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This study aimed to evaluate the value of cystatin C (Cys C) in predicting the perioperative and long-term prognosis of renal transplantation (RT). The clinical data of 198 RT recipients were collected. Blood samples were obtained daily until 7 d after transplantation and then discharge day to determine the serum levels of Cys C. The receiver-operating characteristic (ROC) analysis and the area under the curve (AUC) were used to determine the diagnostic accuracy of Cys C for delayed graft function (DGF). The presence of shrunken pore syndrome (SPS) with a cystatin C-based estimate of glomerular filtration rate less than 70% of a creatinine-based estimate, was also evaluated as a prognostic factor for the development of DGF. The serum Cys C levels of patients with DGF were higher than those of the non-DGF group. Cys C showed a higher AUC (0.928) in the ROC analysis than did sCr (0.862). Compared to the non-SPS group, there were more patients diagnosed with SPS in the DGF group (p < .05). The follow-up data showed that patients diagnosed with SPS had higher levels of sCr and Cys C compared to other patients, suggesting a poor long-term prognosis. Our findings suggest that Cys C is a sensitive indicator of renal function during the perioperative period. Cys C at a concentration of 4.9 mg/L had the highest sum of sensitivity and specificity for prediction of DGF, with a sensitivity of 0.889 and a specificity of 0.8. SPS is associated with the development of DGF and the poor long-term prognosis of RT.
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Cistatina C , Transplante de Rim , Biomarcadores , Creatinina , Função Retardada do Enxerto/diagnóstico , Taxa de Filtração Glomerular , Humanos , Transplante de Rim/efeitos adversos , Prognóstico , Curva ROCRESUMO
BACKGROUND The aim of this study was to assess the diagnostic utility of iron homeostasis determinations for prediction of severity of COVID-19. MATERIAL AND METHODS This was a retrospective study enrolling a total of 50 patients diagnosed with the novel coronavirus disease-19 (COVID-19) from February 27, 2020 to March 30, 2020, including a severe group (12 patients) and a mild group (38 patients). For the control group, 50 healthy people were examined during the same period. We compared clinical laboratory data and iron homeostasis biomarkers among the 3 groups. ROC curve analysis was used to assess diagnoses. RESULTS Patients diagnosed with severe COVID-19 had higher hepcidin and serum ferritin levels than in other groups (p<0.001). A combination test of hepcidin and serum ferritin provided the best specificity and sensitivity in the prognosis of COVID-19 severity. Logistic regression analysis showed hepcidin and serum ferritin independently contributed to the severity of COVID-19. Hepcidin and serum ferritin tandem testing predicted COVID-19 severity with 94.6% specificity, while hepcidin and serum ferritin parallel testing had a sensitivity of 95.7%. CONCLUSIONS Iron homeostasis had a robust association with the occurrence of severe COVID-19. Iron homeostasis determinations were specific and sensitive for the early prediction of disease severity in COVID-19 patients and thus have clinical utility.
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Betacoronavirus , Infecções por Coronavirus/sangue , Ferritinas/sangue , Hepcidinas/sangue , Pandemias , Pneumonia Viral/sangue , Adulto , Idoso , Área Sob a Curva , Biomarcadores , COVID-19 , Feminino , Homeostase , Humanos , Ferro/metabolismo , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Prognóstico , Curva ROC , Estudos Retrospectivos , SARS-CoV-2 , Sensibilidade e EspecificidadeRESUMO
Our present work was aimed to study on the regulatory role of MALAT1/miR-145-5p/AKAP12 axis on docetaxel (DTX) sensitivity of prostate cancer (PCa) cells. The microarray data (GSE33455) to identify differentially expressed lncRNAs and mRNAs in DTX-resistant PCa cell lines (DU-145-DTX and PC-3-DTX) was retrieved from the Gene Expression Omnibus (GEO) database. QRT-PCR analysis was performed to measure MALAT1 expression in DTX-sensitive and DTX-resistant tissues/cells. The human DTX-resistant cell lines DU145-PTX and PC3-DTX were established as in vitro cell models, and the expression of MALAT1, miR-145-5p and AKAP12 was manipulated in DTX-sensitive and DTX-resistant cells. Cell viability was examined using MTT assay and colony formation methods. Cell apoptosis was assessed by TUNEL staining. Cell migration and invasion was determined by scratch test (wound healing) and Transwell assay, respectively. Dual-luciferase assay was applied to analyse the target relationship between lncRNA MALAT1 and miR-145-5p, as well as between miR-145-5p and AKAP12. Tumour xenograft study was undertaken to confirm the correlation of MALAT1/miR-145-5p/AKAP12 axis and DTX sensitivity of PCa cells in vivo. In this study, we firstly notified that the MALAT1 expression levels were up-regulated in clinical DTX-resistant PCa samples. Overexpressed MALAT1 promoted cell proliferation, migration and invasion but decreased cell apoptosis rate of PCa cells in spite of DTX treatment. We identified miR-145-5p as a target of MALAT1. MiR-145-5p overexpression in PC3-DTX led to inhibited cell proliferation, migration and invasion as well as reduced chemoresistance to DTX, which was attenuated by MALAT1. Moreover, we determined that AKAP12 was a target of miR-145-5p, which significantly induced chemoresistance of PCa cells to DTX. Besides, it was proved that MALAT1 promoted tumour cell proliferation and enhanced DTX-chemoresistance in vivo. There was an lncRNA MALAT1/miR-145-5p/AKAP12 axis involved in DTX resistance of PCa cells and provided a new thought for PCa therapy.
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Proteínas de Ancoragem à Quinase A/genética , Proteínas de Ciclo Celular/genética , MicroRNAs/genética , Neoplasias da Próstata/tratamento farmacológico , RNA Longo não Codificante/genética , Animais , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Docetaxel/administração & dosagem , Resistencia a Medicamentos Antineoplásicos/genética , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Masculino , Camundongos , Próstata/efeitos dos fármacos , Próstata/patologia , Neoplasias da Próstata/genética , Neoplasias da Próstata/patologia , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
In prior research, evidence has been found for a relation between low exposure of long non-coding RNAs (lncRNAs) and prostate tumor genesis. This study aims to clarify the underlying mechanisms of lncRNA GAS5 in prostate cancer (PCa). In total, 118 pairs of PCa tissues and matched adjacent non-tumor tissues were collected. Additionally, lncRNA GAS5 exposure levels were determined using RT-PCR and in situ hybridization. In addition, dual-luciferase report assay was performed to verify the target effect of lncRNA GAS5 on miR-103. The exposure levels of the proteins related to the protein kinase B (AKT)/mammalian target of rapamycin (mTOR) axis, including AKT, mTOR, and S6K1, were measured by western blot PC3 cells infected with lncRNA GAS5 mimic; lncRNA GAS5 siRNA; or a combination of lncRNA and miR-103. The proliferation, invasion, and migration ability of PC3 cells after being infected were tested by MTT assay, wound healing assay, and transwell assays. Finally, nude mouse xenograft models were used to measure lncRNA GAS5 effects on prostate tumor growth in vivo. The lncRNA GAS5 levels were reduced significantly in the PCa tissues and cell lines (P < 0.05). A low exposure of lncRNA GAS5 caused AKT/mTOR signaling pathway activation in PC3 cells (P < 0.05). In addition, over-exposure of lncRNA GAS5 was proven to significantly decelerate PCa cell progression in vitro and tumor growth in vivo through inactivating the AKT/mTOR signaling pathway (P < 0.05). This study proves that lncRNA GAS5 plays a significant role in the decelerating PCa development via mediating the AKT/mTOR signaling pathway through targeting miR-103.
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BACKGROUND To explore the use of hepcidin as a marker of impaired renal function in a rat model for chronic allograft nephropathy (CAN). MATERIAL AND METHODS Twenty-four models were developed and 20 models were included in this study, using Fisher (F344) rats (donors) and Lewis rats (recipients). Renal function tests were performed preoperatively and postoperatively. Hepcidin, interleukin-6 (IL-6), and erythropoietin levels in serum and urine were measured by enzyme-linked immunosorbent assay (ELISA). To observe pathological changes in the kidneys, 10 rats each were sacrificed at 2 months and 4 months after surgery. RESULTS After transplantation, the serum hepcidin and IL-6 levels increased, while urine hepcidin levels decreased. Erythropoietin levels showed a similar trend; all P<0.05. Serum creatinine (SCr) and blood urea nitrogen significantly increased post-operatively, with SCr positively correlating with serum hepcidin. Serum hepcidin positively correlated with IL-6 and negatively correlated with EPO. Histopathological results were consistent with CAN, after transplantation. CONCLUSIONS Hepcidin may be considered as a potential marker of impaired renal function.
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Aloenxertos/metabolismo , Hepcidinas/metabolismo , Nefropatias/fisiopatologia , Testes de Função Renal , Transplante de Rim , Animais , Biomarcadores/sangue , Biomarcadores/metabolismo , Modelos Animais de Doenças , Hepcidinas/sangue , Nefropatias/sangue , Nefropatias/cirurgia , Ratos Endogâmicos F344 , Ratos Endogâmicos Lew , Resultado do TratamentoRESUMO
Prostate cancer and prostatic hyperplasia detection remains a great challenge, lacking of effective non-invasive and specific diagnostic biomarkers. In the current study, we aimed to identify the relative expression of plasma MD-miniRNA and its diagnostic performance in differentiating prostate cancer and prostatic hyperplasia patients from healthy controls, compared with serum prostate-specific antigen (PSA) level. All of the clinical participants (63 prostate cancer patients, 32 prostatic hyperplasia patients, and 50 healthy controls) were obtained from the Third Affiliated Hospital of Suzhou University in China between January 2013 and April 2014. Clinical characteristics were well matched. Plasma samples were extracted to test the relative expression of MD-miniRNA using the method of qRT-PCR. SPSS 22.0 statistical software package was used to analyze the data and GraphPad Prism 6.0 was used to generate the graphs. Relativity expression of plasma MD-miniRNA was significantly upregulated in prostate cancer, compared with prostatic hyperplasia patients and healthy controls. Serum PSA level revealed similar differences among these groups. MD-miniRNA presented a relatively high diagnostic accuracy with AUC of 0.86 (95 % CI 0.80-0.93) in differentiating prostate cancer patients from healthy controls. Simultaneously, MD-miniRNA was able to discriminate prostate cancer patients from prostatic hyperplasia controls with AUC of 0.79 (95 % CI 0.70-0.88). In addition, MD-miniRNA displayed a better diagnostic performance than PSA level. However, the panel of these two biomarkers revealed the best diagnostic performance, compared with either single biomarker. Results of this study showed that plasma MD-miniRNA could serve as a promising and noninvasive biomarker for diagnosing prostate cancer. Further large-scale studies are needed to confirm its clinical diagnosis accuracy.
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Antígeno Prostático Específico/sangue , Hiperplasia Prostática/sangue , Neoplasias da Próstata/sangue , RNA Longo não Codificante/sangue , Idoso , Diagnóstico Diferencial , Humanos , Masculino , Pessoa de Meia-Idade , Antígeno Prostático Específico/biossíntese , Hiperplasia Prostática/genética , Neoplasias da Próstata/genética , RNA Longo não Codificante/biossínteseRESUMO
OBJECTIVE: To investigate the expression of follistatin-like protein 1 (FSTL-1) in bone metastasis of prostate cancer (BMPC), the correlation of serum FSTL-1 with the chronic inflammatory factor interleukin-6 (IL-6) and bone morphogenetic protein 6 (BMP6) , and the clinical application value of serum FSTL-1 in BMPC. METHODS: Using ELISA, we measured the expression levels of serum FSTL-1, IL-6, and BMP6 in 35 patients with BMPC and another 30 with benign prostatic hyperplasia (BPH) and performed correlation analysis on the data obtained. RESULTS: Compared with the BPH controls, the BMPC patients showed a significantly decreased expression of serum FSTL-1 ([34.45 ± 12.35] µg/L vs [20.23 ± 8.69] µg/L, P < 0.01) and increased levels of IL-6 ([11.21 ± 8.62] µg/L vs [23.56 ± 20.12] µg/L, P < 0.05) and BMP6 ([293.50 ± 39.72] µg/L vs [428.30 ± 178.40] µg/L, P < 0.05). There was a significant negative correlation between the level of serum FSTL-1 and those of IL-6 and BMP6 in the BMPC patients, with correlation coefficients of -0.971 and -0.972, respectively (P < 0.05). CONCLUSION: The expression of serum FSTL-1 decreases in patients with bone metastasis of prostate cancer, and it is correlated with the levels of inflammatory factor and cell transformation factor. This finding offers a novel biological marker for the development and progression of prostate cancer as well as a new biological target factor for its intervention.
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Biomarcadores Tumorais/sangue , Neoplasias Ósseas/sangue , Neoplasias Ósseas/secundário , Proteínas Relacionadas à Folistatina/sangue , Interleucina-6/sangue , Hiperplasia Prostática/sangue , Neoplasias da Próstata/patologia , Idoso , Proteína Morfogenética Óssea 6/sangue , Progressão da Doença , Humanos , Masculino , Neoplasias da Próstata/sangueRESUMO
N6-methyladenosine (m6A) is the most prevalent reversible modification in eukaryotic mRNA, and it plays a critical role in tumor progression. The purpose of this study was to investigate the function and regulatory mechanisms of the methyltransferase METTL3 in renal cell carcinoma (RCC). METTL3 expression was upregulated and predicted a poor prognosis in patients with advanced RCC. METTL3 facilitated the proliferation, migration, and invasion of RCC cells, depending on its methylase activity. METTL3 positively regulated the expression of PLOD2, and both genes were triggered under prolonged hypoxia. Mechanistically, hypoxia-induced the binding of HIF-1α to the METTL3 promoter, which enhanced its transcriptional activity. METTL3-mediated m6A modifications of PLOD2 mRNA at 3'UTR region, promoting the translation of PLOD2 protein. Furthermore, silencing METTL3 impaired RCC progression in vitro. In vivo, administration of highly potent and selective METTL3 inhibitor STM2457 showed anti-tumor effects, whereas AAV9-mediated re-transduction of PLOD2 largely abolished the above phenomenon in a subcutaneous mouse model. These findings reveal that hypoxia and HIF-driven METTL3 transcription promote RCC progression by increasing PLOD2 expression in an m6A-dependent manner, suggesting that METTL3 may serve as a novel pharmaceutical intervention for RCC.
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Carcinoma de Células Renais , Neoplasias Renais , Camundongos , Animais , Humanos , Carcinoma de Células Renais/genética , Metiltransferases/metabolismo , Metilação , Neoplasias Renais/genética , Hipóxia/genética , RNA Mensageiro/genética , Pró-Colágeno-Lisina 2-Oxoglutarato 5-Dioxigenase/metabolismoRESUMO
Tumors of the male genitourinary system are of great concern to the health of men worldwide. Although emerging experiment-based evidence indicates an association between hepcidin and such cancers, an integrated analysis is still lacking. For this reason, in this study, we determined the underlying oncogenic functions of hepcidin in common male genitourinary system tumors, including bladder urothelial carcinoma (BLCA), kidney chromophobe (KICH), kidney renal clear cell carcinoma (KIRC), kidney renal papillary cell carcinoma (KIRP), prostate adenocarcinoma (PRAD), and testicular germ cell tumors (TGCT) according to the data from The Cancer Genome Atlas. We found that hepcidin was highly expressed in kidney and testicular cancers. Meanwhile, the expression level of hepcidin was distinctly associated with the prognosis and immune cell infiltration in male patients with certain genitourinary system cancers, especially in KIRC. Elevated hepcidin levels also present as a risk factor in male genitourinary system tumors. Moreover, enrichment analyses revealed that some of the principal associated signaling pathways involving hepcidin and its related genes are identified as tumorigenesis-related. Immunofluorescence staining confirmed the conclusion of our immune infiltration analysis in KIRC tissue. In this study, for the first time, we provided evidence for the oncogenic function of hepcidin in different types of male genitourinary system tumors.
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Short-chain fatty acids (SCFAs), the end products of fermentation carried out by the intestinal microbiota, were demonstrated to produce anti-oxidant and anti-inflammatory effects. Butyrate, part of the SCFAs, also shows the same effect. Renal ischemia/reperfusion (I/R) injury commonly occurs in renal transplantation and is often accompanied by oxidative stresses and inflammatory responses. In this study, we explore butyrate effect on renal I/R injury and SCFAs changes in renal transplant. Male Sprague-Dawley rats were pretreated with butyrate as research, and underwent the surgery of renal ischemia for 45 min followed by reperfusion. 90 rats were randomly divided into 3 groups (n=30 each group): (1) sham-operated group; (2) butyrate-treated group; (3) control group. The samples of blood and renal were collected immediately for further studies. Thirty-two patients were enrolled to investigate the levels of SCFAs after the renal transplantation. Rats model showed that butyrate treatments significantly enhanced the function and structure of kidney, as evidenced by the lower serum creatinine levels and less pathological damages of renal tissue. With the recovery of renal function after renal transplantation, SCFAs increased, which were negatively correlated with creatinine. Butyrate expressed like SCFAs. In this study, we demonstrated that butyrate increased with the recovery of renal function after renal transplantation. Most importantly, butyrate treatments alleviated the renal damages caused by I/R via the upregulation of intracellular oxidant stress and inflammations.
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Butiratos/farmacologia , Traumatismo por Reperfusão/prevenção & controle , Injúria Renal Aguda/cirurgia , Animais , Butiratos/uso terapêutico , Creatinina , Ácidos Graxos , Ácidos Graxos Voláteis , Humanos , Rim/fisiologia , Transplante de Rim , Masculino , Ratos , Ratos Sprague-Dawley , Traumatismo por Reperfusão/tratamento farmacológicoRESUMO
BACKGROUND: It has been widely recognized that exosomal miRNAs can participate in the pathogenesis of different renal disorders and serve as disease biomarkers. Although kidney biopsy is still the gold standard for diagnosing and monitoring immunoglobulin A nephropathy (IgAN), it is highly required to identify new and effective noninvasive biomarkers for IgAN, the most frequently detected primary glomerulonephritis worldwide. METHODS: Plasma and urinary exosomes were extracted by PEG precipitation. Size and morphological characteristics of plasma and urinary exosomes were observed by transmission electron microscopy and nanoparticle tracking analysis. The levels of plasma and urinary exosomes were revealed by Western blotting. The expressions of target miRNAs were revealed by in situ hybridization and qRT-PCR. RESULTS: The levels of plasma and urinary exosomes were remarkably enhanced in IgAN patients compared with healthy controls (HCs). The expressions of miR-4639 and miR-210 in IgAN patients were significantly higher in contrast to the individuals with membranous nephropathy, minimal change nephrosis, diabetic nephropathy, or HC. These also played a valuable role in assessing the kidney function and the level of proteinuria. Furthermore, plasma and urinary exosomal miR-4639 expression was associated with more serious and active histological activity (mesangial hypercellularity, crescent, and C3 complement deposition). With an average follow-up of 8 months, miR-4639 and miR-210 expressions in plasma and urinary exosomes were higher in patients with progressive IgAN. Plasma exosomal miR-4639 and miR-210 were better than proteinuria (g/24 h) to estimate renal outcomes. CONCLUSION: Exosomal miR-4639 and miR-210 could be used as valid biomarkers to assist in diagnosis, evaluate severity, and assess disease development of IgAN.
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Glomerulonefrite por IGA , MicroRNAs , Humanos , Glomerulonefrite por IGA/diagnósticoRESUMO
[This corrects the article DOI: 10.3892/ol.2015.3363.].
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Prostate cancer (PCa) is the most commonly diagnosed cancer in males and the fifth most common cause of cancer death worldwide. Previous studies indicated that miR-18a-5p modulated epithelial-mesenchymal transition in breast cancer via targeting SREBP1 forming a co-repressor complex with Snail and HDAC1/2. However, the function of miR-18a-5p in prostate cancer remains largely unknown. In this study, we identified miR-18a-5p as a tumor promoter in prostate cancer. miR-18a-5p expression was found upregulated in human prostate cancer tissues while SLC40A1 was down-regulated. Cell proliferation assay demonstrated that miR-18a-5p promoted prostate cancer cell proliferation. We also found SLC40A1 was downregulated by miR-18a-5p in prostate cancer cell lines. Restoration of SLC40A1 reversed the effects of miR-18a-5p in prostate cancer cells. Taken together, our results suggest that miR-18a-5p might function as a tumor-promoting factor in PCa and might contribute to its proliferation.
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Proliferação de Células/genética , Transição Epitelial-Mesenquimal , Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , Neoplasias da Próstata/genética , Regulação para Cima , Linhagem Celular Tumoral , Humanos , Masculino , MicroRNAs/metabolismo , Neoplasias da Próstata/metabolismo , Neoplasias da Próstata/patologiaRESUMO
The purpose of the present study was to investigate the expressions of hsa-let-7c-5p and TGF-ß signaling-related molecules and their correlations with clinical characteristics in chronic kidney disease (CKD). Twenty-three biopsy specimens of CKD patients and 20 negative control tissues were selected. Quantitative real-time PCR (qPCR) was used for the detection of hsa-let-7c-5p, transforming growth factor ß (TGF-ß) and TGF-ß receptor type 1 (TGF-ßR1) expression levels. Target gene of hsa-let-7c-5p was verified by dual-luciferase reporter assay. A significant decrease of hsa-let-7c-5p expression in CKD tissue was found, compared with that of normal renal tissues (p < 0.01). Expression levels of TGF-ß in CKD were increased, compared with that of normal kidney tissue (p < 0.001). The difference in the expression of TGF-ß R1 between CKD tissues and normal renal tissues was not significant (p > 0.05). A negative correlation was found between the expression of TGF-ß and renal tissue hsa-let-7c-5p levels. Furthermore, hsa-let-7c-5p was identified to regulate TGF- ß1 by directly binding with the 167-173 site in the 3' untranslated region. Decreased hsa-let-7c-5p levels in CKD patients was found to be associated with disease severity, which shows a negative correlation with proteinuria and creatinine levels, and a positive correlation with estimated glomerular filtration rate (eGFR), while relative TGF-ß1 expression had a positive correlation with creatinine level. In summary, changes in hsa-let-7c-5p expression and its target gene TGF-ß are associated with the disease status of CKD. Let-7c-5p may contribute to the pathogenesis of renal fibrosis through TGF-ß signaling, a potential diagnostic and therapeutic target of the disease.
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MicroRNAs/metabolismo , Insuficiência Renal Crônica/metabolismo , Fator de Crescimento Transformador beta/metabolismo , Adulto , Feminino , Humanos , Rim/metabolismo , Rim/patologia , Masculino , MicroRNAs/genética , Pessoa de Meia-Idade , Insuficiência Renal Crônica/genética , Insuficiência Renal Crônica/patologia , Transdução de Sinais/genética , Fator de Crescimento Transformador beta/genéticaRESUMO
Aim: To systematically profile the global m6A modification pattern in clear cell renal cell carcinoma (ccRCC). Methods: m6A modification patterns in ccRCC and normal tissues were described via m6A sequencing and RNA sequencing, followed by bioinformatics analysis. m6A-related RNAs were immunoprecipitated and validated by quantitative real-time PCR (qPCR). Results: In total, 6919 new m6A peaks appeared with the disappearance of 5020 peaks in ccRCC samples. The unique m6A-related genes in ccRCC were associated with cancer-related pathways. We identified differentially expressed mRNA transcripts with hyper-methylated or hypo-methylated m6A peaks in ccRCC. Conclusion: This study presented the first m6A transcriptome-wide map of human ccRCC, which may shed lights on possible mechanisms of m6A-mediated gene expression regulation.
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Adenosina/análogos & derivados , Carcinoma de Células Renais/genética , Regulação Neoplásica da Expressão Gênica , Neoplasias Renais/genética , Adenosina/metabolismo , Carcinoma de Células Renais/metabolismo , Humanos , Neoplasias Renais/metabolismo , Proteínas de Ligação a RNA/metabolismo , RNA-Seq , Transdução de Sinais , TranscriptomaRESUMO
OBJECTIVE: The microRNA expression profile of plasma exosomes in prostate cancer (PCa) is of critical importance in the disease exploration. This study aimed to explore the clinical application of exosomal miRNAs as biomarkers for PCa. METHODS: Exosome-like vesicles of PCa patients and healthy controls were purified by differential centrifugation. The purified vesicles within the ranges of 50 and 100 nm were classified as exosomes according to the results of transmission electron microscopy and Western blot. Both, in vitro and in vivo, validations were performed by small RNA sequencing, CCK8, RT-qPCR, flow cytometry, Western blot, transwell and immunofluorescent staining assays. RESULTS: High-throughput sequencing identified that 94 miRNAs were differentially expressed in PCa patients in comparison with healthy controls (P<0.01; fold change ≥2). Among them, 64 miRNAs were upregulated, and 30 miRNAs were downregulated. In comparison to the healthy controls, the expression levels of miR-217 were significantly upregulated, while miR-23b-3p were significantly downregulated in the exosomes and serum collected from PCa patients. Both, in vitro and in vivo, studies revealed that exosomes secreted by PCa cells with up-regulated miR-217 levels promoted cell proliferation and invasion; meanwhile, the exosomes with up-regulated miR-23b-3p levels inhibited cell proliferation and invasion. The epithelial-mesenchymal transition process may have been involved in the above-mentioned regulation. CONCLUSION: This study identified the dysregulated expression of exosomal miRNAs in PCa patients, including miR-217 and miR-23b-3p, by validating their function on proliferation and invasion in PCa cells. This regulation may have been affected by the epithelial-mesenchymal transition process, suggesting that they can be used as potential targets in the diagnosis and treatment of PCa.
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BACKGROUND: Bladder cancer (BC) is a common type of cancer that involves tumors of the urinary system and poses a serious threat to human health. Long noncoding RNAs (lncRNAs) have emerged as crucial biomarkers and regulators in many cancers. Novel lncRNA biomarkers in BC urgently need to be investigated in regard to its function and regulatory mechanisms. METHODS: Identification of differentially expressed lncRNAs in BC tissue was performed via microarray analysis. To investigate the biological functions of LINC00612, loss-of-function and gain-of-function experiments were performed in vitro and in vivo. Bioinformatics analysis, dual-luciferase reporter assays, AGO2-RIP assays, RNA pull-down assays, real-time quantitative PCR (RT-qPCR) arrays, fluorescence in situ hybridization assays, and western blot assays were conducted to explore the underlying mechanisms of competitive endogenous RNAs (ceRNAs). RESULTS: LINC00612 was upregulated in BC tissues and cell lines. Functionally, downregulation of LINC00612 inhibited cell proliferation and invasion in vitro and in vivo, whereas overexpression of LINC00612 resulted in the opposite effects. Bioinformatics analysis and luciferase assays revealed that miR-590 was a direct target of LINC0061, which was validated by dual-luciferase reporter assays, AGO2-RIP assays, RNA pull-down assays, RT-qPCR arrays, and rescue experiments. Additionally, miR-590 was shown to directly target the PHD finger protein 14 (PHF14) gene. LNIC00612 modulated the expression of E-cadherin and vimentin by competitively sponging miR-590 to elevate the expression of PHF14, thus affecting BC cellular epithelial-mesenchymal transition (EMT). CONCLUSIONS: Our results indicate that LINC00612 enhances the proliferation and invasion ability of BC cells by sponging miR-590 to upregulate PHF14 expression and promote BC cellular EMT, suggesting that LINC00612 may act as a potential biomarker and therapeutic target for BC.
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MicroRNAs/genética , Proteínas Nucleares/genética , RNA Longo não Codificante/genética , Fatores de Transcrição/genética , Neoplasias da Bexiga Urinária/patologia , Animais , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Regulação Neoplásica da Expressão Gênica , Humanos , Camundongos , Invasividade Neoplásica , Neoplasias da Bexiga Urinária/genéticaAssuntos
Peptídeos Catiônicos Antimicrobianos/genética , Expressão Gênica , Rim/metabolismo , Traumatismo por Reperfusão/genética , Animais , Peptídeos Catiônicos Antimicrobianos/sangue , Creatinina/sangue , Hepcidinas , Interleucina-6/sangue , Rim/irrigação sanguínea , Ratos , Ratos Sprague-Dawley , Traumatismo por Reperfusão/sangue , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de TempoRESUMO
OBJECTIVE: This study investigated the prognostic factors and outcomes of a large observational cohort of patients with primary transitional cell carcinoma of the ureter, which was obtained from the Surveillance, Epidemiology, and End Results database. METHODS: We used the Surveillance, Epidemiology, and End Results program to identify 1910 patients who had available clinical and follow-up information and were diagnosed for primary transitional cell carcinoma of the ureter between 2004 and 2013. Descriptive statistics were used to explore the epidemiology, treatment practices, and tumor characteristics of the patients. Univariate and multivariable Cox regression models were used to analyze the patient data. RESULTS: The median overall survival (OS) was 46 months, and the 5-year OS rate was 41.8%. The median CSS was 78 months, and the 5-year CSS rate was 54.3%. Multivariate analysis identified tumor grade, tumor size, AJCC stage, M stage, cancer-directed surgical procedure and radiation as independent factors of primary transitional cell carcinoma of the ureter. For early stage patients, the surgical procedure was associated with fairly longer survival and additional radiation may cause more harm than benefit. Meanwhile, for advanced stage patients, the impact of surgery on OS and CSS greatly decreased. Radiation exerted a very limited impact on clinical outcomes. Patients with bad tumor differentiation or a large tumor size were more likely to have advanced stage disease. CONCLUSION: Durable cancer control can be expected in patients treated with surgery for early stage UTUC. The presence of advanced stage disease exerts a profound detrimental effect on the survival of patients.