RESUMEN
The objective of this study was to determine the regulatory mechanism of MAGI2-AS3 in clear cell renal cell carcinoma (ccRCC), thereby supplying a new insight for ccRCC treatment. Expression data in TCGA-KIRC were obtained. Target gene lncRNA for research was determined using expression analysis and clinical analysis. lncRNA's downstream regulatory miRNA and mRNA were predicted by bioinformatics databases. ccRCC cell malignant phenotypes were detected via CCK-8, colony formation, Transwell migration, and invasion assays. The targeting relationship between genes was assessed through dual-luciferase reporter gene analysis. Kaplan-Meier (K-M) analysis was carried out to verify the effect of MAGI2-AS3, miR-629-5p, and PRDM16 on the survival rate of ccRCC patients. MAGI2-AS3 expression in ccRCC tissue and cells was shown to be markedly decreased and its expression to continuously decline with tumor progression. MAGI2-AS3 suppresses ccRCC proliferation and migration. Dual-luciferase assay showed that MAGI2-AS3 binds miR-629-5p and that miR-629-5p binds PRDM16. In addition, functional experiments showed that MAGI2-AS3 facilitates PRDM16 expression by repressing miR-629-5p expression, thereby suppressing ccRCC cell aggression. K-M analysis showed that upregulation of either MAGI2-AS3 or PRDM16 significantly improves ccRCC patient survival, while upregulation of miR-629-5p has no significant impact. MAGI2-AS3 sponges miR-629-5p to modulate PRDM16 to mediate ccRCC development. Meanwhile, the MAGI2-AS3/miR-629-5p/PRDM16 axis, as a regulatory pathway of ccRCC progression, may be a possible therapeutic target and prognostic indicator of ccRCC.
Asunto(s)
Carcinoma de Células Renales , Neoplasias Renales , MicroARNs , ARN Largo no Codificante , Humanos , Proteínas Adaptadoras Transductoras de Señales , Carcinoma de Células Renales/genética , Proteínas de Unión al ADN , Guanilato-Quinasas/genética , Neoplasias Renales/genética , MicroARNs/genética , ARN Largo no Codificante/genética , Factores de TranscripciónRESUMEN
Proven by publications, long non-coding RNAs (lncRNAs) play critical roles in the development of clear cell renal cell carcinoma (ccRCC). Although lncRNA LINC00565 has been implicated in the progression of various cancers, its biological effects on ccRCC remain unknown. This study aimed to investigate the biological functions of LINC00565, as well as its potential mechanism in ccRCC. Here, the expression data of mature microRNAs (miRNAs) (normal: 71, tumor: 545), messenger RNAs (mRNAs), and lncRNAs (normal: 72, tumor: 539) of ccRCC were acquired from The Cancer Genome Atlas (TCGA) database and subjected to differential expression analysis. Quantitative reverse transcriptase polymerase chain reaction analyzed the expression levels of LINC00565, miR-532-3p, and ADAM19 mRNA. TCGA database, dual-luciferase report detection, and Argonaute 2 RNA immunoprecipitation were utilized to confirm the relationships between LINC00565 and miR-532-3p and between miR-532-3p and ADAM19, respectively. The progression of ccRCC cells was determined via CCK-8, colony formation, scratch healing, and transwell assays. Western blot was applied to detect the protein levels of epithelial-mesenchymal transition markers and ADAM19. We herein suggested that LINC00565 was prominently upregulated in ccRCC tissues and cells. Knockdown of LINC00565 repressed cell progression. We further predicted and validated miR-532-3p as a target of LINC00565, and miR-532-3p could target ADAM19. Knockdown of LINC00565 resulted in ADAM19 level downregulation in ccRCC cells and suppressed miR-532-3p could restore ADAM19 level. Thus, the three RNAs constructed a ceRNA network. Overexpressed ADAM19 could eliminate the anticancer effects caused by knocking down LINC00565 on ccRCC cells. In conclusion, LINC00565 upregulated ADAM19 via absorbing miR-532-3p, thereby facilitating the progression of ccRCC cells.
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Carcinoma de Células Renales , Neoplasias Renales , MicroARNs , ARN Largo no Codificante , Humanos , Carcinoma de Células Renales/genética , Carcinoma de Células Renales/patología , ARN Largo no Codificante/genética , ARN Largo no Codificante/metabolismo , Línea Celular Tumoral , Proliferación Celular , MicroARNs/genética , MicroARNs/metabolismo , Neoplasias Renales/genética , Neoplasias Renales/patología , Movimiento Celular/genética , Regulación Neoplásica de la Expresión Génica , Proteínas ADAM/genética , Proteínas ADAM/metabolismoRESUMEN
BACKGROUND: The molecular heterogeneity of clear cell renal cell carcinoma (ccRCC) leads to a high mortality of the disease, which seriously threatens the life of patients. Therefore, this study explored the functional significance and mechanism of microRNA-155-5p and nuclear receptor subfamily 3 group C member 2 (NR3C2) in the regulation of ccRCC. METHODS: Expression levels of microRNA-155-5p and NR3C2 mRNA in ccRCC cells were analyzed by qRT-PCR, and the protein expression of NR3C2 in human ccRCC cells was measured by Western blot. Biological functions were determined through a series of in vitro experiments. The interaction between microRNA-155-5p and NR3C2 was tested by luciferase reporter gene assay. In addition, the effect of overexpressed or silenced microRNA-155-5p on cell phenotypes was evaluated in ccRCC cells. RESULTS: Experimental data suggested that overexpression or silencing of microRNA-155-5p in ccRCC could boost or suppress cancer cell proliferation and other malignant behaviors. Rescue experiments revealed that microRNA-155-5p facilitated the proliferation, migration, and invasion and suppressed the apoptosis of ccRCC by directly inhibiting the expression of NR3C2. CONCLUSIONS: This is the first study to generate new insights into the role of microRNA-155-5p/NR3C2 interaction in promoting the process of ccRCC, and it is possible to bring a turning point for the treatment of ccRCC.
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Carcinoma de Células Renales , Neoplasias Renales , MicroARNs , Receptores de Mineralocorticoides , Carcinoma de Células Renales/genética , Carcinoma de Células Renales/metabolismo , Carcinoma de Células Renales/patología , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Femenino , Humanos , Neoplasias Renales/genética , Neoplasias Renales/metabolismo , Neoplasias Renales/patología , Masculino , MicroARNs/genética , MicroARNs/metabolismo , Receptores de Mineralocorticoides/genética , Receptores de Mineralocorticoides/metabolismoRESUMEN
Prostate cancer (PCa) is one of the most prevalent malignant tumours. The alternation of microRNAs (miRNAs) expression is associated with prostate cancer progression, whereas its way to influence progression of prostate cancer remains elusive. The expression levels of PRDM16 mRNA and miR-372-3p in PCa cell lines were analysed using qRT-PCR. The protein expression of PRDM16 in PCa cell lines was also analysed using Western blot. CCK-8, wound healing and Transwell assays were applied to examine cell proliferation, migration, and invasion in prostate cancer cells, respectively. Dual-luciferase reporter assay was utilised to validate the interaction between miR-372-3p and PRDM16. In the present study, markedly decreased PRDM16 mRNA and protein expression levels were observed in prostate cancer cells. PRDM16 overexpression hampered cellular proliferation, migration, and invasion, while silencing PRDM16 had the opposite effect. Moreover, miR-372-3p could target the regulation expression of PRDM16. Rescue experiments demonstrated that upregulating miR-372-3p conspicuously restored the inhibitory effect of increased PRDM16 on cell proliferation, migration, and invasion in PCa. Overall, our study clarifies the biological role of miR-372-3p/PRDM16 axis in prostate cancer progression, which may be effective biomarkers for clinical treatment of prostate cancer.
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Objectives: To evaluate the clinical efficacy of neoadjuvant chemohormonal therapy (NCHT) combined with laparoscopic radical prostatectomy in high-risk prostate cancer (PCa). Methods: A randomized controlled trial was used in this study. Eighty patients with high-risk PCa treated in Tangshan Gongren Hospital from January 2017 to January 2019 were selected and randomly divided into two groups. The control group was given neoadjuvant endocrine therapy, while the research group was added NCHT to the control group. Three months later, the patients of two groups underwent laparoscopic radical prostatectomy. The changes of surgical indicators, adverse drug reactions, incidence of lower urinary tract symptoms, biochemical recurrence rate after follow-up, PSA progression-free survival and incidence of surgical complications were compared between the two groups. Results: After NCHT, the PSA level and prostate volume in the research group decreased significantly than those in the control group (P = 0.00). Surgical duration, postoperative hospital stay and retention time of drainage tube were significantly shorter and intraoperative blood loss was significantly less in the research group than those in the control group (P = 0.00). The incidence of lower urinary tract symptoms, biochemical recurrence and surgical complications in the research group were significantly lower than those in the control group, and the early recovery rate of urinary control and progression-free survival were significantly better than those in the control group (P < 0.05). Conclusion: NCHT combined with laparoscopic radical prostatectomy is a safe and effective treatment for high-risk PCa, which is worthy of promotion in clinical practice.
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BACKGROUND: MicroRNAs are significantly involved in tumorigenesis and progression of glioma. However, the critical part they play in glioma have not been fully elaborated. miR-491 and Tripartite motif containing 28 (TRIM28) are reported to aberrantly express in glioblastoma multiforme (GBM). Here, we detected miR-491 and TRIM28 expression and function in glioma cells. METHODS: We analyzed miR-491 expressions in 20 primary human GBM tissues and 6 control brain tissues by qRT-PCR assays and searched for The Cancer Genome Atlas (TCGA) database. Then we predicted possible mRNA target of miR-491 by TargetScan/MicroRNA and confirmed it via luciferase reporter assays. Knock-down of miR-491 and transfection of pLenti-TRIM28 were performed in U251 and U87 cells. Proliferation ability was examined by MTT and clone formation assays. RESULTS: miR-491 expression was obviously reduced in GBM cells and tissues. There was a positive correlation between the down-regulation of miR-491 and poor prognosis. Spearman's correlation analysis demonstrated that miR-491 expression was negatively correlated with TRIM28 protein level. Possible mRNA binding sites of miR-491 predicted by TargetScan/MicroRNA were proved by luciferase assays. Clone formation and MTT assays indicated that up-regulation of miR-491 inhibited the proliferation of glioma cells. CONCLUSIONS: miR-491 regulates glioma cells proliferation in vitro by targeting TRIM28.
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Neoplasias Encefálicas/genética , Glioblastoma/genética , MicroARNs/genética , Proteínas Represoras/genética , Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/patología , Línea Celular Tumoral , Proliferación Celular , Regulación hacia Abajo , Regulación Neoplásica de la Expresión Génica , Glioblastoma/metabolismo , Glioblastoma/patología , Humanos , MicroARNs/metabolismo , Pronóstico , Proteínas Represoras/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Proteína 28 que Contiene Motivos TripartitoRESUMEN
BACKGROUND: The inflammatory response is closely related to cancer progression and prognosis. The aim of this study was to determine the prognostic value of preoperative inflammatory markers among different molecular subtypes of lower-grade glioma (LGG). METHODS: We performed a retrospective analysis of 214 patients with LGG from 2001 to 2013, evaluating the effect of the neutrophil/lymphocyte ratio (NLR), lymphocyte/monocyte ratio (LMR), platelet/lymphocyte ratio (PLR) and derived NLR (dNLR) on prognosis among different molecular subtypes. Isocitrate dehydrogenase (IDH) and telomerase reverse transcriptase (TERT) promotor mutations were detected by gene sequencing, and Chromosome arms 1p and 19q (1p/19q) codeletion was estimated via fluorescence in situ hybridization. RESULTS: Survival analysis showed that a high NLR, low LMR, and high dNLR were associated with poor prognosis, while the PLR had no prognostic significance. The subsequent molecular subtype analysis indicated that a high NLR and dNLR predicted worse survival in the IDH mutation only group, a high NLR and PLR predicted worse survival in the IDH and TERT promoter mutation group, and a high PLR was associated with shorter survival in the triple-positive group. Furthermore, univariate and multivariate Cox regression analysis suggested that the dNLR was an independent prognostic factor for LGG. Finally, the prognostic nomogram was developed by integrating the inflammatory marker dNLR and independent clinical risk factors. CONCLUSION: The results of this study indicated that a high dNLR was an independent risk factor for overall survival rates in patients with LGG, which may increase prognostic accuracy and improve patient outcomes.
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Glioma , Biomarcadores de Tumor/genética , Glioma/diagnóstico , Glioma/genética , Glioma/cirugía , Humanos , Hibridación Fluorescente in Situ , Isocitrato Deshidrogenasa/genética , Clasificación del Tumor , Pronóstico , Regiones Promotoras Genéticas , Estudios Retrospectivos , Telomerasa/genéticaRESUMEN
Renal cell carcinoma (RCC) is a highly recurrent aggressive tumor. This study works for the regulation of miR-21-5p on RCC cell functions and novel ideas for therapies of RCC. Isoform expression quantification data were offered by The Cancer Genome Atlas Kidney Renal Clear Cell Carcinoma (TCGA-KIRC) to investigate differentially expressed miRNAs. The way miR-21-5p works on biological functions of RCC was examined with MTT and Transwell assays. The downstream targets of miR-21-5p were predicted using bioinformatics analysis. The binding of two researched objects was verified by the dual-luciferase method. TCGA data manifested a considerably high level of miR-21-5p in RCC tissue, while ARHGAP24 was significantly lowly expressed. miR-21-5p bound ARHGAP24 and stimulated RCC cell functions, whereas ARHGAP24 mimic could reverse such promotion. This work observed miR-21-5p, a stimulator in RCC, and it deteriorated this cancer via repressing its downstream target gene ARHGAP24 expression.
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Carcinoma de Células Renales , Neoplasias Renales , MicroARNs , Carcinoma de Células Renales/genética , Carcinoma de Células Renales/metabolismo , Carcinoma de Células Renales/patología , Línea Celular Tumoral , Proliferación Celular/genética , Regulación hacia Abajo , Proteínas Activadoras de GTPasa/genética , Proteínas Activadoras de GTPasa/metabolismo , Regulación Neoplásica de la Expresión Génica , Humanos , Neoplasias Renales/genética , Neoplasias Renales/metabolismo , Neoplasias Renales/patología , MicroARNs/genéticaRESUMEN
Prostate cancer (PC) is the most common malignancy in men. As per recent findings, microRNA-300 (miR-300) were found to be overexpressed in numerous types of cancers. In this study, we aimed to explore the effects of miR-300 on the adhesion, invasion, and migration of PC cells by targeting Disabled 1 (DAB1). Firstly, the regulatory role of miRNAs on DAB1 was predicted by screening PC-related differentially expressed genes (DEGs). Immunohistochemistry was applied to determine the positive protein expression of DAB1, after which the target relationship between miR-300 and DAB1 was examined. Loss-of-function and gain-of-function experiments were conducted to determine cell proliferation, adhesion, migration, invasion capability, and cell cycle of PC cells. Our data illustrated that DAB1 had a low expression, while miR-300 was expressed at a relatively high level in PC tissues. Moreover, our clinicopathological analysis revealed that there was a correlation between miR-300 and tumor, node, metastases stage, Gleason score, and lymph node metastasis of PC patients. DAB1 was also found to be poorly expressed in PC based on the findings from the microarray analysis. The results from dual-luciferase reporter gene assay corroborated that miR-300 interacts with DAB1. Importantly, overexpression of miR-300 and/or si-DAB1 resulted in the enhancement of RAC1, MMP2, MMP9, CyclinD1, and CyclinE expressions, whereas the expression of DAB1 and Rap was reduced in PC cells, thus suggesting that down-regulated miR-300 suppressed proliferation, adhesion, migration, and invasion of PC cells. Collectively, our results provided evidence that down-regulation of miR-300 inhibits the adhesion, migration, and invasion of PC cells.
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Proteínas Adaptadoras Transductoras de Señales/genética , Adhesión Celular/genética , Movimiento Celular/genética , MicroARNs/genética , Proteínas del Tejido Nervioso/genética , Neoplasias de la Próstata/genética , Adulto , Anciano , Anciano de 80 o más Años , Línea Celular Tumoral , Proliferación Celular/genética , Regulación hacia Abajo/genética , Regulación Neoplásica de la Expresión Génica/genética , Humanos , Metástasis Linfática/genética , Metástasis Linfática/patología , Masculino , Persona de Mediana Edad , Células PC-3 , Próstata/patología , Neoplasias de la Próstata/patologíaRESUMEN
Iron overload has been regarded as a common cause for refractory epilepsies in patients after hemorrhagic strokes. This study is to examine the potential epilepsy control effect of deferoxamine (DFO), an iron chelator, on a ferric chloride-induced epilepsy rat model. Twenty four rats were divided into 4 groups: group I is blank control group, group II is sham group with intracortical injection of saline, group III is epilepsy group with intracortical injection of iron and saline treatment, group IV is treatment group with intracortical injection of iron and DFO treatment. For the DFO intervention group, a daily dose of 100mg/kg DFO via peritoneal injection was applied for 14days. Outcomes were evaluated by behavioral study, electroencephalography (EEG), magnetic resonance imaging (MRI) scan and tissue analysis. Epilepsies according to behavioral observations and EEG analysis were significantly suppressed after intervention of DFO. Reduction of iron content in the brain cortex was proved by diminished low signal area on T2-MRI images (p=0.006) and tissue analysis (p<0.001), simultaneously the superoxide dismutase (SOD) activity increased (p<0.001). Western blot analysis demonstrated the decreasing of local transferrin after DFO treatment. DFO is efficient at Fe clearance, thus helpful in epilepsy control. This finding implies potential therapeutic value of DFO in patients with refractory epilepsy after hemorrhagic stroke.
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Anticonvulsivantes/farmacología , Deferoxamina/farmacología , Epilepsia/tratamiento farmacológico , Sideróforos/farmacología , Animales , Western Blotting , Encéfalo/diagnóstico por imagen , Encéfalo/efectos de los fármacos , Encéfalo/patología , Encéfalo/fisiopatología , Cloruros , Modelos Animales de Enfermedad , Evaluación Preclínica de Medicamentos , Electroencefalografía , Epilepsia/diagnóstico por imagen , Epilepsia/patología , Epilepsia/fisiopatología , Compuestos Férricos , Ferritinas/metabolismo , Hierro/metabolismo , Imagen por Resonancia Magnética , Masculino , Distribución Aleatoria , Ratas Sprague-Dawley , Superóxido Dismutasa/metabolismo , Transferrina/metabolismoRESUMEN
Tissue engineering is the application of interdisciplinary knowledge in the building and repairing of tissues. Generally, an engineered tissue is a combination of living cells and a support structure called a scaffold. The scaffold provides support for bone-producing cells and can be used to heal or replace a defective bone. In this chapter, a novel bone scaffold design approach based on shape function and an all-hexahedral mesh refinement method is presented. Based on the shape function in the finite element method, an all-hexahedral mesh is used to design a porous bone scaffold. First, the individual pore based on the subdivided individual element is modeled; then, the Boolean operation union among the pores is used to generate the whole pore model of TE bone scaffold; finally, the bone scaffold which contains various irregular pores can be modeled by the Boolean operation difference between the solid model and the whole pore model. From the SEM images, the pore size distribution in the native bone is not randomly distributed and there are gradients for pore size distribution. Therefore, a control approach for pore size distribution in the bone scaffold based on the hexahedral mesh refinement is also proposed in this chapter. A well-defined pore size distribution can be achieved based on the fact that a hexahedral element size distribution can be obtained through an all-hexahedral mesh refinement and the pore morphology and size are under the control of the hexahedral element. The designed bone scaffold can be converted to a universal 3D file format (such as STL or STEP) which could be used for rapid prototyping (RP). Finally, 3D printing (Spectrum Z510), a type of RP system, is adopted to fabricate these bone scaffolds. The successfully fabricated scaffolds validate the novel computer-aided design approach in this research.