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1.
Beijing Da Xue Xue Bao Yi Xue Ban ; 55(2): 217-227, 2023 Apr 18.
Artigo em Chinês | MEDLINE | ID: mdl-37042131

RESUMO

OBJECTIVE: To identify and characterize read-through RNAs and read-through circular RNAs (rt-circ-HS) derived from transcriptional read-through hypoxia inducible factor 1α (HIF1α) and small nuclear RNA activating complex polypeptide 1 (SNAPC1) the two adjacent genes located on chromosome 14q23, in renal carcinoma cells and renal carcinoma tissues, and to study the effects of rt-circ-HS on biological behavior of renal carcinoma cells and on regulation of HIF1α. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) and Sanger sequencing were used to examine expression of read-through RNAs HIF1α-SNAPC1 and rt-circ-HS in different tumor cells. Tissue microarrays of 437 different types of renal cell carcinoma (RCC) were constructed, and chromogenic in situ hybridization (ISH) was used to investigate expression of rt-circ-HS in different RCC types. Small interference RNA (siRNA) and artificial overexpression plasmids were designed to examine the effects of rt-circ-HS on 786-O and A498 renal carcinoma cell proliferation, migration and invasiveness by cell counting kit 8 (CCK8), EdU incorporation and Transwell cell migration and invasion assays. RT-PCR and Western blot were used to exa-mine expression of HIF1α and SNAPC1 RNA and proteins after interference of rt-circ-HS with siRNA, respectively. The binding of rt-circ-HS with microRNA 539 (miR-539), and miR-539 with HIF1α 3' untranslated region (3' UTR), and the effects of these interactions were investigated by dual luciferase reporter gene assays. RESULTS: We discovered a novel 1 144 nt rt-circ-HS, which was derived from read-through RNA HIF1α-SNAPC1 and consisted of HIF1α exon 2-6 and SNAPC1 exon 2-4. Expression of rt-circ-HS was significantly upregulated in 786-O renal carcinoma cells. ISH showed that the overall positive expression rate of rt-circ-HS in RCC tissue samples was 67.5% (295/437), and the expression was different in different types of RCCs. Mechanistically, rt-circ-HS promoted renal carcinoma cell proliferation, migration and invasiveness by functioning as a competitive endogenous inhibitor of miR-539, which we found to be a potent post-transcriptional suppressor of HIF1α, thus promoting expression of HIF1α. CONCLUSION: The novel rt-circ-HS is highly expressed in different types of RCCs and acts as a competitive endogenous inhibitor of miR-539 to promote expression of its parental gene HIF1α and thus the proliferation, migration and invasion of renal cancer cells.


Assuntos
Carcinoma de Células Renais , Subunidade alfa do Fator 1 Induzível por Hipóxia , Neoplasias Renais , MicroRNAs , RNA Circular , Humanos , Carcinoma de Células Renais/patologia , Proliferação de Células , Hipóxia , MicroRNAs/genética , Invasividade Neoplásica/genética , RNA Circular/genética , RNA Circular/metabolismo , RNA Interferente Pequeno , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética
2.
Mol Biol Rep ; 48(11): 7059-7065, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34596809

RESUMO

BACKGROUND: Nostoc commune Vauch. is a nitrogen-fixing blue-green algae that expresses a large number of active molecules with medicinal properties. Our previous study found that a water stress protein (WSP1) from N. commune and its recombinant counterpart (Re-WSP1) exhibited significant anti-colon cancer activity both in vitro and in vivo. This study is to investigate the effects of Re-WSP1 on proliferation of colon cancer cells and to elucidate the relevant mechanisms. METHODS: Real-time quantitative PCR was used to detect the expression of miR-539 in colon cancer HT-29 and DLD1 cells. Colon cancer cells were transfected with miR-539 mimics and negative controls, and cell proliferation were detected by CCK8 and clonogenic assays. The target gene of miR-539 was predicted, and the dual luciferase reporter gene experiment was used to verify the target gene. After colon cancer cells were transfected with miR-539 mimics or inhibitors, the expression of target gene ß-catenin was detected by Western blot. miR-539 inhibitor confirmed cell proliferation. RESULTS: Re-WSP1 inhibited colon cancer cell growth in a dose-dependent manner. Re-WSP1 inhibited the expression of ß-catenin, which was partly reversed by LiCl treatment. Quantitative PCR analysis showed that the expression of miR-539 was significantly upregulated after Re-WSP1 treatment. Moreover, miR-539 negatively regulated the expression of ß-catenin by directly binding to the 3'UTR of ß-catenin mRNA. The cell growth inhibition and the decrease in ß-catenin expression induced by Re-WSP1 were significantly reversed by miR-539 inhibitor. CONCLUSION: Re-WSP1 suppresses colon cancer cell growth via the miR-539/ß-catenin axis.


Assuntos
Proteínas de Bactérias/farmacologia , Neoplasias do Colo/metabolismo , MicroRNAs/metabolismo , Proteínas de Neoplasias/metabolismo , Nostoc commune/genética , RNA Neoplásico/metabolismo , Transdução de Sinais/efeitos dos fármacos , beta Catenina/metabolismo , Proteínas de Bactérias/genética , Neoplasias do Colo/tratamento farmacológico , Neoplasias do Colo/genética , Neoplasias do Colo/patologia , Células HT29 , Humanos , MicroRNAs/genética , Proteínas de Neoplasias/genética , RNA Neoplásico/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/farmacologia , beta Catenina/genética
3.
Am J Physiol Cell Physiol ; 319(2): C381-C391, 2020 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-32491927

RESUMO

Several microRNAs (miRNAs or miRs) regulate cerebral ischemic injury outcomes; however, little is known about the role of miR-539-5p during cerebral ischemic injury or the postischemic state. Cerebral ischemic injury was modeled in vitro by exposing human cortical neurons to oxygen-glucose deprivation (OGD) and in vivo by occluding the middle cerebral artery (MCAO) in a rat model. The effects of miR-539-5p, histone deacetylase 1 (HDAC1), and early growth response 2 (EGR2) on cerebral ischemia were investigated using gain- and loss-of-function experiments. We identified changes in miR-539-5p, HDAC1, EGR2, and phosphorylated c-Jun NH2-terminal kinase (JNK). The interaction among miR-539-5p, HDAC1, and EGR2 was determined by dual luciferase reporter gene assay, chromatin immunoprecipitation, and coimmunoprecipitation. We also investigated the effects on cell viability and apoptosis and changes in inflammatory cytokine expression and spatial memory on MCAO rats. miR-539-5p and EGR2 were poorly expressed, while HDAC1 was highly expressed in OGD-treated HCN-2 cells. miR-539-5p targeted HDAC1, while HDAC1 prevented acetylation of EGR2 resulting in its downregulation and subsequent activation of the JNK pathway. Overexpression of miR-539-5p or EGR2 or silencing HDAC1 improved viability and reduced apoptosis of OGD-treated HCN-2 cells in vitro. Furthermore, overexpression of miR-539-5p improved spatial memory, while decreasing cell apoptosis and inflammation in MCAO rats. Collectively, these data suggest that miR-539-5p targets HDAC1 to upregulate EGR2, thus blocking the JNK signaling pathway, by which cerebral ischemic injury is alleviated.


Assuntos
Isquemia Encefálica/genética , Histona Desacetilase 1/genética , MicroRNAs/genética , Animais , Apoptose/genética , Isquemia Encefálica/patologia , Citocinas/metabolismo , Progressão da Doença , Proteína 2 de Resposta de Crescimento Precoce/genética , Regulação da Expressão Gênica/genética , Glucose/metabolismo , Humanos , Inflamação/genética , Artéria Cerebral Média/lesões , Artéria Cerebral Média/patologia , Neurônios/metabolismo , Neurônios/patologia , Ratos
4.
J Cell Mol Med ; 23(9): 5934-5948, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31298493

RESUMO

Prostate cancer (PCa) is the second leading cause of cancer-related death in males, primarily due to its metastatic potential. The present study aims to identify the expression of microRNA-539 (miR-539) in PCa and further investigate its functional relevance in PCa progression both in vitro and in vivo. Initially, microarray analysis was conducted to obtain the differentially expressed gene candidates and the regulatory miRNAs, after which the possible interaction between the two was determined. Next, ectopic expression and knock-down of the levels of miR-539 were performed in PCa cells to identify the functional role of miR-539 in PCa pathogenesis, followed by the measurement of E-cadherin, vimentin, Smad4, c-Myc, Snail1 and SLUG expression, as well as proliferation, migration and invasion of PCa cells. Finally, tumour growth was evaluated in nude mice through in vivo experiments. The results found that miR-539 was down-regulated and DLX1 was up-regulated in PCa tissues and cells. miR-539 was also found to target and negatively regulate DLX1 expression, which resulted in the inhibition of the TGF-ß/Smad4 signalling pathway. Moreover, the up-regulation of miR-539 or DLX1 gene silencing led to the inhibition of PCa cell proliferation, migration, invasion, EMT and tumour growth, accompanied by increased E-cadherin expression and decreased expression of vimentin, Smad4, c-Myc, Snail1 and SLUG. In conclusion, the overexpression of miR-539-mediated DLX1 inhibition could potentially impede EMT, proliferation, migration and invasion of PCa cells through the blockade of the TGF-ß/Smad4 signalling pathway, highlighting a potential miR-539/DLX1/TGF-ß/Smad4 regulatory axis in the treatment of PCa.


Assuntos
Proteínas de Homeodomínio/antagonistas & inibidores , MicroRNAs/genética , Neoplasias da Próstata/genética , Neoplasias da Próstata/patologia , Proteína Smad4/metabolismo , Fatores de Transcrição/antagonistas & inibidores , Fator de Crescimento Transformador beta/metabolismo , Animais , Caderinas/metabolismo , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Regulação para Baixo , Transição Epitelial-Mesenquimal/genética , Regulação Neoplásica da Expressão Gênica , Humanos , Metástase Linfática/genética , Masculino , Camundongos , Camundongos Nus , Pessoa de Meia-Idade , Invasividade Neoplásica/genética , Transplante de Neoplasias , Células PC-3 , Proteínas Proto-Oncogênicas c-myc/metabolismo , Fatores de Transcrição da Família Snail/metabolismo , Transplante Heterólogo , Vimentina/metabolismo
5.
J Cell Biochem ; 120(6): 10830-10846, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30706537

RESUMO

Papillary thyroid carcinoma (PTC) is the most common type of thyroid malignancy, with growing incidence every year. microRNAs (miRs) are known to regulate the physiological and pathological processes of cancers, such as proliferation, migration, invasion, survival, and epithelial-mesenchymal transition (EMT). Herein, this study aimed to investigate the effect of miR-539 on cell proliferation, apoptosis, and EMT by targeting secretory leukocyte protease inhibitor (SLPI) via the transforming growth factor ß1 (TGF-ß1)/Smads signaling pathway in PTC. First, PTC-related differentially expressed genes and regulatory miR were screened using bioinformatics analysis, dual luciferase reporter gene assay, and ribonucleoprotein immunoprecipitation, which identified the SLPI gene and the regulatory miR-539 for this study. We identified SLPI as a highly expressed gene in PTC tissues, and SLPI was targeted and negatively regulated by miR-539. Then, we introduced a series of miR-539 mimics, miR-539 inhibitors, and small interfering RNA against SLPI plasmids into CGTHW-3 cells to examine the effects of miR-539 and SLPI on the expression of TGF-ß1/Smads signaling pathway-, EMT-, and apoptosis-related factors, as well as cell proliferation, migration, invasion, and apoptosis. The obtained results indicated that CGTHW-3 cells treated with silenced SLPI or overexpressed miR-539 suppressed the cell proliferation, migration, invasion abilities, and resistance to apoptosis of PTC cells, corresponding to increased expression of Bcl-2-associated X protein, TGF-ß1, Sekelsky mothers against dpp 4, and epithelial cadherin, and decreased B cell lymphoma 2, Vimentin, and N-cadherin. Altogether, we concluded that overexpressed miR-539 could inhibit the PTC cell proliferation and promote apoptosis and EMT by targeting SPLI via activation of the TGF-ß1/Smads signaling pathway.


Assuntos
Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , Inibidor Secretado de Peptidases Leucocitárias/genética , Proteínas Smad/genética , Câncer Papilífero da Tireoide/genética , Neoplasias da Glândula Tireoide/genética , Fator de Crescimento Transformador beta1/genética , Adulto , Idoso , Antígenos CD/genética , Antígenos CD/metabolismo , Caderinas/genética , Caderinas/metabolismo , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Transição Epitelial-Mesenquimal/genética , Feminino , Humanos , Masculino , MicroRNAs/antagonistas & inibidores , MicroRNAs/metabolismo , Pessoa de Meia-Idade , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Inibidor Secretado de Peptidases Leucocitárias/metabolismo , Transdução de Sinais , Proteínas Smad/metabolismo , Câncer Papilífero da Tireoide/metabolismo , Câncer Papilífero da Tireoide/patologia , Neoplasias da Glândula Tireoide/metabolismo , Neoplasias da Glândula Tireoide/patologia , Fator de Crescimento Transformador beta1/metabolismo , Vimentina/genética , Vimentina/metabolismo , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismo
6.
J Cell Biochem ; 119(10): 8346-8358, 2018 11.
Artigo em Inglês | MEDLINE | ID: mdl-29893431

RESUMO

This study aims to explore the effects of miR-539 on osteoblast proliferation and differentiation and osteoclast apoptosis in a rat model of osteoporosis, and its mechanism involving the regulation of the AXIN1-mediated wingless-Int (Wnt) signaling pathway. A rat model of osteoporosis was successfully established by ovariectomy. With osteoblasts and osteoclasts of rats not receiving ovariectomy in the sham group as control, those of osteoporotic rats were treated with miR-539 inhibitor, miR-539 mimic, and AXIN1 shRNA. The expression of miR-53, AXIN1, the Wnt pathway related-genes, apoptosis related-genes, and osteogenic markers were measured by RT-qPCR and Western blot analysis, respectively. Alkaline phosphatase (ALP) activity in osteoblast and tartrate-resistant acid phosphatase (TRAP) activity in osteoclasts were determined after cell transfection. Osteoblast and osteoclast viability was assayed by CCK-8 assay. Cell cycle and apoptosis of osteoblasts and osteoclasts were detected by flow cytometry. Lastly, alizarin red S staining was used to detect mineralized nodules of osteoblasts. Firstly, we determined that miR-539 was down-regulated in osteoblast and osteoclast of osteoporotic rats and AXIN1 was negatively regulated by miR-539. Additionally, overexpression of miR-539 increased the expressions of ß-catenin, LEF1, c-myc, cyclin D1, RUNX2, BGP, BMP-2 in osteoblast as well as ß-catenin, RhoA, caspase-3, and Bcl-2 in osteoclasts. Finally, overexpression of miR-539 elevated ALP activity, proliferation, and mineralized nodules in osteoblast and osteoclast apoptosis, with reduced TRAP activity in osteoclasts. Our results demonstrate that miR-539 promotes osteoblast proliferation and differentiation as well as osteoclast apoptosis through the AXIN1-dependent Wnt signaling pathway in osteoporotic rats.


Assuntos
Proteína Axina/genética , MicroRNAs/genética , Osteoblastos/metabolismo , Osteoclastos/metabolismo , Osteoporose/genética , Via de Sinalização Wnt , Fosfatase Alcalina/genética , Fosfatase Alcalina/metabolismo , Animais , Antagomirs/genética , Antagomirs/metabolismo , Apoptose/genética , Proteína Axina/antagonistas & inibidores , Proteína Axina/metabolismo , Sequência de Bases , Densidade Óssea , Ciclo Celular/genética , Diferenciação Celular , Proliferação de Células , Modelos Animais de Doenças , Feminino , Regulação da Expressão Gênica , Vida Livre de Germes , MicroRNAs/antagonistas & inibidores , MicroRNAs/metabolismo , Mimetismo Molecular , Oligorribonucleotídeos/genética , Oligorribonucleotídeos/metabolismo , Osteoblastos/citologia , Osteoclastos/citologia , Osteoporose/etiologia , Osteoporose/metabolismo , Osteoporose/patologia , Ovariectomia/efeitos adversos , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Ratos , Fosfatase Ácida Resistente a Tartarato/genética , Fosfatase Ácida Resistente a Tartarato/metabolismo
7.
Biochem Biophys Res Commun ; 504(4): 784-791, 2018 10 12.
Artigo em Inglês | MEDLINE | ID: mdl-30217449

RESUMO

Liver cancer has been considered as one of the major leading causes of cancer-related mortality. The incidence of liver cancer tends to increase in less developed regions. Increasing evidences have demonstrated that mircoRNAs (miRNAs) play crucial roles in the modulation of tumor growth and progression. Whereas, the functional role of miR-539 in hepatocellular carcinoma (HCC) is not well established. In our present study, we sought to explore biological role of miR-539 in HCC progression. qRT-PCR was utilized to evaluate the expression level of miR-539. Immunoblotting analysis, qRT-PCR and luciferase reporter assays were used for the identification of the potential target of miR-539. Proliferation, migration and invasion assays and flow cytometric were performed to assess the biological functional role of miR-539. The molecular signaling pathways related to the integration of miR-539 were also evaluated. MiR-539 was reduced in human HCC. Mitogen-activated protein kinase 1, also known as MAP2K1 was verified as the target of miR-539. Overexpression of miR-539 inhibited migration, invasion and cell proliferation, while apoptosis rate was increased. Knockdown or overexpression of MAP2K1 in HCC cell transfected with ag-miR-539 or in-miR-539 indicated that miR-539 suppresses the progression of HCC by directly targeting and regulating MAP2K1. Our results reveal that miR-539 might be a tumor suppressor in HCC, supporting a potential target for advanced therapeutic strategy for this disease.


Assuntos
Carcinoma Hepatocelular/genética , Neoplasias Hepáticas/genética , MAP Quinase Quinase 1/genética , MicroRNAs/metabolismo , Apoptose/genética , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patologia , Movimento Celular/genética , Proliferação de Células/genética , Regulação para Baixo , Regulação Neoplásica da Expressão Gênica , Técnicas de Silenciamento de Genes , Genes Supressores de Tumor , Células Hep G2 , Humanos , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patologia , MAP Quinase Quinase 1/metabolismo , Sistema de Sinalização das MAP Quinases/genética
8.
Biochem Biophys Res Commun ; 464(4): 1128-1133, 2015 Sep 04.
Artigo em Inglês | MEDLINE | ID: mdl-26206083

RESUMO

MicroRNAs (miRNAs) are important regulators of multiple cellular processes, and aberrant miRNA expression has been observed in thyroid cancer. However, the role of miRNAs in thyroid cancer metastasis remains largely unknown. In the current study, we found that miR-539 plays a suppressor role in thyroid cancer cell migration and invasion. Luciferase reporter assays confirmed that miR-539 binding to the 3'-UTR region of CARMA1 inhibited the expression of CARMA1 in thyroid cancer cells. Further studies demonstrated that CARMA1 can significantly promote the migration and invasion of thyroid cancer cells. Interestingly, overexpression or knockdown of CARMA1 effectively blocked the effect of miR-539 on the migration and invasion of thyroid cancer cells. Furthermore, we showed that miR-539 expression was frequently downregulated and CARMA1 expression was significantly upregulated in thyroid cancer cell lines and thyroid cancer tissues compared with controls. Taken together, our data indicate that miR-539 is a novel regulator of migration and invasion in human thyroid cancer cells by targeting CARMA1.


Assuntos
Proteínas Adaptadoras de Sinalização CARD/genética , Guanilato Ciclase/genética , MicroRNAs/genética , Neoplasias da Glândula Tireoide/genética , Neoplasias da Glândula Tireoide/patologia , Linhagem Celular Tumoral , Movimento Celular/genética , Humanos , Invasividade Neoplásica , Neoplasias da Glândula Tireoide/metabolismo
9.
Mol Med Rep ; 30(1)2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38757308

RESUMO

Following the publication of this paper, it was drawn to the Editor's attention by a concerned reader that certain of the Transwell invasion assay data shown in Fig. 2C on p. 4921 were strikingly similar to data that had already been submitted for publication in different form in another article written by different authors at a different research institute. Owing to the fact that the contentious data in the above article had already been published prior to its submission to Molecular Medicine Reports, the Editor has decided that this paper should be retracted from the Journal. The authors were asked for an explanation to account for these concerns, but the Editorial Office did not receive a  reply. The Editor apologizes to the readership for any inconvenience caused. [Molecular Medicine Reports 17: 4917­4924, 2018; DOI: 10.3892/mmr.2018.8497].

10.
Cell Cycle ; 19(10): 1143-1157, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32308105

RESUMO

Differential expression of LINC00339 is involved in the malignancy of multiple human cancer types. Nonetheless, the expression profile, functions, and potential mechanisms of action of LINC00339 in gastric cancer are yet to be fully elucidated. This study aimed at measuring LINC00339 expression in gastric cancer and examining the prognostic significance of LINC00339 in patients with gastric cancer. The detailed functions of LINC00339 with regard to the aggressive characteristics of gastric cancer cells and the underlying molecular mechanisms were investigated. Here, we found that LINC00339 expression was aberrantly high in gastric cancer and significantly associated with lymph node metastasis, invasive depth, and TNM stage. Patients with gastric cancer in a LINC00339 high-expression group showed shorter overall survival than patients in a LINC00339 low-expression group. A knockdown of LINC00339 suppressed gastric cancer cell proliferation, migration, and invasion and induced apoptosis in vitro and slowed tumor growth in vivo. In terms of the mechanism, LINC00339 was found to act as a molecular sponge on microRNA-539 (miR-539). SRY-box 9 (SOX9) was confirmed as a direct target gene of miR-539 in gastric cancer cells. An miR-539 knockdown attenuated the effects of the LINC00339 knockdown on the malignant characteristics of gastric cancer cells. Overall, LINC00339 plays a critical role in the malignancy of gastric cancer by regulating SOX9 via sponging of miR­539. Our findings highlight the importance of the LINC00339-miR-539-SOX9 pathway in gastric cancer pathogenesis and may point to novel targets for the diagnosis, prognosis, and/or treatment of gastric cancer.


Assuntos
MicroRNAs/metabolismo , Oncogenes , RNA Longo não Codificante/metabolismo , Fatores de Transcrição SOX9/metabolismo , Transdução de Sinais/genética , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/patologia , Animais , Apoptose/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Feminino , Técnicas de Silenciamento de Genes , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , MicroRNAs/genética , Pessoa de Meia-Idade , Invasividade Neoplásica/genética , Prognóstico , RNA Longo não Codificante/genética , Fatores de Transcrição SOX9/genética , Transfecção , Carga Tumoral/genética , Regulação para Cima , Ensaios Antitumorais Modelo de Xenoenxerto
11.
Oncol Lett ; 16(2): 2693-2700, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30013665

RESUMO

Colorectal cancer (CRC) is the third most prevalent cancer and the fourth most common cause of cancer-associated mortality in males and females globally. Aberrant expression of microRNA-539 (miR-539) has been reported in multiple types of cancer. However, miR-539 expression, function and underlying mechanisms have not been clearly elucidated in CRC. In the present study, miR-539 expression was detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) in CRC tissues and cell lines. The effects of miR-539 on CRC cells were further examined in in vitro studies. In addition, the direct targets of miR-539 in CRC were investigated using bioinformatics, luciferase reporter assays, RT-qPCR and western blotting. miR-539 was revealed to be significantly downregulated in CRC cell lines and tissues. Decreased miR-539 expression was associated with lymph node metastasis and tumor-node-metastasis stage in patients with CRC. Functional assays revealed that the rescue of miR-539 expression attenuated CRC cell proliferation and invasion in vitro. Additionally, SRY-box 4 (SOX4) was validated as a direct target gene of miR-539 in CRC. Furthermore, SOX4 was revealed to be upregulated in CRC tissues at the mRNA and protein level. A significant negative correlation between miR-539 and SOX4 mRNA expression levels was observed in CRC tissues. Furthermore, upregulation of SOX4 partially restored the tumor suppressive effects of miR-539 on CRC cell proliferation and invasion. Taken together, this suggests that miR-539 may serve tumor-suppressive functions in CRC during the process of malignant transformation, by directly targeting SOX4. miR-539/SOX4-based targeted therapy may represent a potential novel treatment for patients with CRC.

12.
Exp Ther Med ; 15(3): 2681-2687, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29467860

RESUMO

The present study aimed to investigate the role and mechanism of microRNA-539 (miR-539) in rheumatoid arthritis (RA). A total of 68 RA patients and 46 osteoarthritis patients were enrolled into the current study. Peripheral blood and joint fluid were collected prior to treatment. Reverse transcription-quantitative polymerase chain reaction was performed to detect osteopontin (OPN) mRNA and miR-539 expression levels, while ELISA and western blot analysis were applied to detect OPN protein expression. In addition, bioinformatics analysis predicted that miR-539 directly targeted OPN, while dual-luciferase assay was used to validate this finding. Furthermore, agomiR-539 transfection and OPN knockdown by siRNA were conducted in MH7A cells, and MTT assay was used to detect MH7A cell proliferation. The results indicated that OPN was significantly increased in the blood and joint fluid of RA patients, while miR-539 expression was significantly decreased in the two types of specimens (P<0.05). Subsequent to silencing OPN by siRNA, the proliferation of MH7A cells was decreased (P<0.05). Following upregulation of miR-539, OPN expression was significantly decreased and cell proliferation was inhibited. Dual-luciferase assay revealed that miR-539 regulated OPN expression through complementary binding to 3'-untranslated region. OPN was also significantly increased in the blood and joint fluid of RA patients, which may be associated with the downregulation of miR-539. Thus, miR-539 may promote the development and progression of RA through regulating OPN.

13.
Mol Med Rep ; 17(4): 5611-5618, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29436648

RESUMO

Renal cell carcinoma (RCC) is one of the most common urinary malignancies with a high rate of morbidity. MicroRNAs (miRNAs) have been shown to be critical post­transcriptional regulators in tumorigenesis. The present study aimed to investigate the effect of miRNA (miR)­539 on the proliferation and apoptosis of RCC. The expression of miR­539 and high mobility group AT­hook 2(HMGA2) were examined in clinical RCC specimens. The 786­O RCC cell line was also used and was transfected with miR­539 mimics or inhibitors. The correlation between miR­539 and HMGA2 was confirmed using a luciferase reporter assay. Cell viability and apoptosis were detected using MTT and flow cytometry assays. The protein levels of HMGA2, AKT, phosphorylated (p)­AKT, mammalian target of rapamycin (mTOR) and p­mTOR were analyzed using western blot analysis. The results revealed that miR­539 was negatively correlated with the expression of HMGA2 in clinical RCC specimens. Further experiments identified HMGA2 as a direct target of miR­539. The overexpression of miR­539 downregulated the expression of HMGA2, reduced cell proliferation and promoted cell apoptosis, whereas the knockdown of miR­539 led to the opposite results. miR­539 also suppressed the phosphorylation of AKT and mTOR, without altering the levels of total AKT and mTOR. Taken together, the results of the present study indicated that miR­539 negatively regulated the expression of HMGA2 in clinical specimens and in vitro. miR539 inhibited cell proliferation and induced apoptosis in RCC cells. This regulatory effect of miR­539 may be associated with the AKT signaling pathway. Therefore, miR­539 may be used as a biomarker for predicting the progression of RCC.


Assuntos
Carcinoma de Células Renais/genética , Regulação Neoplásica da Expressão Gênica , Proteína HMGA2/genética , Neoplasias Renais/genética , MicroRNAs/genética , Interferência de RNA , Adulto , Idoso , Apoptose/genética , Carcinoma de Células Renais/metabolismo , Carcinoma de Células Renais/patologia , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Feminino , Genes Reporter , Humanos , Imuno-Histoquímica , Neoplasias Renais/metabolismo , Neoplasias Renais/patologia , Masculino , Pessoa de Meia-Idade , Gradação de Tumores , Metástase Neoplásica , Estadiamento de Neoplasias , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais
14.
Mol Med Rep ; 17(4): 4917-4924, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29393438

RESUMO

Bladder cancer is the most frequent malignancy of the urinary tract and the seventh most common cancer worldwide. The abnormal expression of microRNAs has been frequently observed in various types of human cancers, including bladder cancer. In addition, an increasing body of evidence has demonstrated that microRNAs are potential targets for cancer diagnosis, treatments and prognosis. The aim of the present study was to investigate the expression patterns and potential roles of microRNA­539 (miR­539) in bladder cancer and its underlying mechanism. Reverse transcription­quantitative polymerase chain reaction (RT­qPCR) was performed to detect miR­539 expression in the bladder cancer tissues and cell lines. Following transfection, MTT and cell invasion assays were used to investigate the effects of miR­539 overexpression or IGF1R underexpression on bladder cancer cell proliferation and invasion. Bioinformatics analysis, a luciferase reporter assay, RT­qPCR and western blot analysis were utilized to determine the potential targets of miR­539 in bladder cancer. The results revealed that miR­539 levels were relatively decreased in bladder cancer tissues and cell lines when compared with those observed in the matched adjacent normal bladder tissues and normal bladder epithelial cell line. miR­539 expression was associated with the tumor stage and lymph node metastasis of patients with bladder cancer. In addition, the expression of miR­539 suppressed bladder cancer cell proliferation and invasion. Insulin like growth factor 1 receptor (IGF­1R) was identified as a direct target of miR­539, and miR­539 was also observed to regulate the protein kinase B and extracellular signal­regulated kinases signaling pathways. IGF­1R was markedly upregulated in bladder cancer tissues and negatively associated with miR­539 expression levels. Furthermore, IGF­1R knockdown in bladder cancer cells significantly inhibited cell proliferation and invasion. To the best of our knowledge, these results demonstrated for the first time that miR­539 may act as a tumor suppressor and serve important roles in tumorigenesis and progression of bladder cancer. Thus, miR­539/IGF­1R may be a potential therapeutic target for the treatment of bladder cancer.


Assuntos
Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , Interferência de RNA , Receptores de Somatomedina/genética , Neoplasias da Bexiga Urinária/genética , Regiões 3' não Traduzidas , Adulto , Idoso , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Feminino , Genes Reporter , Humanos , Masculino , Pessoa de Meia-Idade , Gradação de Tumores , Metástase Neoplásica , Estadiamento de Neoplasias , Proteínas Proto-Oncogênicas c-akt/metabolismo , Receptor IGF Tipo 1 , Transdução de Sinais , Carga Tumoral , Neoplasias da Bexiga Urinária/metabolismo , Neoplasias da Bexiga Urinária/patologia
15.
Journal of Clinical Surgery ; (12): 1160-1163, 2023.
Artigo em Chinês | WPRIM | ID: wpr-1019279

RESUMO

Objective To investigate the expression and clinical significance of microRNA(miR)-539-5p,Caspase activity and apoptosis inhibitory factor 1(CAAP1)in gastric cancer tissue.Methods The gastric cancer tissue specimens and corresponding adjacent tissues of gastric cancer patients(102 cases)who underwent surgery in Xi'an Ninth Hospita from January 2018 to December 2019 were collected,and the expression levels of miR-539-5p and CAAP1 in the tissues were detected and compared.The relationship between the expression of miR-539-5p and CAAP1 and clinicopathological parameters and prognosis was explored.The correlation between miR-539-5p and CAAP1 expression was analyzed by Pearson method.Kaplan-Meier method was used to analyze the survival time of gastric cancer patients.Cox regression model was used to analyze the prognostic factors of patients with gastric cancer.Results The expression level of miR-539-5p in gastric cancer tissue lower than that in adjacent tissue,and the expression level of CAAP1 higher than that in adjacent tissue(P<0.05).There was a negative correlation between the expression levels of miR-539-5p and CAAP1 in gastric cancer tissue(P=0.000).Further analysis showed that the expression of miR-539-5p and CAAP1 in gastric cancer tissue was greatly correlated with TNM stage,depth of invasion,lymph node metastasis and tumor size(P<0.05).The 3-year survival rate of miR-539-5p high expression group was higher than that of low expression group(P=0.045).The 3-year survival rate of CAAP1 high expression group which was lower than low expression group(P=0.015).Cox regression showed that TNM stage,depth of invasion,lymph node metastasis and miR-539-5p and CAAP1 expression were prognostic factors in patients with gastric cancer(P<0.05).Conclusion The decreased expression level of miR-539-5p and increased expression level of CAAP1 in gastric cancer tissues were related to TNM staging,depth of invasion,lymph node metastasis and tumor size,and they are closely related to the prognosis of patients.

16.
Artigo em Chinês | WPRIM | ID: wpr-986842

RESUMO

OBJECTIVE@#To identify and characterize read-through RNAs and read-through circular RNAs (rt-circ-HS) derived from transcriptional read-through hypoxia inducible factor 1α (HIF1α) and small nuclear RNA activating complex polypeptide 1 (SNAPC1) the two adjacent genes located on chromosome 14q23, in renal carcinoma cells and renal carcinoma tissues, and to study the effects of rt-circ-HS on biological behavior of renal carcinoma cells and on regulation of HIF1α.@*METHODS@#Reverse transcription-polymerase chain reaction (RT-PCR) and Sanger sequencing were used to examine expression of read-through RNAs HIF1α-SNAPC1 and rt-circ-HS in different tumor cells. Tissue microarrays of 437 different types of renal cell carcinoma (RCC) were constructed, and chromogenic in situ hybridization (ISH) was used to investigate expression of rt-circ-HS in different RCC types. Small interference RNA (siRNA) and artificial overexpression plasmids were designed to examine the effects of rt-circ-HS on 786-O and A498 renal carcinoma cell proliferation, migration and invasiveness by cell counting kit 8 (CCK8), EdU incorporation and Transwell cell migration and invasion assays. RT-PCR and Western blot were used to exa-mine expression of HIF1α and SNAPC1 RNA and proteins after interference of rt-circ-HS with siRNA, respectively. The binding of rt-circ-HS with microRNA 539 (miR-539), and miR-539 with HIF1α 3' untranslated region (3' UTR), and the effects of these interactions were investigated by dual luciferase reporter gene assays.@*RESULTS@#We discovered a novel 1 144 nt rt-circ-HS, which was derived from read-through RNA HIF1α-SNAPC1 and consisted of HIF1α exon 2-6 and SNAPC1 exon 2-4. Expression of rt-circ-HS was significantly upregulated in 786-O renal carcinoma cells. ISH showed that the overall positive expression rate of rt-circ-HS in RCC tissue samples was 67.5% (295/437), and the expression was different in different types of RCCs. Mechanistically, rt-circ-HS promoted renal carcinoma cell proliferation, migration and invasiveness by functioning as a competitive endogenous inhibitor of miR-539, which we found to be a potent post-transcriptional suppressor of HIF1α, thus promoting expression of HIF1α.@*CONCLUSION@#The novel rt-circ-HS is highly expressed in different types of RCCs and acts as a competitive endogenous inhibitor of miR-539 to promote expression of its parental gene HIF1α and thus the proliferation, migration and invasion of renal cancer cells.


Assuntos
Humanos , Carcinoma de Células Renais/patologia , Proliferação de Células , Hipóxia , Neoplasias Renais , MicroRNAs/genética , Invasividade Neoplásica/genética , RNA Circular/metabolismo , RNA Interferente Pequeno , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética
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