos , Plasmídeos/genética , Proteínas Virais/genética , Elementos Antissenso (Genética) , Sítios de Ligação , Corynebacterium glutamicum/genética , RNA Polimerases Dirigidas por DNA/metabolismo , Regulação Bacteriana da Expressão Gênica , Glicina/biossíntese , Bactérias Gram-Negativas/genética , Microrganismos Geneticamente Modificados , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Ribossomos/metabolismo , Sinorhizobium/genética , Transcrição Genética , Proteínas Virais/metabolismo
13.
Neuropharmacology ; 128: 119-131, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-28986280

RESUMO

OBJECTIVES: Recombinant tissue plasminogen activator (rtPA) is widely used for patients with thromboembolic disease, and increasing evidence indicates that it can directly induce neurotoxicity independent of its thrombolysis property. Here, we aimed to confirm the long-term effect of rtPA on animal's behavior, and investigate the underlying pathogenesis. METHODS AND RESULTS: Male Sprague-Dawley rats randomly received a dose of rtPA (10 mg/kg) or sterile saline. Three months later, the animals receiving rtPA displayed anxiety-like behaviors in the open field and novelty-suppressed feeding tests. To investigate the possible pathogenesis, gas chromatography-mass spectrometry-based metabolomics analysis was performed, with 18 differential metabolites identified in the hippocampus 24 h after the treatments. Based upon these differential metabolites, a metabolite-protein integrated network was generated, which indicated that ERK1/2-glutamic acid decarboxylase (GAD) 1-γ aminobutyric acid (GABA) cascade may be related to long-term anxiety-like behaviors. The GABA levels in hippocampus were decreased 24 h post-treatment and three months later, confirmed by a high performance liquid chromatography method. We also examined the expression of GAD1 and GAD2 using western blotting or immunohistochemical staining. Levels of GAD1 were persistently decreased after treatment, while GAD2 levels, GAD1-immunoreactive, and GAD2-immunoreactive neurons showed no significant differences. The underlying pathogenesis also involved activation of ERK1/2, confirmed by increased phospho-ERK1/2 24 h post-treatment. CONCLUSIONS: RtPA can induce long-term anxiety-like behaviors after a clinical injected dose. The underlying pathogenesis involves the ERK1/2-GAD1-GABA cascade in the hippocampus. This pharmacological side effect of rtPA may further exacerbate post-stroke anxiety disorder for stroke patients.


Assuntos
Ansiedade/induzido quimicamente , Ansiedade/patologia , Hipocampo/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Ativador de Plasminogênio Tecidual/toxicidade , Animais , Peso Corporal/efeitos dos fármacos , Modelos Animais de Doenças , Comportamento Exploratório/efeitos dos fármacos , Comportamento Alimentar/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Glutamato Descarboxilase/metabolismo , Elevação dos Membros Posteriores , Hipocampo/metabolismo , Sistema de Sinalização das MAP Quinases/fisiologia , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Redes e Vias Metabólicas/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Ratos , Ratos Sprague-Dawley , Proteína X Associada a bcl-2/metabolismo , Ácido gama-Aminobutírico/metabolismo
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