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1.
Gene ; 725: 144143, 2020 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-31629816

RESUMO

Atherosclerosis is a common cardiovascular disorder and is characterized by damage of endothelial cells, cell inflammation, hyper-proliferation of vascular smooth muscle cells and the accumulation of extracellular lipids and fibrous tissues. In this study, we firstly examined the expression level of long intergenic non-protein coding RNA, regulator of reprogramming (linc-ROR) in homocysteine (Hcy)-stimulated human aortic smooth muscle cells (HASMCs), and then looked into the potential molecular signaling axis of linc-ROR in regulating the proliferation and migration of HASMCs. Hcy promoted HASMC proliferation and up-regulated linc-ROR expression. Functional studies showed that linc-ROR exerted enhanced actions on the proliferation and migration of HASMCs. In addition, linc-ROR acted as a competing endogenous RNA for miR-195-5p and repressed the miR-195-5p expression in HASMCs. Linc-ROR was up-regulated the miR-195-3p was down-regulated in the plasma from CAD patients when compared to normal controls. Furthermore, fibroblast growth factor 2 (FGF2) was identified as a target of miR-195-5p and was negatively regulated by miR-195-5p in HASMCs. The rescue experiments revealed that linc-ROR-mediated HASMC proliferation and migration may be via regulating miR-195-5p/FGF2 axis. Linc-ROR inhibition blocked the miR-195-5p/FGF2 signaling in Hcy-treated HASMCs, and this effect may also involve in the miR-195-5p/FGF2 axis. To summarize, the data of the present study identified the up-regulation of linc-ROR in Hcy-stimulated HASMCs, and further mechanistic functional studies revealed that linc-ROR promoted HASMC proliferation and migration via regulating miR-195-5p/FGF2 axis. The present study provided the novel actions of linc-ROR in regulating HASMC proliferation and migration, which may be related to the pathophysiology of atherosclerosis.


Assuntos
Fator 2 de Crescimento de Fibroblastos/metabolismo , MicroRNAs/metabolismo , RNA Longo não Codificante/metabolismo , Aorta/citologia , Aorta/efeitos dos fármacos , Aorta/metabolismo , Aterosclerose/genética , Aterosclerose/metabolismo , Aterosclerose/patologia , Autofagia/fisiologia , Linhagem Celular Tumoral , Movimento Celular/fisiologia , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/fisiologia , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/metabolismo , Fator 2 de Crescimento de Fibroblastos/genética , Fator 2 de Crescimento de Fibroblastos/fisiologia , Homocisteína/farmacologia , Humanos , MicroRNAs/genética , Miócitos de Músculo Liso/citologia , Miócitos de Músculo Liso/efeitos dos fármacos , Miócitos de Músculo Liso/metabolismo , RNA Longo não Codificante/biossíntese , RNA Longo não Codificante/genética , Transdução de Sinais
2.
Medicine (Baltimore) ; 98(32): e16470, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31393351

RESUMO

We aimed to investigate the correlation of long noncoding RNA nuclear enriched abundant transcript 1 (lnc-NEAT1), microRNA-124 (miR-124) and lnc-NEAT1/miR-124 axis with disease risk, severity, inflammatory cytokines, and survival of sepsis.Eighty-two patients with sepsis and 82 healthy controls (HCs) were consecutively enrolled. Blood samples were collected for detection of lnc-NEAT1 and miR-124 expressions (using RT-qPCR) and measurement of inflammatory cytokines expressions (by ELISA). Severity and organ failure were assessed by acute physiology and chronic health evaluation II (APACHE II) score and sequential organ failure assessment (SOFA) score, and survival was assessed.Lnc-NEAT1 expression was increased while miR-124 expression was decreased in patients with sepsis compared to HCs, and both of them were able to distinguish patients with sepsis from HCs. For disease condition, lnc-NEAT1 positively associated with APACHE II score, SOFA score, and expressions of C-reactive protein (CRP), procalcitonin, tumor necrosis factor α (TNF-α), and interleukin-1ß (IL-1ß), whereas miR-124 negatively correlated with APACHE II score, SOFA score and levels of serum creatinine (Scr), CRP, TNF-α, IL-1ß, interleukin-6 (IL-6) and interleukin-17 (IL-17). Regarding prognosis, lnc-NEAT1 was upregulated but miR-124 was downregulated in nonsurvivors compared to survivors. Additionally, lnc-NEAT1 negatively correlated with miR-124. Besides, lnc-NEAT1/miR-124 axis was increased in patients with sepsis compared to HCs, and positively associated with APACHE II score, SOFA score, and levels of Scr, CRP, TNF-α, IL-1ß, IL-6, and IL-17, while negatively correlated with survival. Most importantly, lnc-NEAT1/miR-124 axis presented numerically increased predictive value for sepsis risk and survival compared to each index alone.Lnc-NEAT1/miR-124 axis correlates with increased sepsis risk, and associates with higher inflammation, deteriorative disease condition, and decreased survival in patients with sepsis.


Assuntos
Mediadores da Inflamação/metabolismo , RNA Longo não Codificante/biossíntese , Sepse/mortalidade , Sepse/fisiopatologia , APACHE , Adulto , Idoso , Deterioração Clínica , Citocinas/metabolismo , Feminino , Humanos , Masculino , MicroRNAs/biossíntese , Pessoa de Meia-Idade , Escores de Disfunção Orgânica , Índice de Gravidade de Doença
3.
Gene ; 715: 144029, 2019 Oct 05.
Artigo em Inglês | MEDLINE | ID: mdl-31376409

RESUMO

Intervertebral disc degeneration (IDD) is a major cause of lower back pain, but the specific molecular mechanisms governing its development are poorly characterized. This study sought to assess to what extent HOTAIR, a long non-coding (Lnc) RNA is expressed in IDD and regulates the apoptotic death of nucleus pulposus (NP) cells. We therefore used real-time qPCR to measure HOTAIR and microRNA(miR)-34a-5p in degenerative NP cells, and then validated their functional relevance via overexpressing them in these NP cells. We further verified the targets of these RNA constructs in 293 T cells through the use of a dual luciferase reporter assay. We further measured NP cell apoptosis via flow cytometry and Notch1 expression via western blotting. Our results indicated that IDD was linked with decreased HOTAIR expression relative to regular NP cells, and overexpressing this lncRNA was linked to reduced apoptotic NP cell death, whereas overexpressing miR-34a-5p had the opposite effect. We found that HOTAIR served as a miR-34a-5p sponge, sequestering this miRNA and thereby down regulating genes linked to apoptosis through the Notch signaling pathway. Even in naturally degenerated NP cells, HOTAIR delayed the onset of apoptosis. Together these results reveal that a HOTAIR/miR-34a-5p/Notch1 signaling pathway may regulate the development of IDD, potentially making HOTAIR a viable target for treatment of this disease.


Assuntos
Apoptose , Regulação para Baixo , Degeneração do Disco Intervertebral/metabolismo , MicroRNAs/biossíntese , RNA Longo não Codificante/biossíntese , Receptor Notch1/biossíntese , Transdução de Sinais , Adulto , Idoso , Feminino , Humanos , Degeneração do Disco Intervertebral/genética , Degeneração do Disco Intervertebral/patologia , Masculino , MicroRNAs/genética , Pessoa de Meia-Idade , Núcleo Pulposo/patologia , RNA Longo não Codificante/genética , Receptor Notch1/genética
4.
Gene ; 715: 144012, 2019 Oct 05.
Artigo em Inglês | MEDLINE | ID: mdl-31357021

RESUMO

Long noncoding RNAs (lncRNAs) have been shown to play an important role in tumor biogenesis and prognosis. The glioma is a grade classified cancer, however, we still lack the knowledge on their function during glioma progression. While previous studies have shown how lncRNAs regulate protein-coding gene epigenetically, it is still unclear how lncRNAs are regulated epigenetically. In this study, we firstly analyzed the RNA-seq data systematically across grades II, IV, and IV of glioma samples. We identified 60 lncRNAs that are significantly differentially expressed over disease progression (DElncRNA), including well-known PVT1, HOTAIR, H19 and rarely studied CARD8-AS, MIR4435-2HG. Secondly, by integrating HM450K methylation microarray data, we demonstrated that some of the lncRNAs are epigenetically regulated by methylation. Thirdly, we developed a DESeq2-GSEA-ceRNA-survival analysis strategy to investigate their functions. Particularly, MIR4435-2HG is highly expressed in high-grade glioma and may have an impact on EMT and TNFα signaling pathway by functioning as a miRNA sponge of miR-125a-5p and miR-125b-5p to increase the expression of CD44. Our results revealed the dynamic expression of lncRNAs in glioma progression and their epigenetic regulation mechanism.


Assuntos
Metilação de DNA , DNA de Neoplasias , Epigênese Genética , Regulação Neoplásica da Expressão Gênica , Glioma , MicroRNAs , RNA Longo não Codificante , RNA Neoplásico , DNA de Neoplasias/genética , DNA de Neoplasias/metabolismo , Perfilação da Expressão Gênica , Glioma/genética , Glioma/metabolismo , Glioma/patologia , Humanos , MicroRNAs/biossíntese , MicroRNAs/genética , Proteínas de Neoplasias/biossíntese , Proteínas de Neoplasias/genética , RNA Longo não Codificante/biossíntese , RNA Longo não Codificante/genética , RNA Neoplásico/biossíntese , RNA Neoplásico/genética
5.
Hematol Oncol ; 37(4): 474-482, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31325181

RESUMO

LncRNAs play critical roles in various pathophysiological and biological processes, such as protein translation, RNA splicing, and epigenetic modification. Indeed, abundant evidences demonstrated that lncRNA act as competing endogenous RNAs (ceRNAs) to participate in tumorigenesis. However, little is known about the underlying function of lncRNA in nonhomologous end joining (NHEJ) pathway 1 (LINP1) in pediatric and adolescent acute myeloid leukemia (AML). The expression of LINP1 was examined in AML patient samples by qRT-PCR. Cell proliferation was examined by CCK-8 and Edu assays. ß-Galactosidase senescence assay, mGlucose uptake assay, lactate production assay, and Gene Ontology (GO) analysis were performed for functional analysis. We found that LINP1 was significantly overexpressed in AML patients at diagnosis, whereas downregulated after complete remission (CR). Furthermore, knockdown of LINP1 expression remarkably suppressed glucose uptake and AML cell maintenance. Mechanistically, LINP1 was found to inhibit the glucose metabolism by suppressing the expression of HNF4a. Both LINP1 and HNF4a knockdown reduced the expression levels of AMPK phosphorylation and WNT5A, indicating for the first time that LINP1 strengthened the HNF4a-AMPK/WNT5A signaling pathway involved in cell glucose metabolism modulation and AML cell survival. Taken together, our results indicated that LINP1 promotes the malignant phenotype of AML cells and stimulates glucose metabolism, which can be regarded as a potential prognostic marker and therapeutic target for AML.


Assuntos
Adenilato Quinase/fisiologia , Fator 4 Nuclear de Hepatócito/fisiologia , Leucemia Mieloide Aguda/genética , RNA Longo não Codificante/fisiologia , RNA Neoplásico/fisiologia , Transdução de Sinais/fisiologia , Proteína Wnt-5a/fisiologia , Adolescente , Animais , Medula Óssea/patologia , Divisão Celular , Criança , Regulação Leucêmica da Expressão Gênica , Técnicas de Silenciamento de Genes , Ontologia Genética , Glucose/metabolismo , Fator 4 Nuclear de Hepatócito/biossíntese , Fator 4 Nuclear de Hepatócito/genética , Humanos , Leucemia Mieloide Aguda/metabolismo , Leucemia Mieloide Aguda/patologia , Masculino , Camundongos , Camundongos Endogâmicos NOD , Camundongos SCID , Transplante de Neoplasias , Púrpura Trombocitopênica Idiopática/metabolismo , Interferência de RNA , RNA Longo não Codificante/biossíntese , RNA Longo não Codificante/genética , RNA Neoplásico/biossíntese , RNA Neoplásico/genética , RNA Interferente Pequeno/genética , Distribuição Aleatória , Indução de Remissão , Transdução de Sinais/genética , Células THP-1
6.
Oncol Rep ; 42(4): 1459-1466, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31322270

RESUMO

The expression of CDR1­AS, a representative circular RNA, is closely linked with poor prognosis in gastrointestinal cancers, such as colon, liver, and pancreatic cancers. Although it is well known that CDR1­AS antagonizes microRNA­7 function through its sequence similarities in the brain, its biological function and link with the malignant potential of cancer cells remain unclear, partly due to the difficulties of ectopic expression of circular RNAs. In the present study, SW620, a colon cancer cell line that stably expresses CDR1­AS RNA circularized, was established using the laccase 2 gene cassette, and its biological function associated with malignant behavior was determined. In contrast to previous studies, cell growth or invasion ability was not altered by CDR1­AS expression. However, the expression levels of CMTM4 and CMTM6, which were recently recognized as critical regulators of PD­L1 protein expression at the cell surface, were significantly increased. Accordingly, the cell surface PD­L1 protein levels were increased in CDR1­AS­expressing cells. Notably, the effects were not canceled out by overexpressing microRNA­7, indicating that the increase in cell surface PD­L1 in CDR1­AS­expressing cells was not dependent on microRNA­7 function. These results indicated that expression of this circular RNA in cancer cells may lead to poor prognosis by increasing cell surface PD­L1 levels through microRNA­7­independent mechanisms.


Assuntos
Antígeno B7-H1/biossíntese , Neoplasias Colorretais/metabolismo , RNA Longo não Codificante/biossíntese , Animais , Antígeno B7-H1/genética , Células CACO-2 , Processos de Crescimento Celular/fisiologia , Linhagem Celular Tumoral , Neoplasias Colorretais/genética , Neoplasias Colorretais/patologia , Células HEK293 , Humanos , Imuno-Histoquímica , Proteínas com Domínio MARVEL/biossíntese , Proteínas com Domínio MARVEL/genética , Masculino , Proteínas de Membrana/biossíntese , Proteínas de Membrana/genética , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , MicroRNAs/antagonistas & inibidores , MicroRNAs/genética , MicroRNAs/metabolismo , Invasividade Neoplásica , Prognóstico , RNA Longo não Codificante/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
7.
Life Sci ; 231: 116549, 2019 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-31200002

RESUMO

BACKGROUND: Long non-coding RNA (lncRNA) is emerging as an important regulator in various physiological and pathological processes. Recently, it was found that lncRNA long non-coding RNA tumor suppressor candidate 7 (TUSC7) could play tumor suppressive roles in several cancers. However, the function and underlying regulatory mechanism of lncRNA TUSC7 in endometrial carcinoma (EC) remains largely unclear. METHODS: The expression levels of TUSC7 and microRNAs-616 (miR-616) were analyzed by real-time PCR and in situ hybridization. Cell cycle and cell metastasis associated protein expressions were determined by western blotting. Cell proliferation, cycle and metastasis were determined by CCK-8 cell viability, colony formation, flow cytometer, wound scratch and transwell assays respectively in vitro. RNA pull-down, luciferase and western blotting assays were used to examine the target relationship between TUSC7 and miR-616 or that between miR-616 and suppressors of cytokine signaling 4 (5) (SOCS4 (SOCS5)). The functional effects of TUSC7 through sponging miR-616 were further examined using a xenograft tumor mouse model in vivo. RESULTS: TUSC7 was downexpressed in EC tissues and cell lines, and TUSC7 upregulation could remarkably inhibit cell proliferation, cycle progression and metastasis in EC cells. Mechanistic investigations demonstrated that TUSC7 can interact with miR-616 and decrease its expression, thereby upregulating the expression of miR-616's targets SOCS4 (SOCS5). Additionally, in vivo experiments using a xenograft tumor mouse model revealed that TUSC7 can serve as a tumor suppressor through sponging miR-616, and upregulating SOCS4 (SOCS5) in EC. CONCLUSIONS: In this study, a newly identified regulatory mechanism of lncRNA TUSC7/miR-616/ SOCS4 (SOCS5) axis was systematically studied, which may hold promise as a promising target for EC treatment.


Assuntos
Neoplasias do Endométrio/genética , Neoplasias do Endométrio/metabolismo , MicroRNAs/genética , RNA Longo não Codificante/metabolismo , Proteínas Supressoras da Sinalização de Citocina/metabolismo , Animais , Apoptose , Técnicas de Cultura de Células , Ciclo Celular , Linhagem Celular Tumoral , Movimento Celular/fisiologia , Proliferação de Células/fisiologia , Neoplasias do Endométrio/patologia , Feminino , Xenoenxertos , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , MicroRNAs/metabolismo , Pessoa de Meia-Idade , Invasividade Neoplásica , RNA Longo não Codificante/biossíntese , RNA Longo não Codificante/genética , Proteínas Supressoras da Sinalização de Citocina/genética
8.
BMC Genomics ; 20(1): 515, 2019 Jun 21.
Artigo em Inglês | MEDLINE | ID: mdl-31226932

RESUMO

BACKGROUND: Long non-coding RNAs (lncRNAs) perform crucial roles in biological process involving complex mechanisms. However, information regarding their abundance, characteristics and potential functions linked to fish skin color is limited. Herein, Illumina sequencing and bioinformatics were conducted on black, white, and red skin of Koi carp (Cyprinus carpio L.). RESULTS: A total of 590,415,050 clean reads, 446,614 putative transcripts, 4252 known and 72,907 novel lncRNAs were simultaneously obtained, including 92 significant differentially expressed lncRNAs and 722 mRNAs. Ccr_lnc5622441 and Ccr_lnc765201 were up-regulated in black and red skin, Ccr_lnc14074601 and Ccr_lnc2382951 were up-regulated in white skin, and premelanosome protein a (Pmela), Pmelb and tyrosinase (Tyr) were up-regulated in black skin. The expression patterns of 18 randomly selected differentially expressed genes were validated using the quantitative real-time PCR method. Moreover, 70 lncRNAs acting on 107 target mRNAs in cis and 79 lncRNAs acting on 41,625 target mRNAs in trans were investigated. The resulting co-expression networks revealed that a single lncRNA can connect with numerous mRNAs, and vice versa. To further reveal their potential functions, Gene Ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were analyzed, and membrane, pigment cell development, cAMP signaling, melanogenesis and tyrosine metabolism appear to affect skin pigmentation. Additionally, three lncRNAs (Ccr_lnc142711, Ccr_lnc17214525 and Ccr_lnc14830101) and three mRNAs (Asip, Mitf and Tyr) involved in the melanogenesis pathway were investigated in terms of potential functions in embryogenesis and different tissues. CONCLUSIONS: The findings broaden our understanding of lncRNAs and skin color genetics, and provide new insight into the mechanisms underlying lncRNA-mediated pigmentation and differentiation in Koi carp.


Assuntos
Carpas/genética , RNA Longo não Codificante/biossíntese , RNA Mensageiro/biossíntese , Pigmentação da Pele/genética , Animais , Carpas/anatomia & histologia , Regulação da Expressão Gênica , Sequenciamento de Nucleotídeos em Larga Escala/veterinária , Reação em Cadeia da Polimerase em Tempo Real
9.
J Clin Pathol ; 72(8): 513-519, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31154423

RESUMO

The importance of circular RNAs (circRNAs) in pathological processes like cancer is evident. Among the circRNAs, recent studies have brought circPVT1 under focus as the most potent oncogenic non-coding RNA. Recent studies on various aspects of circPVT1, including its biogenesis, molecular alteration and its probable role in oncogenesis, have been conducted for research and clinical interest. In this review, a first attempt has been made to summarise the available data on circPVT1 from PubMed and other relevant databases with special emphasis on its role in development, progression and prognosis of various malignant conditions. CircPVT1 is derived from the same genetic locus encoding for long non-coding RNA lncPVT1; however, existing literature suggested circPVT1 and lncPVT1 are transcripted independently by different promoters. The interaction between circRNA and microRNA has been highlighted in majority of the few malignancies in which circPVT1 was studied. Besides its importance in diagnostic and prognostic procedures, circPVT1 seemed to have huge therapeutic potential as evident from differential drug response of cancer cell line as well as primary tumors depending on expression level of the candidate. circPVT1 in cancer therapeutics might be promising as a biomarker to make the existing treatment protocol more effective and also as potential target for designing novel therapeutic intervention.


Assuntos
Biomarcadores Tumorais/genética , Neoplasias/genética , RNA/genética , Animais , Biomarcadores Tumorais/biossíntese , Transformação Celular Neoplásica/genética , Transformação Celular Neoplásica/metabolismo , Transformação Celular Neoplásica/patologia , Regulação Neoplásica da Expressão Gênica , Humanos , Técnicas de Diagnóstico Molecular , Neoplasias/metabolismo , Neoplasias/patologia , Neoplasias/terapia , Valor Preditivo dos Testes , Prognóstico , RNA/biossíntese , RNA Longo não Codificante/biossíntese , RNA Longo não Codificante/metabolismo
10.
Int J Oncol ; 55(1): 93-102, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31180520

RESUMO

Neuroblastoma (NB) is one of the most common extracranial solid tumors in children, which has complex molecular mechanisms. Increasing evidence has suggested that long noncoding RNAs (lncRNAs) account for NB pathogenesis. However, the function of small nucleolar RNA host gene 16 (SNHG16) in NB is currently unclear. In the present study, publically available data and clinical specimens were employed to verify the expression of SNHG16 in NB. Colony formation, real­time cell proliferation and migration assays were performed to demonstrate the status of cellular proliferation and migration. Flow cytometry was used to examine cell cycle progression in SH­SY5Y cells, and acridine orange/ethidium bromide staining and caspase­3/7 activity measurements were applied to study cell apoptosis. To explore the underlying mechanism of SNHG16 function, an online database was used to identify potential RNA­binding proteins that bind SNHG16. The expression of SNHG16 was revealed to be in line with the clinical staging of NB, and high SNHG16 expression was positively associated with poor clinical outcome. Furthermore, SNHG16 silencing inhibited cell proliferation, repressed migration, and induced cell cycle arrest at the G0/G1 phase in SH­SY5Y cells. Additionally, apoptosis was undetectable in SH­SY5Y cells following SNHG16 silencing. Bioinformatics analysis revealed that SNHG16 regulated cell proliferation in NB through transcriptional and translational pathways. These results suggested that SNHG16 may serve important roles in the development and progression of NB, and could represent a potential target for NB therapy.


Assuntos
Neuroblastoma/genética , RNA Longo não Codificante/genética , Apoptose/genética , Pontos de Checagem do Ciclo Celular/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Criança , Pré-Escolar , Inativação Gênica , Humanos , Lactente , Neuroblastoma/metabolismo , Neuroblastoma/patologia , Oncogenes , RNA Longo não Codificante/biossíntese , RNA Interferente Pequeno/administração & dosagem , RNA Interferente Pequeno/genética , Transfecção
11.
Appl Microbiol Biotechnol ; 103(15): 6107-6117, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31187211

RESUMO

Noncoding RNAs (ncRNAs), including microRNAs (miRNAs), small interfering RNAs (siRNAs), and long noncoding RNAs (lncRNAs), regulate target gene expression and can be used as tools for understanding biological processes and identifying new therapeutic targets. Currently, ncRNA molecules for research and therapeutic use are limited to ncRNA mimics made by chemical synthesis. We have recently established a high-yield and cost-effective method of producing bioengineered or biologic ncRNA agents (BERAs) through bacterial fermentation, which is based on a stable tRNA/pre-miR-34a carrier (~ 180 nt) that accommodates target small RNAs. Nevertheless, it remains a challenge to heterogeneously express longer ncRNAs (e.g., > 260 nt), and it is unknown if single BERA may carry multiple small RNAs. To address this issue, we hypothesized that an additional human pre-miR-34a could be attached to the tRNA/pre-miR-34a scaffold to offer a new tRNA/pre-miR-34a/pre-miR-34a carrier (~ 296 nt) for the accommodation of multiple small RNAs. We thus designed ten different combinatorial BERAs (CO-BERAs) that include different combinations of miRNAs, siRNAs, and antagomirs. Our data showed that all target CO-BERAs were successfully expressed in Escherichia coli at high levels, greater than 40% in total bacterial RNAs. Furthermore, recombinant CO-BERAs were purified to a high degree of homogeneity by fast protein liquid chromatography methods. In addition, CO-BERAs exhibited strong anti-proliferative activities against a variety of human non-small cell lung cancer cell lines. These results support the production of long ncRNA molecules carrying different warhead small RNAs for multi-targeting which may open avenues for developing new biologic RNAs as experimental, diagnostic, and therapeutic tools.


Assuntos
Antagomirs/biossíntese , Antagomirs/genética , Bioengenharia/métodos , RNA Longo não Codificante/biossíntese , RNA Longo não Codificante/genética , RNA Interferente Pequeno/biossíntese , RNA Interferente Pequeno/genética , Cromatografia Líquida , Escherichia coli/genética , Escherichia coli/metabolismo , Expressão Gênica , Humanos , RNA Longo não Codificante/isolamento & purificação
12.
Life Sci ; 231: 116571, 2019 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-31207308

RESUMO

AIMS: The role of long non-coding RNA's (lncRNA) in the biology of ulcerative colitis (UC) is not well understood. We have previously detected changes in lncRNA's associated with UC. This study aims to characterize one specific lncRNA, CDKN2B-AS1 whose expression was downregulated in UC patients. MAIN METHODS: UC biopsies were used to determine the levels of linear and circular CDKN2B-AS1 relative to healthy controls. In situ hybridization was used to determine the localization of CKDN2B-AS1 in the colon. The intestinal epithelial cell line, Caco-2, was used to study the effects of shRNA mediated loss of CDKN2B-AS1. Transepithelial electrical resistance was used to measure barrier function. An RT-PCR array, immunoblots and immunohistochemistry were used to determine tight junction proteins that CDKN2B-AS1 regulates. KEY FINDINGS: CDKN2B-AS1 is transcribed into not only linear transcripts but also as circular RNA through back-splicing and both forms are decreased in IBD. CDKN2B-AS1 is expressed mainly in colonic epithelial cells. Cells with down-regulated CDKN2B-AS1 exhibited increased proliferation and no alterations in apoptosis. Targeting both the linear and circular transcripts of CDKN2B-AS1 with short hairpin RNAs enhanced barrier function. We subsequently determined that Claudin-2, a "leaky Claudin" known to decrease barrier function, was decreased in CDKN2B-AS1 knockdown cells. SIGNIFICANCE: This study identifies a novel lncRNA with both linear and circular transcripts affecting UC biology.


Assuntos
Doenças Inflamatórias Intestinais/genética , RNA Longo não Codificante/biossíntese , RNA Longo não Codificante/genética , Adulto , Apoptose/genética , Células CACO-2 , Proliferação de Células/genética , Claudina-2/genética , Claudina-2/metabolismo , Colite Ulcerativa/genética , Colite Ulcerativa/metabolismo , Inibidor de Quinase Dependente de Ciclina p15/biossíntese , Inibidor de Quinase Dependente de Ciclina p15/genética , Inibidor de Quinase Dependente de Ciclina p15/metabolismo , DNA Circular/genética , Células Epiteliais/metabolismo , Feminino , Humanos , Doenças Inflamatórias Intestinais/patologia , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patologia , Masculino , RNA/genética , RNA Longo não Codificante/metabolismo
13.
Med Sci Monit ; 25: 3221-3230, 2019 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-31042695

RESUMO

BACKGROUND Recent studies have demonstrated that Linc00152 is highly expressed in multiple cancer types and its genes show tumor-promoting characteristics. However, the efficacy and biological mechanism of Linc00152 in bladder cancer remains unclear. MATERIAL AND METHODS We study investigated the relative expression and promoter methylation of Linc00152 in 126 cases of bladder cancer tissues by qRT-PCR and Bisulfite sequencing PCR. qRT-PCR was used to assess the relative expression of Linc00152 in 4 human bladder cancer cell lines. To explore the biological properties of Linc00152, we performed cell growth and soft-agar colony-formation assays, flow cytometry analyses, wound-healing assay, and Transwell assay. Western blot analysis was used to detect the underlying mechanisms of Linc00152 in bladder cancer. RESULTS We found that Linc00152 was highly expressed in 126 cases of bladder carcinoma tissues (p<0.001) and 4 cell lines (p<0.01), and Linc00152 is more commonly expressed in patients with advanced-stage cancer (p=0.021). Knockdown of Linc00152 by using siRNAs in bladder cancer cell lines (T24 and HT-1197) suppressed cell viability and growth by causing cell cycle arrest and apoptosis (p<0.001), as well as inhibiting cell migration and invasion (p<0.001). In addition, the quantitative RT-PCR and Western blot results suggest that knockdown of Linc00152 reduced Wnt/ß-Catenin signaling (p<0.001). CONCLUSIONS This research shows that Linc00152 is highly expressed in patients with bladder cancer and the possible carcinogenic effect of Linc00152 in bladder cancer occurs through activating the Wnt/ß-Catenin signaling pathway, and could be a new biomarker for diagnosis and prevention of this cancer.


Assuntos
RNA Longo não Codificante/biossíntese , Neoplasias da Bexiga Urinária/genética , Neoplasias da Bexiga Urinária/metabolismo , Via de Sinalização Wnt , Adulto , Idoso , Apoptose/fisiologia , Ciclo Celular/fisiologia , Pontos de Checagem do Ciclo Celular/fisiologia , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/fisiologia , Feminino , Técnicas de Silenciamento de Genes , Humanos , Masculino , Pessoa de Meia-Idade , Metástase Neoplásica , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Neoplasias da Bexiga Urinária/patologia
14.
Cell Biochem Funct ; 37(5): 348-358, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31066476

RESUMO

Pneumonia is an inflammatory disease that occurs in the lungs associated with pathogens or other factors. It has been well established that long noncoding RNA X inactivate-specific transcript (XIST) is involved in several cancers. The present study focused on the effect and detailed mechanism of XIST in lipopolysaccharide (LPS)-induced injury in pneumonia. Here, XIST was silenced by transfection with XIST-targeted siRNA, and then, mRNA expression, cell viability, apoptosis, and protein expression were, respectively, assessed by qRT-PCR, CCK-8, flow cytometry, and Western blotting. Luciferase reporter, RIP, and RNA pull-down assays were used to detect the combination of miR-370-3p and XIST. Besides, the tested proinflammatory factors were analysed by qRT-PCR and Western blot, and their productions were quantified by ELISA. The results showed that XIST expression was robustly increased in serum of patients with acute-stage pneumonia and LPS-induced WI-38 human lung fibroblasts cells. Functional analyses demonstrated that knockdown of XIST remarkably alleviated LPS-induced cell injury through increasing cell viability and inhibiting apoptosis and inflammatory cytokine levels. Mechanistically, XIST functioned as a competitive endogenous RNA (ceRNA) by effectively binding to miR-370-3p and then restoring TLR4 expression. More importantly, miR-370-3p inhibitor abolished the function of XIST knockdown on cell injury and JAK/STAT and NF-κB pathways. Taken together, XIST may be involved in progression of cell inflammatory response, and XIST/miR-370-3p/TLR4 axis thus may shed light on the development of novel therapeutics to the treatment of acute stage of pneumonia. SIGNIFICANCE OF THE STUDY: Our study demonstrated that XIST was highly expressed in patients with acute stage of pneumonia. Knockdown of XIST remarkably alleviated LPS-induced cell injury through increasing cell viability and inhibiting apoptosis and inflammatory cytokine levels through regulating JAK/STAT and NF-κB pathways.


Assuntos
Apoptose/efeitos dos fármacos , Inflamação/tratamento farmacológico , Lipopolissacarídeos/farmacologia , MicroRNAs/antagonistas & inibidores , Pneumonia/tratamento farmacológico , RNA Longo não Codificante/genética , RNA Interferente Pequeno/farmacologia , Receptor 4 Toll-Like/antagonistas & inibidores , Doença Aguda , Adulto , Células Cultivadas , Feminino , Humanos , Inflamação/induzido quimicamente , Inflamação/metabolismo , Lipopolissacarídeos/antagonistas & inibidores , Masculino , MicroRNAs/análise , MicroRNAs/metabolismo , NF-kappa B/metabolismo , Pneumonia/diagnóstico , Pneumonia/metabolismo , RNA Longo não Codificante/biossíntese , Transdução de Sinais/efeitos dos fármacos , Receptor 4 Toll-Like/análise , Receptor 4 Toll-Like/metabolismo
15.
Biomed Res Int ; 2019: 1275491, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31061820

RESUMO

Background: As a newly discovered lncRNA, bladder cancer-associated transcript 1 (BLACAT1) has been reported to correlate with poor clinical outcomes in several different cancers. This study aimed to evaluate its generalized predictive value for cancer prognosis. Materials and Methods: We thoroughly searched PubMed, Embase, and Web of Science databases for eligible studies published until November 11, 2018, in which the relationship between BLACAT1 expression and cancer prognosis was explored. The analyses were performed using Review Manager Version 5.3 and Stata SE 12.0. The primary endpoints included overall survival (OS), pathological characteristics (TNM stage and tumor grade), lymph node metastasis (LNM), and distant metastasis. Results: Ten studies containing 861 patients with 7 different cancerous diseases were eventually included. The results demonstrated that patients with high lncRNA BLACAT1 expression had a significantly shorter OS (HR: 1.82, 95% CI: 1.44-2.30, p < 0.00001) than patients with low lncRNA BLACAT1 expression. Moreover, elevated BLACAT1 expression was significantly associated with advanced TNM stage (OR: 2.29, 95% CI: 1.15-4.56, p = 0.005), high tumor grade (OR: 1.67, 95% CI: 1.11-2.53, p = 0.01), and lymph node metastasis (OR: 2.53, 95% CI: 1.80-3.57, p < 0.00001). Meanwhile, the expression of BLACAT1 had no significant association with age (p = 0.92), gender (p = 0.55), and smoking (p = 0.62). Conclusion: High expression of lncRNA BLACAT1 may predict a poor prognosis in OS, TNM stage, tumor grade, and LNM. Its predictive roles were not significantly affected by age, gender, or smoking. Therefore, lncRNA BLACAT1 may serve as a promising predictor in cancer prognosis.


Assuntos
Biomarcadores Tumorais/biossíntese , Regulação Neoplásica da Expressão Gênica , Neoplasias/metabolismo , Neoplasias/mortalidade , RNA Longo não Codificante/biossíntese , Intervalo Livre de Doença , Feminino , Humanos , Metástase Linfática , Masculino , Estadiamento de Neoplasias , Neoplasias/patologia , Valor Preditivo dos Testes , RNA Neoplásico , Taxa de Sobrevida
16.
Biomed Res Int ; 2019: 5437531, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30949502

RESUMO

Glioma is a lethal, malignant intracranial tumor that becomes progressively common. It has been shown that long noncoding RNAs (lncRNAs) serve important roles in numerous diseases such as gliomas. lncRNAs can regulate the expression of targeted genes through various mechanisms. To identify a novel lncRNA that may be critical in glioma, the present study downloaded the RNA expression profiles of 171 glioma tissues and 5 normal tissues from The Cancer Genome Atlas (TCGA) database using the TCGAbiolinks package in R. Then, lncRNAs in the downloaded TCGA data were identified using the HUGO Gene Nomenclature Committee (HGNC). Based on the fragments per kilobase million value, differential expression analysis was conducted using the limma package in R. In addition, receiver operating characteristic (ROC) analysis was performed, and the area under the curve (AUC) was evaluated using the ROCR package in R. A total of 178 lncRNAs corresponding to differentially expressed genes with an AUC >0.85 were selected. Upon identifying the differential lncRNAs, ceRNA networks were constructed with these differential lncRNAs using the starbase database. From these networks, the top 10% hub genes were selected. In addition, the present study randomly selected 4 lncRNAs for quantitative polymerase chain reaction validation in tissue samples. The results revealed that lncRNA ASB16-AS1 exhibited significantly differential expression in tissue samples and was significantly associated with tumor staging and grading. Furthermore, the proliferation, invasion, and migration of U87MG and U251 glioblastoma stem-like cells (U87GS, U251GS) were significantly inhibited upon inhibition of ASB16-AS1, and the expression of key proteins in the EMT signaling pathway was affected by knocking down ASB16-AS1. Overall, the present study revealed that lncRNA ASB16-AS1 improves the proliferation, migration, and invasion of glioma cells.


Assuntos
Neoplasias Encefálicas/metabolismo , Movimento Celular , Proliferação de Células , Regulação Neoplásica da Expressão Gênica , Glioma/metabolismo , RNA Longo não Codificante/biossíntese , RNA Neoplásico/biossíntese , Adulto , Idoso , Idoso de 80 Anos ou mais , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/patologia , Linhagem Celular Tumoral , Feminino , Glioma/genética , Glioma/patologia , Humanos , Masculino , Pessoa de Meia-Idade , Invasividade Neoplásica , RNA Longo não Codificante/genética , RNA Neoplásico/genética , Transdução de Sinais/genética
17.
Medicine (Baltimore) ; 98(17): e15251, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31027073

RESUMO

BACKGROUND: Aberrant expression of long non-coding RNA Zinc finger E-box binding homeobox 1 antisense 1 (lncRNA ZEB1-AS1) can be detected in numerous malignancies. Therefore, a meta-analysis had been carried out in this study, aiming to examine the prognostic value of lncRNA ZEB1-AS1 in malignancies. METHODS: Electronic databases, such as PubMed, OVID as well as Web of Science, had been systemically retrieved from inception to February 14th, 2019. Besides, the hazard ratios (HRs), together with the corresponding 95% confidence intervals (CIs), had also been analyzed for determining the association of lncRNA ZEB1-AS1 expression with the overall survival (OS) and recurrence-free survival (RFS). In addition, the pooled odds ratios (ORs) would also be computed using the Stata SE12.0 software for evaluating the relationship of lncRNA ZEB1-AS1 expression with pathological factors. RESULTS: A total of 21 original studies involving 1801 cancer patients had been enrolled into the current meta-analysis. As suggested by the pooled HR, high expression of lncRNA ZEB1-AS1 had displayed marked correlation with OS (HR = 2.16, 95% CI: 1.89-2.47) among cancer patients, and no significant heterogeneity was detected. Additionally, high expression of lncRNA ZEB1-AS1 was also markedly associated with RFS among cancer patients (pooled HR = 2.55, 95% CI: 1.61-4.03). Besides, the expression of lncRNA ZEB1-AS1 had displayed marked correlation with poor histological grade (PHG) (OR = 2.86, 95% CI: 2.11-3.87), high tumor stage (HTS) (OR = 3.81, 95% CI: 2.72-5.34) as well as lymph node metastasis (LNM) (OR = 3.33, 95% CI: 2.47-4.49). Additionally, no distinct asymmetry had been detected for RFS, PHG as well as HTS based on Begg funnel plot. CONCLUSIONS: Taken together, high expression of lncRNA ZEB1-AS1 can predict the dismal OS, RFS, LNM, PHG, and HTS, indicating that lncRNA ZEB1-AS1 can be potentially used as a new biomarker to predict the dismal prognosis for cancer patients.


Assuntos
Neoplasias/metabolismo , Neoplasias/mortalidade , RNA Longo não Codificante/biossíntese , Biomarcadores Tumorais , China , Estudos Clínicos como Assunto , Humanos , Metástase Linfática , Recidiva Local de Neoplasia , Estadiamento de Neoplasias , Razão de Chances , Prognóstico , Análise de Sobrevida
18.
Biomed Res Int ; 2019: 3526407, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31019967

RESUMO

Background: Cholangiocarcinoma (CCA) is the second most common malignant primary liver tumor and has shown an alarming increase in incidence over the last two decades. However, the mechanisms behind tumorigenesis and progression remain insufficient. The present study aimed to uncover the underlying regulatory mechanism on CCA and find novel biomarkers for the disease prognosis. Method: The RNA-sequencing (RNA-seq) datasets of lncRNAs, miRNAs, and mRNAs in CCA as well as relevant clinical information were obtained from the Cancer Genome Atlas (TCGA) database. After pretreatment, differentially expressed RNAs (DERNAs) were identified and further interrogated for their correlations with clinical information. Prognostic RNAs were selected using univariate Cox regression. Then, a ceRNA network was constructed based on these RNAs. Results: We identified a total of five prognostic DEmiRNAs, 63 DElncRNAs, and 90 DEmRNAs between CCA and matched normal tissues. Integrating the relationship between the different types of RNAs, an lncRNA-miRNA-mRNA network was established and included 28 molecules and 47 interactions. Screened prognostic RNAs involved in the ceRNA network included 3 miRNAs (hsa-mir-1295b, hsa-mir-33b, and hsa-mir-6715a), 7 lncRNAs (ENSG00000271133, ENSG00000233834, ENSG00000276791, ENSG00000241155, COL18A1-AS1, ENSG00000274737, and ENSG00000235052), and 18 mRNAs (ANO9, FUT4, MLLT3, ABCA3, FSCN2, GRID2IP, NCK2, MACC1, SLC35E4, ST14, SH2D3A, MOB3B, ACTL10, RAB36, ATP1B3, MST1R, SEMA6A, and SEL1L3). Conclusions: Our study identified novel prognostic makers and predicted a previously unknown ceRNA regulatory network in CCA and may provide novel insight into a further understanding of lncRNA-mediated ceRNA regulatory mechanisms in CCA.


Assuntos
Neoplasias dos Ductos Biliares , Colangiocarcinoma , Neoplasias Hepáticas , MicroRNAs , RNA Longo não Codificante , RNA Mensageiro , RNA Neoplásico , Idoso , Neoplasias dos Ductos Biliares/genética , Neoplasias dos Ductos Biliares/metabolismo , Neoplasias dos Ductos Biliares/patologia , Colangiocarcinoma/genética , Colangiocarcinoma/metabolismo , Colangiocarcinoma/mortalidade , Intervalo Livre de Doença , Feminino , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/mortalidade , Masculino , MicroRNAs/biossíntese , MicroRNAs/genética , Pessoa de Meia-Idade , RNA Longo não Codificante/biossíntese , RNA Longo não Codificante/genética , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , RNA Neoplásico/biossíntese , RNA Neoplásico/genética , Análise de Sequência de RNA , Taxa de Sobrevida
19.
Med Sci Monit ; 25: 2845-2851, 2019 Apr 18.
Artigo em Inglês | MEDLINE | ID: mdl-30996220

RESUMO

BACKGROUND lncRNA BANCR participates in the pathogenesis of various types of human diseases; however, its involvement in diabetic retinopathy is unknown. MATERIAL AND METHODS In this study, the expression of lncRNA BANCR in plasma of patients with diabetic retinopathy, diabetic patients without complications, and healthy controls was analyzed by qRT-PCR. The accuracy plasma BANCR in diagnosing diabetic retinopathy was analyzed by ROC curve analysis. lncRNA BANCR expression vector and siRNA were transfected into the human retinal pigment epithelial cell line ARPE-19, and cell apoptosis was analyzed by cell apoptosis assay. RESULTS We found that lncRNA BANCR was significantly upregulated in patients with diabetic retinopathy compared to diabetic patients without complications and healthy controls. Upregulation of lncRNA BANCR effectively distinguished patients with diabetic retinopathy from diabetic patients without complications and healthy controls. High-glucose treatment led to upregulated expression of BANCR in the human retinal pigment epithelial cell line ARPE-19. Overexpression of BANCR promoted and siRNA silencing inhibited the apoptosis of cells in the human retinal pigment epithelial cell line ARPE-19 in a high-glucose environment. CONCLUSIONS lncRNA BANCR is overexpressed in patients with diabetic retinopathy and can promote apoptosis of retinal pigment epithelial cells.


Assuntos
Retinopatia Diabética/genética , RNA Longo não Codificante/biossíntese , RNA Longo não Codificante/genética , Epitélio Pigmentado da Retina/metabolismo , Adulto , Idoso , Apoptose/fisiologia , Estudos de Casos e Controles , Linhagem Celular , Proliferação de Células/fisiologia , Retinopatia Diabética/sangue , Retinopatia Diabética/metabolismo , Retinopatia Diabética/patologia , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Feminino , Glucose/metabolismo , Humanos , Masculino , Pessoa de Meia-Idade , RNA Longo não Codificante/sangue , Epitélio Pigmentado da Retina/patologia , Pigmentos da Retina/metabolismo , Ativação Transcricional , Regulação para Cima
20.
Acta Ophthalmol ; 97(6): e902-e912, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30900812

RESUMO

PURPOSE: To evaluate the effect of conbercept on the expression of long noncoding RNAs (lncRNAs) and mRNAs in the fibrovascular membranes of proliferative diabetic retinopathy (PDR) patients. METHODS: Twenty patients, diagnosed with PDR, who underwent pars plana vitrectomy (PPV), were recruited for this study. Ten patients were treated for PPV alone (Control Group), and the others received conbercept injections before PPV (Treated Group). The fibrovascular membranes were harvested during surgery. Expression of lncRNAs and mRNAs in the membranes was tested using lncRNA Arrays. Bioinformatics analyses were performed to identify the related biological modules and pathways of the differentially expressed genes. A lncRNA/mRNA coexpression network was built to identify the correlations between lncRNAs and mRNAs. Real-time PCR was conducted to verify the microarray results. RESULTS: We identified 427 differentially expressed lncRNAs, of which 263 were upregulated and 164 were downregulated. Gene ontology (GO) analysis indicated that these lncRNAs-coexpressed mRNAs targeted various metabolic processes, especially the gluconeogenesis. Kyoto Encyclopaedia of Genes and Genomes (KEGG) results indicated that 16 pathways had significant differences in gene expression, including gluconeogenesis, HIF-1 signalling pathway, NOD-like receptor pathway, etc. The lncRNA/mRNA coexpression network revealed that many differentially expressed lncRNAs were enriched in the HIF-1, TNF-α and NOD-like receptor pathways. LincRNAs were the largest category and further bioinformatics analysis implied that these lincRNAs-coexpressed mRNAs were mainly involved in PDR-related biological processes and pathological pathways. CONCLUSION: Conbercept treatment can change the expression profiles of lncRNAs and mRNAs in the fibrovascular membranes of PDR patients. A complete understanding of the relationship between lncRNAs and anti-VEGF drugs may contribute to new therapeutic regimen for PDR.


Assuntos
Retinopatia Diabética/tratamento farmacológico , Regulação da Expressão Gênica/efeitos dos fármacos , RNA Longo não Codificante/genética , RNA Mensageiro/genética , Proteínas Recombinantes de Fusão/administração & dosagem , Idoso , Retinopatia Diabética/genética , Retinopatia Diabética/metabolismo , Feminino , Perfilação da Expressão Gênica , Humanos , Injeções Intravítreas , Masculino , Pessoa de Meia-Idade , RNA Longo não Codificante/biossíntese , Reação em Cadeia da Polimerase em Tempo Real , Transdução de Sinais
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