RESUMO
Circular RNAs (circRNAs) play functional roles in rheumatoid arthritis (RA) progression. Fibroblast-like synoviocytes (RASFs) are the main effectors in RA development. In this study, we explored the function and mechanism of circ_0008410 in RASFs. qRT-PCR was used to detect the expression of circ_0008410, microRNA-149-5p (miR-149-5p), and homeodomain-interacting protein kinase 2 (HIPK2). Cell counting kit-8, EdU assay, flow cytometry, and transwell assay were performed to evaluate cell proliferation, apoptosis, migration, and invasion. Western blot measured the protein levels of related markers and HIPK2. The levels of IL-1ß, TNF-α, and IL-6 were tested by corresponding ELISA kits and Western blot. The combination between miR-149-5p and circ_0008410 or HIPK2 was detected by dual-luciferase reporter assay or RNA immunoprecipitation (RIP) assay. Our data showed that circ_0008410 and HIPK2 were elevated, while miR-149-5p was downregulated in RA synovial tissues and RASFs. Circ_0008410 promoted RASF proliferation, migration, invasion, and inflammation while inhibiting apoptosis. MiR-149-5p was a target of circ_0008410, and its overexpression could reverse the promoting effects of circ_0008410 on RASF dysfunction. Moreover, miR-149-5p could target HIPK2 to suppress RASF proliferation, migration, invasion, and inflammation. Collectively, circ_0008410 promoted RASF dysfunction via miR-149-5p/HIPK2, which might provide a potential target for RA therapy.
Assuntos
Artrite Reumatoide , MicroRNAs , Sinoviócitos , Humanos , Membrana Sinovial , Apoptose/genética , Artrite Reumatoide/genética , Proliferação de Células , Fibroblastos , Inflamação , MicroRNAs/genética , Proteínas de Transporte , Proteínas Serina-Treonina Quinases/genéticaRESUMO
5-Fluorouracil is a commonly used chemotherapy drug for colorectal cancer. Resistance to 5-Fluorouracil remains a challenge. This research aimed to explore the mechanism of 5-Fluorouracil resistance in colorectal cancer. RT-qPCR and Western blot were used to determine the RNA and protein expression in both cells and exosome. Assays in vitro and in vivo were performed to measure the role of miR-149-5p in colorectal cancer cells. RIP, luciferase activity report, and RNA pulldown assay were applied to detect the association of PTOV1-AS1, SUV39H1, miR-149-5p, and FOXM1. MiR-149-5p was down-expressed in 5-Fluorouracil-resistant cells. MiR-149-5p enhanced the effectiveness of 5-Fluorouracil both in vitro and in vivo. Sensitive colorectal cancer cells released exosomal miR-149-5p to sensitize resistant cells to chemotherapy. Mechanistically, miR-149-5p targeted the FOXM1 to inactivate Wnt/ß-catenin pathway, and PTOV1-AS1 recruited SUV39H1 to suppress miR-149-5p transcription, in turn activating Wnt/ß-catenin pathway, and forming a positive feedback loop with FOXM1. PTOV1-AS1 inhibits miR-149-5p by a positive feedback loop with FOXM1-mediated Wnt/ß-catenin pathway, which provides insights into a potential novel target for enhancing the effectiveness of chemotherapy in colorectal cancer patients.
Assuntos
Neoplasias Colorretais , MicroRNAs , RNA Longo não Codificante , Humanos , MicroRNAs/genética , beta Catenina/metabolismo , Linhagem Celular Tumoral , Retroalimentação , Proliferação de Células , Via de Sinalização Wnt , Fluoruracila , Neoplasias Colorretais/genética , Regulação Neoplásica da Expressão Gênica , RNA Longo não Codificante/genética , Proteínas de Neoplasias/metabolismo , Biomarcadores Tumorais/uso terapêuticoRESUMO
As one of the important traits in pig production, meat quality has important research significance and value. Intramuscular fat (IMF) content is one of the most important factors affecting pork quality. Many experimental studies have shown that IMF content is closely related to the flavor, tenderness, and juiciness of pork. Therefore, it is of great significance to study the mechanism of porcine IMF deposition. Previous research indicated that miR-149-5p promoted the proliferation of porcine intramuscular (IM) preadipocytes and decreased their ability to differentiate, albeit the exact mechanism of action is unknown. In vitro, foreign pigs showed increased miR-149-5p expression and reduced fat deposition when compared to Queshan Black pigs. This study conducted metabolomics and transcriptomics analyses of porcine IM preadipocytes overexpressing miR-149-5p to verify their effects on lipid formation. According to metabolomics analysis, the overexpression of miR-149-5p has significantly altered the lipid, organic acid, and organic oxygen metabolites of porcine IM preadipocytes. Specially speaking, it has changed 115 metabolites, including 105 up-regulated and 10 down-regulated ones, as well as the composition of lipid, organic acid, and organic oxygen metabolism-related metabolites. RNA-seq analysis showed that overexpression of miR-149-5p significantly altered 857 genes, of which 442 were up-regulated, and 415 were down-regulated, with enrichment to MAPK, IL-17, PI3K-Akt, and ErbB signaling pathways. We found that overexpression of miR-149-5p inhibited adipogenic differentiation by changing cAMP signaling pathway in porcine IM preadipocytes. In addition, the overexpression of miR-149-5p may affect the transport of Cu2+ by targeting ATP7A and inhibiting adipogenic differentiation. These findings elucidate the regulatory function of miR-149-5p in porcine IM preadipocytes, which may be a key target for controlling pork quality.
Assuntos
Adipócitos , MicroRNAs , Suínos , Animais , Adipócitos/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Transcriptoma , Fosfatidilinositol 3-Quinases/metabolismo , Adipogenia/genética , Lipídeos , Diferenciação Celular/genéticaRESUMO
BACKGROUND: Psoriasis is a chronic autoimmune skin disease with unclear pathogenesis. It was found that miR-149-5p was significantly decreased in psoriatic lesion tissues. In this study, we aims to investigate the role and related molecular mechanism of miR-149-5p on psoriasis. METHOD: IL-22 was used to stimulate HaCaT and NHEK cells to establish psoriasis model in vitro. The miR-149-5p and phosphodiesterase 4D (PDE4D) expression levels were detected by quantitative real-time PCR. HaCaT and NHEK cells proliferation was determined by Cell Couting Kit-8 assay. The cell apoptosis and cell cycle were detected by flow cytometry. The cleaved Caspase-3, Bax and Bcl-2 protein expressions were detected by western blot. The targeting relationship between PDE4D and miR-149-5p was predicted and confirmed by Starbase V2.0 and dual-luciferase reporter assay, respectively. RESULT: There was a low expression level of miR-149-5p and a high expression of PDE4D in psoriatic lesion tissues. MiR-149-5p could target PDE4D. IL-22 promoted HaCaT and NHEK cells proliferation, while inhibited cell apoptosis and accelerated cell cycle. Moreover, IL-22 decreased the expressions of cleaved Caspase-3 and Bax, and increased the expression of Bcl-2. And the overexpressed miR-149-5p promoted HaCaT and NHEK cells apoptosis, inhibited cell proliferation and retarded cell cycle, meanwhile increased the cleaved Caspase-3 and Bax expressions, decreased the Bcl-2 expression. In addition, PDE4D overexpression has the opposite effect as miR-149-5p. CONCLUSION: The overexpressed miR-149-5p inhibits IL-22-stimulated HaCaT and NHEK keratinocytes proliferation, promotes cell apoptosis and retards cell cycle by down-regulating the expression of PDE4D, which could be the promising therapeutic target of psoriasis.
Assuntos
MicroRNAs , Psoríase , Humanos , Apoptose/genética , Proteína X Associada a bcl-2/metabolismo , Proteína X Associada a bcl-2/uso terapêutico , Caspase 3/genética , Caspase 3/metabolismo , Ciclo Celular/genética , Proliferação de Células/genética , Nucleotídeo Cíclico Fosfodiesterase do Tipo 4/genética , Nucleotídeo Cíclico Fosfodiesterase do Tipo 4/metabolismo , Queratinócitos/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Psoríase/metabolismo , Interleucina 22RESUMO
Circular RNAs participate in the pathogenesis of various tumors, including gastric cancer (GC). In this study, we investigated the role of circBGN in regulating proliferation and invasion of GC cells and elucidated the mechanism. The expression of circBGN was assessed by quantitative reverse-transcription PCR and in situ hybridization. In addition, loss- and gain-of-function investigations in vitro and in vivo were performed to determine the biological functions of circBGN. Luciferase reporter assays and rescue experiments were applied to investigate the interaction between circBGN and miR-149-5p as well as the relationship between miR-149-5p and IL6. Our results showed that circBGN expression was significantly elevated in GC tissues and cells. Knockdown of circBGN dramatically suppressed GC cell proliferation and invasion in vitro. Xenograft experiments revealed that knockdown of circBGN delayed tumor growth in vivo. Furthermore, circBGN can directly bind to miR-149-5p, thereby preventing miR-149-5p from binding to its target mRNA [IL6 mRNA], thus activating IL6/STAT3 signaling pathway. Rescue assays indicated that circBGN regulates GC cell proliferation and invasion by upregulating miR-149-5p/IL6 axis output. Taken together, our investigation indicates that circBGN supports GC progression by activating IL6/STAT3 signaling pathway, thus pointing to a new possible therapeutic target in GC.
Assuntos
MicroRNAs , Neoplasias Gástricas , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células/genética , Regulação Neoplásica da Expressão Gênica , Humanos , Interleucina-6/genética , Interleucina-6/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , RNA Circular/genética , RNA Mensageiro , Fator de Transcrição STAT3/genética , Fator de Transcrição STAT3/metabolismo , Transdução de Sinais/genética , Neoplasias Gástricas/patologiaRESUMO
Epidemiological evidence has shown that fine particulate matter (PM2.5)-triggered inflammatory cascades are pivotal causes of chronic obstructive pulmonary disease (COPD). However, the specific molecular mechanism involved in PM2.5-induced COPD has not been clarified. Herein, we found that PM2.5 significantly downregulated miR-149-5p and activated the mitogen-activated protein kinase (MAPK) and nuclear factor-kappa B (NF-κB) signaling pathways and generated the inflammatory response in COPD mice and in human bronchial epithelial (BEAS-2B) cells. We determined that increased expression of interleukin-1ß (IL-1ß), IL-6, IL-8, and tumor necrosis factor-α (TNF-α) induced by PM2.5 was associated with decreased expression of miR-149-5p. The loss- and gain-of-function approach further confirmed that miR-149-5p could inhibit PM2.5-induced cell inflammation in BEAS-2B cells. The double luciferase reporter assay showed that miR-149-5p directly targeted TGF-beta-activated kinase 1 binding protein 2 (TAB2), which regulates the MAPK and NF-κB signaling pathways. We showed that miR-149-5p mediated the inflammatory response by targeting the 3'-UTR sequence of TAB2 and that it subsequently weakened the TAB2 promotor effect via the MAPK and NF-κB signaling pathways in BEAS-2B cells exposed to PM2.5. Thus, miR-149-5p may be a key factor in PM2.5-induced COPD. This study improves our understanding of the molecular mechanism of COPD.
Assuntos
MicroRNAs , Doença Pulmonar Obstrutiva Crônica , Humanos , Camundongos , Animais , NF-kappa B/metabolismo , Proteínas Quinases Ativadas por Mitógeno/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Transdução de Sinais , Material Particulado/toxicidade , Inflamação/genética , Inflamação/patologia , Doença Pulmonar Obstrutiva Crônica/genética , Doença Pulmonar Obstrutiva Crônica/metabolismo , Proteínas Adaptadoras de Transdução de Sinal/genética , Proteínas Adaptadoras de Transdução de Sinal/metabolismoRESUMO
BACKGROUND: Psoriasis is a chronic, complicated, and recurrent inflammatory skin disease, whose precise molecular mechanisms need to be further explored. The lncRNA bladder cancer-associated transcript 1 (BLACAT1) is aberrantly expressed in many cancers and associated with cellular hyperproliferation and may play a role in the pathogenesis of psoriasis. Thus, this study aimed at identifying the primary mechanism associated with BLACAT1 in psoriasis pathogenesis. MATERIALS AND METHODS: Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) was performed to detect the expression of BLACAT1 in psoriasis tissues. Cell proliferation and apoptosis were assessed using cell counting kit-8 and apoptosis assays, respectively. In vivo experiments and histopathological examinations were performed to investigate the effects of BLACAT1 on psoriasis. Dual-luciferase Reporter and RNA immunoprecipitation assays were used to evaluate the relationship among BLACAT1 and miR-149-5p and AKT1. RESULTS: BLACAT1 was upregulated in psoriasis tissues. Overexpression exacerbated the clinical manifestation of psoriasis and increased the epidermal thickness in imiquimod-induced mice. BLACAT1 could promote proliferation and inhibit apoptosis of keratinocytes. Further studies demonstrated that BLACAT1 positively regulated AKT1 expression, functioning as a competing endogenous RNA (ceRNA) by sponging miR-149-5p. CONCLUSIONS: The combination of lncRNA BLACAT1 and miR-149-5p regulates AKT1 expression and promotes psoriasis formation thus may provide a new direction for psoriasis treatment.
Assuntos
MicroRNAs , Psoríase , RNA Longo não Codificante , Neoplasias da Bexiga Urinária , Animais , Camundongos , MicroRNAs/genética , MicroRNAs/metabolismo , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Psoríase/genética , Queratinócitos/metabolismo , Apoptose/genética , Proliferação de CélulasRESUMO
Abundant regulatory genes and complex circuits involving non-coding RNAs (ncRNAs) monitor the formation and development of hepatic fibrosis (HF). Circular RNAs (circRNAs) are a class of RNAs generated from protein coding genes by back-splicing, playing crucial roles in various pathological processes, including HF. However, little is known about mechanisms of action of circRNAs, let alone in HF. In this study, we found circUbe2k enhanced in CCl4 -induced HF mice and LX-2 cells stimulated with TGF-ß1, regulating the development of HF. Restraining the expression of circUbe2k inhibited α-SMA and Col1α1 expression in CCl4 -induced HF mice and in LX-2 cells stimulated with TGF-ß1. Furthermore, inhibiting circUbe2k expression reduced hepatic stellate cells (HSCs) activation and proliferation in vivo and in vitro. Mechanistically, we demonstrated a direct interaction between circUbe2k and miR-149-5p, which results in the modulation of TGF-ß2 expressions. Together, circUbe2k may act as a "catalyst" of HSCs activation and HF through the circUbe2k/miR-149-5p/TGF-ß2 axis. Our results provide unprecedented evidence for a significant role for circUbe2k to serve as a potential biomarker for HF therapy.
Assuntos
Regulação da Expressão Gênica , Cirrose Hepática/patologia , MicroRNAs/genética , RNA Circular/genética , Fator de Crescimento Transformador beta2/metabolismo , Enzimas de Conjugação de Ubiquitina/genética , Animais , Tetracloreto de Carbono , Proliferação de Células , Cirrose Hepática/induzido quimicamente , Cirrose Hepática/genética , Cirrose Hepática/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Transdução de Sinais , Fator de Crescimento Transformador beta2/genéticaRESUMO
BACKGROUND: The initiation and development of thyroid cancer may be associated with the deregulation of circular RNAs (circRNAs). The purpose of this work was to explore the role of circRNA casein kinase 1 gamma 1 (circ-CSNK1G1) in thyroid cancer. METHODS: The expression of circ-CSNK1G1, miR-149-5p, and mitogen-activated protein kinase 1 (MAPK1) was concluded using quantitative real-time PCR (qPCR), and the expression of MAPK1 protein was detected by Western blot assay. Cell viability was monitored by CCK-8 assay. Cell proliferation was determined by colony formation assay and EdU assay. Cell apoptosis and cycle were checked by flow cytometry assay. Cell invasion was determined by transwell assay. The predicted binding relationship between miR-149-5p and circ-CSNK1G1 or MAPK1 was verified by dual-luciferase reporter assay. The role of circ-CSNK1G1 in vivo was determined by establishing animal models. RESULTS: The present work discovered the upregulation of circ-CSNK1G1 in tumor tissues of thyroid cancer. In function, circ-CSNK1G1 knockdown inhibited proliferation, survival, and invasion in cancer cells, and tumor growth in mouse models. MiR-149-5p was a target of circ-CSNK1G1, and the anti-tumor effects of circ-CSNK1G1 knockdown were abolished by miR-149-5p downregulation. In addition, miR-149-5p directly targeted MAPK1, and miR-149-5p restoration-inhibited cell proliferation and invasion were recovered by MAPK1 overexpression. CONCLUSION: Circ-CSNK1G1 acted as miR-149-5p to relieve the inhibition of miR-149-5p on MAPK1, thus promoting the malignant development of thyroid cancer.
Assuntos
Caseína Quinase I/metabolismo , MicroRNAs/metabolismo , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Neoplasias da Glândula Tireoide/metabolismo , Adulto , Animais , Linhagem Celular Tumoral , Modelos Animais de Doenças , Regulação para Baixo , Feminino , Humanos , Masculino , Camundongos , Camundongos Nus , Pessoa de Meia-Idade , Neoplasias da Glândula Tireoide/genéticaRESUMO
Earlier studies have shown that circular RNA (circRNA) expression is closely related to the malignant progression of cancer, but the role of circ-DONSON in gastric cancer (GC) has not been fully elucidated. The expression of circ-DONSON, miR-149-5p and lactate dehydrogenase A (LDHA) was measured via qRT-PCR. CCK8 assay was used to assess cell viability, and colony formation assay was performed to detect the number of colonies and the radiosensitivity of cells. Besides, flow cytometry, transwell assay and tube formation assay were employed to determine cell apoptosis, migration, invasion and angiogenesis. Western blot analysis was used to assess the protein expression. The interaction between miR-149-5p and circ-DONSON or LDHA was confirmed by dual-luciferase reporter assay. The influence of circ-DONSON on GC tumor growth in vivo was explored through constructing mice xenograft models. Our results suggested that circ-DONSON was highly expressed in GC tissues and cells. Loss-functional assay results confirmed that silenced circ-DONSON could inhibit the proliferation, metastasis and angiogenesis, while enhance the apoptosis and radiosensitivity of GC cells. In terms of mechanism, circ-DONSON could sponge miR-149-5p, which could target LDHA in GC. MiR-149-5p inhibitor or LDHA overexpression could reverse the suppression effect of circ-DONSON knockdown on GC progression. Additionally, our results also suggested that circ-DONSON silencing could restrain the tumor growth of GC in vivo. These results demonstrated that circ-DONSON could facilitate GC progression by increasing LDHA expression via sponging miR-149-5p, indicating that circ-DONSON might be a novel biomarker for GC treatment.
Assuntos
Lactato Desidrogenase 5 , MicroRNAs , RNA Circular , Neoplasias Gástricas , Animais , Proliferação de Células , Humanos , L-Lactato Desidrogenase , Lactato Desidrogenase 5/genética , Camundongos , MicroRNAs/genética , RNA Circular/genética , Neoplasias Gástricas/patologiaRESUMO
Triple-negative breast cancer (TNBC) is a breast cancer subtype without targeted treatment options. Accumulating evidence has demonstrated the roles of circular RNAs in cancer. This study aimed to investigate the expression and function of circFAM64A in TNBC. The GSE101124 dataset from the GEO database was examined to identify the differentially expressed circular RNAs in TNBC. RT-qPCR and western blot analyses were performed to measure gene expression. TNBC cell proliferation, migration, invasion, and cell cycle were assessed using cell counting kit-8, EdU, flow cytometry, wound healing, and transwell invasion experiments. Bioinformatics analysis, RIP, RNA pulldown, and luciferase reporter assays were used to investigate the regulatory mechanism of circFAM64A. In this study, CircFAM64A expression was significantly upregulated in TNBC tissues and cells compared with normal tissues and cells. Overexpression of circFAM64A increased the proliferative, migratory, and invasive capacities of TNBC cells and promoted cell cycle progression. Mechanistically, circFAM64A acted as a molecular sponge for miR-149-5p, and miR-149-5p directly targeted the Cdc10-dependent transcript 1 (CDT1) 3'UTR. Moreover, the high expression of CDT1 is associated with a poor prognosis in patients with breast cancer. Rescue experiments demonstrated that circFAM64A sponged miR-149-5p to increase CDT1 expression, thereby promoting cellular processes in TNBC. Overall, CircFAM64A plays an oncogenic role in TNBC by interacting with miR-149-5p to increase CDT1 expression.
Assuntos
Peptídeos e Proteínas de Sinalização Intracelular/genética , MicroRNAs , Proteínas Nucleares/genética , Neoplasias de Mama Triplo Negativas , Proteínas de Ciclo Celular/metabolismo , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Regulação Neoplásica da Expressão Gênica/genética , Humanos , MicroRNAs/genética , MicroRNAs/metabolismo , Neoplasias de Mama Triplo Negativas/genéticaRESUMO
Accumulating evidence indicates that microRNAs (miRs) play critical roles in essentially all biological processes and their altered expression has been documented in various disease conditions, including human malignancies. Although several cellular mechanisms have been identified in mediating the effects of miRs, the involvement of G-protein-coupled, platelet-activating factor-receptor (PAFR) signaling in miR-149-5p-induced effects on lung cancer growth and therapeutic potential has not been studied. To that end, we first evaluated the functional significance of PAFR and miR-149-5p in A549 and H1299 human non-small cell lung cancer (NSCLC) cell lines. We observed that these tumor lines express endogenous PAFR and miR-149-5p and that PAFR activation by PAF agonist (CPAF) significantly increased, whereas miR-149-5p mimic transfection inhibited cell proliferation in a dose-dependent manner. Interestingly, miR-149-5p mimic significantly attenuated CPAF-mediated increased proliferation of NSCLC cells, as confirmed by miR-149-5p, cyclin D1, and forkhead box protein M1 (FOXM1) expression analysis via qPCR. Our next studies examined PAFR- and miR-149-5p-mediated effects on targeted therapy (i.e., erlotinib and gefitinib) responses. We observed that erlotinib and gefitinib inhibited A549 and H1299 cell survival in a dose- and time-dependent manner, and CPAF significantly blocked this effect. These findings indicate that miR-149-5p blocks PAFR-mediated increased cell proliferation, and PAFR activation attenuates the cytotoxic effects of targeted therapy.
Assuntos
Fenômenos Biológicos , Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , MicroRNAs , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/patologia , Linhagem Celular Tumoral , Proliferação de Células , Cloridrato de Erlotinib/farmacologia , Gefitinibe/farmacologia , Gefitinibe/uso terapêutico , Regulação Neoplásica da Expressão Gênica , Humanos , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , MicroRNAs/metabolismoRESUMO
AIMS: Osteoarthritis (OA) is the leading cause of physical disability in middle-aged and elderly people globally. Previous studies have revealed that circular RNA (circRNA) is involved in the pathogenesis of OA. In this study, we studied the role of circ_0001846 in interleukin-1ß (IL-1ß)-induced OA progression. METHODS: Twenty-one patients with OA and 17 volunteers were recruited for the collection of articular cartilage tissues. The expression of circ_0001846, microRNA-149-5p (miR-149-5p) and Wingless-type MMTV integration site family, member 5B (WNT5B) was detected by quantitative real-time polymerase chain reaction. The protein expression was determined by western blot analysis. Cell viability, apoptosis, invasion and migration were demonstrated by cell counting kit-8, flow cytometry analysis, transwell invasion and wound-healing assays, respectively. The levels of IL-6 and tumor necrosis factor-α were detected by Enzyme-linked immunosorbent assay. The interaction between miR-149-5p and circ_0001846 or WNT5B was predicted by starbase online database, and proved by dual-luciferase reporter and RIP assays. RESULTS: Circ_0001846 and WNT5B expression were upregulated, while miR-149-5p expression was downregulated in articular cartilage tissues from patients with OA and IL-1ß-treated CHON-001 cells compared with normal articular cartilage tissues or untreated CHON-001 cells. Circ_0001846 expression was increased in IL-1ß-treated CHON-001 cell exosomes. Circ_0001846 knockdown reversed IL-1ß-mediated cell proliferation, apoptosis, migration, invasion, inflammation and extracellular matrix (ECM) degradation in CHON-001 cells. Additionally, circ_0001846 participated in IL-1ß-induced chondrocyte cell damage by sponging miR-149-5p. MiR-149-5p mediated IL-1ß-induced chondrocyte cell dysfunction by targeting WNT5B. Furthermore, circ_0001846 secretion was mediated by exosomes in IL-1ß-treated CHON-001 cells. CONCLUSION: Exosome-mediated transfer of circ_0001846 modulated IL-1ß-induced chondrocyte cell damage by miR-149-5p/WNT5B axis, providing a novel avenue for the therapy of OA.
Assuntos
Condrócitos/patologia , Interleucina-1beta/metabolismo , MicroRNAs/metabolismo , RNA Circular/metabolismo , Proteínas Wnt/metabolismo , Exossomos/genética , Exossomos/metabolismo , Regulação da Expressão Gênica/genética , Humanos , MicroRNAs/genética , Osteoartrite/genética , Osteoartrite/metabolismo , Osteoartrite/patologia , RNA Circular/genética , Proteínas Wnt/genéticaRESUMO
This research aimed at exploring the predictive value of 4-Hydroxyglutamate and miR-149-5p on eclampsia. Preeclampsia patients admitted to our hospital (n = 204), with 112 mild patients and 92 severe patients. Thereinto, pregnant women who underwent physical examination were regarded as a normal group (NG) (n = 100). Serum 4-Hydroxyglutamate levels and miR-149-5p in each group were detected. The serum 4-Hydroxyglutamate level in pregnant women in the NG was markedly lower than that in preeclampsia, while the miR-149-5p level was higher (p = 0.001). The serum 4-Hydroxyglutamate level in severe preeclampsia was higher than that in mild preeclampsia, while the miR-149-5p level was lower (p = 0.001). Partial thromboplastin time (APTT) and prothrombin time (PT) of preeclampsia patients were lower than those of the NG, while Fibrinogen (Fib) was higher (p = 0.001). With the aggravation of the condition of patients, PT, APTT decreased and Fib index increased. In preeclampsia patients, serum 4-Hydroxyglutamate was negatively correlated with PT and APTT, positively correlated with Fib content (p < 0.001); serum miR-149-5p was dramatically positively correlated with PT and APTT, negatively correlated with Fib content (p < 0.001). 4-Hydroxyglutamate and miR-149-5p were relevant to the occurrence time of preeclampsia; 4-Hydroxyglutamate, miR-149-5p and their combination could be used for preeclampsia diagnosis. According to the situation of newborn, they were divided into good and poor groups. The 4-Hydroxyglutamate level in the good group was lower than that in the poor group, while the miR-149-5p level was higher. The adverse prognosis of preeclampsia patients was predicted by 4-Hydroxyglutamate and miR-149-5p. 4-Hydroxyglutamate is highly expressed in preeclampsia, while miR-149-5p is low. Single and combined detection of 4-Hydroxyglutamate, miR-149-5p can be used for preeclampsia diagnosis and prediction.
Assuntos
Eclampsia/genética , Glutamatos/metabolismo , MicroRNAs/metabolismo , Adulto , Coagulação Sanguínea , Eclampsia/sangue , Eclampsia/diagnóstico , Feminino , Glutamatos/sangue , Humanos , Recém-Nascido , MicroRNAs/sangue , MicroRNAs/genética , Valor Preditivo dos Testes , Gravidez , Prognóstico , Curva ROCRESUMO
BACKGROUND: Circular RNAs (circRNAs) are related to oral squamous cell carcinoma (OSCC) progression. circRNA bicaudal D cargo adaptor 2 (circBICD2) has been reported to be abnormally expressed in OSCC. However, the function and mechanism of this circRNA in OSCC progression remain largely unknown. METHODS: circBICD2, microRNA-149-5p (miR-149-5p), and insulin-like growth factor 2 mRNA-binding protein 1 (IGF2BP1) abundances were examined via quantitative reverse transcription polymerase chain reaction or Western blot. The function of circBICD2 was investigated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), colony formation, flow cytometry, wound healing, transwell, specific kits, Western blot, and xenograft analyses. Dual-luciferase reporter analysis and RNA immunoprecipitation were carried out to analyze the binding interaction. RESULTS: circBICD2 expression was enhanced in OSCC tissues and cells. circBICD2 silence suppressed OSCC cell proliferation, migration, invasion, and glutaminolysis and facilitated apoptosis. miR-149-5p was targeted via circBICD2 and decreased in OSCC tissues and cells. miR-149-5p knockdown attenuated silence of circBICD2 on the influence of OSCC cell proliferation, apoptosis, migration, invasion, and glutaminolysis. IGF2BP1 was targeted via miR-149-5p, and circBICD2 could regulate IGF2BP1 via miR-149-5p. IGF2BP1 interference constrained OSCC cell proliferation, migration, invasion, and glutaminolysis and promoted apoptosis. circBICD2 silence reduced OSCC cell growth in xenograft model. CONCLUSION: circBICD2 knockdown repressed OSCC cell proliferation, migration, invasion, and glutaminolysis and increased apoptosis via modulating miR-149-5p/IGF2BP1 axis, which might act as a potential target for OSCC treatment.
Assuntos
Carcinoma de Células Escamosas , Neoplasias de Cabeça e Pescoço , MicroRNAs , Neoplasias Bucais , RNA Circular , Proteínas de Ligação a RNA , Carcinoma de Células Escamosas/genética , Movimento Celular/genética , Proliferação de Células/genética , Humanos , MicroRNAs/genética , Proteínas Associadas aos Microtúbulos , Neoplasias Bucais/genética , RNA Circular/genética , Proteínas de Ligação a RNA/genética , Carcinoma de Células Escamosas de Cabeça e PescoçoRESUMO
Parkinson's disease (PD) is a complex neurodegenerative disease in which the understanding of the underlying molecular mechanisms can be constructive in the diagnosis and treatment. Matrix metalloproteinase (MMPs) elevation and damage to the blood-brain barrier (BBB) are critical mechanisms involved in the PD separation. Studies have revealed that changes in miR-149-5p and CoQ10 are associated with BBB damage, and CoQ10 can affect the levels of some miRs. Hence, in the present study, we aimed to evaluate CoQ10 and miR-149-5p mimic on miR-149-5p, MMPs and TH expression, and behavioral functions of the PD models. PD was induced by injection of 6-OHDA into the rats' Medial Forbrain Bundle (MFB). The behavioral tests, including the Rotation test, Rotarod test, and Open field test, have been directed two weeks after PD induction. Next, the MiR-149-5p mimic (miR-mimic) and CoQ10 have been administered to rats. The same behavioral tests have been evaluated two weeks after administration to investigate the effect of miR-149-5p mimic and CoQ10. The rats were followed extra four weeks, and the behavioral tests have performed again. Finally, the expression of MMPs and miR-149-5p genes was measured using RT-qPCR, and tyrosine hydroxylase (TH) was assessed through immunohistochemistry analysis. According to the obtained results, the level of miR-149-5p has decreased, followed by PD induction in rats. RT-qPCR analysis has represented upregulation and downregulation of miR-149-5p and MMP-2,9, respectively, after miR-mimic and CoQ10 treatment. The treated rats have also represented improved motor function and increased TH + cells in the striatum according to the behavioral tests and immunohistochemistry assay. Taking together miR-149 and CoQ10 has shown to have an impressive potential to prevent damage to dopaminergic neurons caused by 6-OHDA injection through reducing MMP-2,9, increased TH expression, and improved motor function.
Assuntos
MicroRNAs , Doenças Neurodegenerativas , Fármacos Neuroprotetores , Doença de Parkinson , Animais , Modelos Animais de Doenças , Metaloproteinases da Matriz/genética , MicroRNAs/genética , MicroRNAs/metabolismo , Fármacos Neuroprotetores/farmacologia , Fármacos Neuroprotetores/uso terapêutico , Oxidopamina , Doença de Parkinson/tratamento farmacológico , Ratos , Ubiquinona/análogos & derivadosRESUMO
miR-149 is an miRNA with essential roles in carcinogenesis. This miRNA is encoded by the MIR149 gene on 2q37.3. The miR-149 hairpin produces miR-149-5p and miR-149-3p, which are the "guide" and the sister "passenger" strands, respectively. Deep sequencing experiments have shown higher prevalence of miR-149-5p compared with miR-149-3p. Notably, both oncogenic and tumor suppressive roles have been reported for miR-149-5p. In this review, we summarize the impact of miR-149-5p in the tumorigenesis and elaborate mechanisms of its involvement in this process in a variety of neoplastic conditions based on three lines of evidence, i.e., in vitro, in vivo and clinical settings.
Assuntos
Carcinogênese/genética , MicroRNAs/metabolismo , Animais , Carcinogênese/patologia , Linhagem Celular Tumoral , Modelos Animais de Doenças , Ontologia Genética , Redes Reguladoras de Genes , Genes Supressores de Tumor , Humanos , MicroRNAs/genéticaRESUMO
Circular RNAs (circRNAs) are involved in many courses of atherosclerosis and coronary artery disease (CHD). However, the role and effect of circRNAs in vascular restenosis after PCI remains unclear. Human aortic vascular smooth muscle cell (HA-VSMC) was cultured and stimulated with PDGF-BB. The expression profile of circRNAs in HA-VSMCs was screened using microarray analysis. A total 257 aberrantly expressed circRNAs were screened with 2 fold change. Has_circ_0113656 (also called circDHCR24) was validated by qRT-PCR to be significantly up-regulated in PDGF-BB induced HA-VSMCs. CircDHCR24 silencing obviously inhibited the proliferation, migration and phenotypic switch. Moreover, bioinformatics analysis predicted that miR-149-5p had complementary binding sites in 3'-UTR of circDHCR24. Luciferase reporter assay and RIP assay further verified the circDHCR24 acts as a spong for miR-149-5p in HA-VSMCs. Besides, bioinformatics analysis, luciferase reporter assay and RIP assay proved MMP9 was a directly target of miR-149-5p. Finally, cells were transfected with si-circDHCR24 with or without miR-149 inhibitor, and the results showed that co-transfection si-circDHCR24 and miR-149 inhibitor reversed the effect of si-circDHCR24 on cell proliferation, migration and phenotypic switch. Taken together, our study suggested for the first time that the knockdown of circDHCR24 alleviates HA-VSMCs proliferation, migration and phenotypic switching, thereby preventing vascular restenosis.
Assuntos
Aorta/citologia , Metaloproteinase 9 da Matriz/genética , MicroRNAs/genética , Músculo Liso Vascular/citologia , RNA Circular/genética , Aorta/metabolismo , Movimento Celular , Proliferação de Células , Técnicas de Silenciamento de Genes , Células HEK293 , Humanos , Músculo Liso Vascular/metabolismo , Transcriptoma , Regulação para CimaRESUMO
Circular RNAs (circRNAs) are novel noncoding RNAs (ncRNAs) with covalently closed-loop structures that play an essential regulatory role in diverse malignancies, including esophageal squamous cell cancer (ESCC). Circ_0000654 expression in ESCC and its mechanism of action remains unclear. Real-time PCR (RT-PCR) was employed to detect circ_0000654 and miR-149-5p expression in ESCC tissues. Circ_0000654 and miR-149-5p expression in ESCC cells was selectively regulated. Furthermore, interleukin 6 (IL-6) and signal transducer and activator of transcription 3 (STAT3) expression in cells was detected by RT-PCR and western blot analysis, while CCK8, BrdU, flow cytometry, and transwell assays were used to monitor cell proliferation, apoptosis, migration and invasion, respectively. The dual-luciferase reporter assay and RIP assay were used to verify the targeting relationship between circ_0000654 and miR-149-5p, miR-149-5p and IL-6. The function of circ_0000654 on ESCC cell proliferation and metastasis in vivo was examined using a subcutaneous xenograft model and a tail intravenous injection model in nude mice. Circ_0000654 was significantly upregulated in ESCC tissues and cell lines, and its high expression was remarkably associated with an increased T stage and local lymph node metastasis in ESCC patients. Circ_0000654 overexpression and knockdown experiments revealed that circ_0000654 regulated ESCC cell proliferation, migration, invasion, and apoptosis in vitro. Circ_0000654 was identified as a sponge of miR-149-5p and facilitated ESCC progression by indirectly activating the IL-6/STAT3 signaling pathway. Additionally, knocking down circ_0000654 strikingly repressed ESCC growth and metastasis in vivo. In summary, circ_0000654 functions as an oncogenic circRNA in ESCC and accelerates ESCC progression via adsorbing miR-149-5p and activating the IL-6/STAT3 signaling pathway.
Assuntos
Neoplasias Esofágicas/patologia , Carcinoma de Células Escamosas do Esôfago/patologia , MicroRNAs/genética , RNA Circular/genética , Idoso , Animais , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Neoplasias Esofágicas/genética , Neoplasias Esofágicas/metabolismo , Carcinoma de Células Escamosas do Esôfago/genética , Carcinoma de Células Escamosas do Esôfago/metabolismo , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Interleucina-6/genética , Interleucina-6/metabolismo , Masculino , Camundongos Endogâmicos BALB C , MicroRNAs/metabolismo , Pessoa de Meia-Idade , Fator de Transcrição STAT3/genética , Fator de Transcrição STAT3/metabolismo , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
BACKGROUND: Long non-coding RNAs (lncRNAs) are a class of endogenous non-coding RNAs of longer than 200 bp that play crucial roles in cancer biology. Here, we assessed the tumorigenic properties of a long noncoding RNA, MIAT, in non-small cell lung cancer (NSCLC). METHODS: Survival and clinicopathological analyses were done in a cohort of 80 patients with NSCLC. MIAT expression level were determined by real-time quantitative reverse transcriptase PCR (qRT-PCR). Dual luciferase reporter assays were employed to test the interaction between MIAT and miR-149-5p. Ectopic overexpression and shRNA-mediated knockdown of MIAT, CCK-8 and colony formation assays, Transwell migration and invasion in vitro, and in vivo tumorigenesis experiment were used to evaluate the function of MIAT. RESULTS: MIAT was significantly up-regulated in NSCLC tissues and cell lines, and was closely associated with advanced pathological stage and poor overall survival. Gain- and loss-of-function experiments in cell lines and mouse xenograft models showed that MIAT promoted the proliferation, migration, and invasion of NSCLC cells in vitro and accelerated tumor growth in vivo. Luciferase assay, western blotting, qRT-PCR, and rescue experiments showed that, mechanistically, MIAT could directly bind to miR-149-5p, and subsequently served as a sponge to increase the expression level of Forkhead box M1 (FOXM1). CONCLUSIONS: Our study reveals that MIAT acts as an oncogene in NSCLC via a novel MIAT/miR-149/FOXM1 axis, thus providing potential biomarkers and therapeutic targets for the management of NSCLC.