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ABSTRACT Purpose: The regulatory effect of microRNA on diseases has been confirmed. This study aimed to evaluate the expression of microRNA-210-3p in age-related cataracts and assess the effect of abnormal miR-210-3p expressions on H2O2-induced SAR01/04 cells. Methods: Reverse-transcription quantitative polymerase chain reaction method was performed to assess the levels of miR-210-3p in aqueous humor samples. Receiver operating characteristic analysis was employed to assess the discrimination ability of miR-210-3p between patients with age-related cataracts and healthy people, and Pearson correlation analysis was used to identify the correlation between miR-210-3p and oxidative stress indices such as superoxide dismutase, glutathione peroxidase, malonaldehyde. Cell counting kit-8 assay and Transwell assay were used to estimate the biological function of H2O2-induced age-related cataract cell model. The levels of oxidative stress indices such as superoxide dismutase, glutathione peroxidase, and malonaldehyde were measured to evaluate the degree of oxidative stress damage in the age-related cataract cell model. The relationship between miR-210-3p and its target gene was verified by luciferase reporter gene analysis. Results: The miR-210-3p expression was elevated in the aqueous humor of patients with age-related cataracts. A high miR-210-3p expression showed a high diagnostic value for age-related cataracts and was significantly associated with the level of oxidative stress markers in patients with age-related cataracts. The inhibition of miR-210-3p can reverse oxidative stress stimulation and adverse effects on H2O2-induced cell function. Conclusions: The results suggested that miR-210-3p could promote cell viability, cell migration, and oxidative stress by targeting autophagy-related gene 7 in in vitro age-related cataract cell model.
RESUMO Objetivo: O efeito regulador do microRNA em doenças tem sido confirmado, e este artigo tentou avaliar a expressão do microRNA-210-3p na catarata relacionada à idade e avaliar o efeito da expressão anormal do miR-210-3p em células SAR01/04 induzidas por H2O2. Métodos: O método de transcrição reversa seguida de reação em cadeia da polimerase (RT-PCR) quantitativa foi realizado para avaliar os níveis de miR-210-3p em amostras de humor aquoso. Análise de características operacionais do receptor foi feita para avaliar a capacidade de discriminação do miR-210-3p entre pacientes com catarata relacionada à idade e pessoas saudáveis. A análise de correlação de Pearson identificou a correlação do miR-210-3p e índices de estresse oxidativo, como superóxido dismutase, glutationa peroxidase, malonaldeído. O ensaio de contagem de células kit-8 (cck-8) e o ensaio no sistema Transwell foram utilizados para estimar a função biológica do formato de células de catarata relacionada com a idade induzida por H2O2. Os níveis de índices de estresse oxidativo como superóxido dismutase, glutationa peroxidase e malonaldeído foram detectados para avaliar o grau de dano do estresse oxidativo em formato de células de catarata relacionada à idade. A relação entre miR-210-3p e seu gene alvo foi verificada por análise do gene repórter luciferase. Resultados: A expressão miR-210-3p foi elevada no humor aquoso de pacientes com catarata relacionada à idade. A expressão miR-210-3p altamente expressiva mostrou alto valor diagnóstico para catarata relacionada à idade e foi significativamente associado ao nível de marcadores de estresse oxidativo em pacientes com catarata relacionada à idade. A inibição de miR-210-3p pode reverter a estimulação do estresse oxidativo e os efeitos adversos da função celular induzida por H2O2. Conclusões: Esses dados sugeriram que a expressão miR-210-3p poderia promover a viabilidade celular, migração celular e estresse oxidativo ao direcionar genes ATG 7 relacionados à autofagia em modelo in vitro de células de catarata relacionadas à idade.
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The aim of this study was to investigate the role of seminal plasma miR-210-3p in the impairment of semen quality caused by varicocele. This study included 102 patients whose semen quality was normal when they were diagnosed with varicocele. A 2-year follow-up for included patients was performed, and they were divided into Group A (semen quality became abnormal) and Group B (semen quality remained normal) according to the results of semen analysis during the follow-up. Semen parameters and seminal plasma miR-210-3p expression were investigated by semen analysis and quantitative real-time polymerase chain reaction, respectively. In vitro experiments with GC-2 cells were performed to explore the role of miR-210-3p in spermatogenic cells. The results of quantitative real-time polymerase chain reaction showed that the level of seminal plasma miR-210-3p in Group A was higher than that in Group B both after 2-year follow-up and when they were diagnosed with varicocele (both P < 0.01). Apoptosis and proliferation assays showed that miR-210-3p induces apoptosis of spermatogenic cells by promoting caspase-3 activation. In conclusion, our study indicated that seminal plasma miR-210-3p induces spermatogenic cell apoptosis by activating caspase-3 in patients with varicocele. Seminal plasma miR-210-3p may be a potential biomarker for predicting impaired semen quality caused by varicocele.
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BACKGROUND: Porous hydroxyapatite scaffolds have good osteogenesis in vivo and in vitro. However, little research has been done on the complex regulation mechanisms of miRNAs involved. OBJECTIVE: To investigate the changes of related miRNA expression in rat bone marrow mesenchymal stem cells during osteogenic mineralization by porous hydroxyapatite scaffolds. METHODS: Rat bone marrow mesenchymal stem cells were isolated, cultured and identified in vitro. Bone marrow mesenchymal stem cells co-cultured with porous hydroxyapatite scaffold were as experimental group, and bone marrow mesenchymal stem cells cultured alone served as blank control group, both of which underwent osteogenic induction for 7 days. During the osteogenic mineralization, miRNA high-throughput sequencing technology was used to analyze the changes of miRNA expression profiles followed by GO analysis. The miRNA molecules with obvious expression differences were screened and verified by qRT-PCR. RESULTS AND CONCLUSION: (1) Compared with the blank control group, in the experimental group, the expression levels of BMP2, ALP and Runx2 mRNA were up-regulated, and the expression level of BMP2 was up-regulated significantly (P < 0.05). (2) Results of miRNA high-throughput sequencing showed that 13 miRNAs such as miR-210-3p and miR-146a-5p were up-regulated, and 17 miRNAs such as let-7c-3p and let-3615 were down-regulated significantly. (3) GO analysis revealed that up-regulated miRNA target genes were mainly involved in biological regulation, cellular gene expression, and gene expression regulation, mainly including nuclear factor-κB, Toll-like receptor 9, intercellular adhesion, interleukin-1 regulation, and signaling pathways such as angiogenesis and Hippo. (4) Real-time fluorescence quantitative qPCR results showed that miRNA-210 was up-regulated 15 times and miR-146a-5p was up-regulated 10 times in the experimental group (P < 0.05). These results indicate that the new microchannel porous hydroxyapatite scaffold can promote the differentiation of bone marrow mesenchymal stem cells by up-regulating miRNA-210-3p and miR-146a.
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Hepatocellular carcinoma (HCC) is one of the most common types of liver cancer and is the second leading cause of cancer mortality with an estimated 745 500 deaths annually (Jemal et al., 2011). Although new therapeutic modalities including novel chemotherapeutic interventions and targeted therapy have been applied, the prognosis of HCC patients remains unsatisfactory due to the high incidence of intrahepatic and distal metastases (Siegel et al., 2018).
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Female , Humans , Male , Apoptosis Regulatory Proteins/physiology , Biomarkers , Carcinoma, Hepatocellular/pathology , Genome , Hypoxia , Liver Neoplasms/pathology , MicroRNAs/analysis , Neoplasm Staging , Prognosis , Repressor Proteins/physiologyABSTRACT
Objective To investigate whether miR-210 plays a crucial role in the process of inducing the transfor-mation from co-cultured adipose derived(AD)-MSCs into CAFs. Methods Co-cultured adipose derived (AD)-MSCs with breast cancer cells (BCCs) lines in vitro and detected the expression of miR-210 using RT-qPCR. The expression of CAFs characteristic markers including α-SMA and tenascin-c were measured by RT-qPCR and the ex-pression of α-SMA and FAPA were assessed by Western blot.Co-injected MCF-7 cells transfected with miR-210 in-hibitor or miR-NC with MSC at 1 : 1 ratio into the immunodeficient nude mice subcutaneously and observed tumor growth in vivo constantly. Results miR-210 was all up-regulated when a variety of breast cancer cells were co-cul-tured with MSCs (P<0.05). Inhibition of miR-210 in tumor cell could inhibit the transformation from MSCs into CAFs(P<0.05). Animal experiments further confirmed that inhibition of miR-210 in tumor cell could reduce tumor growth (P<0.05). Conclusions As an important information molecule, miR-210 mediates the transforma-tion from MSCs to CAFs in the tumor microenvironment.
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Objective To investigate the expression and significance of serum thymidine kinase-1(TK1), soluble intercellular adhesion molecule-1(sICAM-1),miR-210,and carcinoembryonic antigen(CEA)in pa-tients with colorectal cancer.Methods A total of 200 patients with colorectal cancer treated in the hospital from 2015 to 2017 were enrolled as the observation group and 50 healthy subjects were enrolled as the control group.Serum levels of TK1,sICAM-1,miR-210,and CEA were measured before and after treatment,and the trend of each indicator was analyzed.To analyze the relationship between tumor site,degree of tumor differen-tiation,clinical stage,w hether it is the first patient,lymph node metastasis and miR-210 levels in patients with colorectal cancer.Results The concentrations of sICAM-1,CEA,sTK1 and miR-210 in the observation group were significantly higher than those in the control group(P<0.05).The expression of miR-210 was related to the clinical stage and lymph node metastasis(P<0.05).T he sensitivity,specificity,positive predictive value, negative predictive value and accuracy of the combined tests including serum TK 1,sICAM-1,miR-210 and CEA test were higher than those of the single test,and was also higher than the combined tests of TK1,miR-210 and CEA.The sensitivity of the four combined tests was 75.70%,the specificity was 86.00%,the positive predictive value was 82.00%,the negative predictive value was 88.00%,the accuracy was 92.40%.Conclusion The combined detection of serum TK1,sICAM-1,miR-210 and CEA may have some value in the early diag-nosis of colorectal cancer,and can improve the sensitivity and specificity of colorectal cancer diagnosis.
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Abstract Background: Crocin is reported to have a wide range of biological activities such as cardiovascular protection. Recent epidemiologic studies have shown that exercise reduces cardiovascular morbidity and mortality in the general population. Objective: The aim of this study was to evaluate the effect of crocin and voluntary exercise on miR-126 and miR-210 expression levels and angiogenesis in the heart tissue. Methods: Animals were divided into 4 groups: control, exercise, crocin, and exercise-crocin. Animals received oral administration of crocin (50 mg/kg) or performed voluntary exercise alone or together for 8 weeks. Akt, ERK1/2 protein levels, miR-126 and miR-210 expression were measured in the heart tissue. Immunohistochemical method was used to detect CD31 in the heart tissue. Results: Akt and ERK1/2 levels of the heart tissue were higher in crocin treated group and voluntary exercise trained group after 8 weeks. Combination of crocin and exercise also significantly enhanced Akt and ERK1/2 levels in the heart tissue. MiR-126, miR-210 expression and CD31 in the heart increased in both crocin and voluntary exercise groups compared with control group. In addition, combination of exercise and crocin amplified their effect on miR-126 and miR-210 expression, and angiogenesis. Conclusion: Crocin and voluntary exercise improve heart angiogenesis possibly through enhancement of miR-126 and miR-210 expression. Voluntary exercise and diet supplementation with crocin could have beneficial effects in prevention of cardiovascular disease.
Resumo Fundamentos: A crocina tem uma vasta gama de atividades biológicas, tais como a proteção cardiovascular. Estudos epidemiológicos recentes demonstraram que o exercício reduz a morbidade e a mortalidade cardiovasculares na população em geral. Objetivo: O objetivo deste estudo foi avaliar o efeito da crocina e do exercício voluntário nos níveis de expressão miR-126 e miR-210 e na angiogênese no tecido cardíaco. Métodos: Os animais foram divididos em 4 grupos: controle, exercício, crocina e exercício-crocina. Os animais receberam a administração oral de crocina (50 mg/kg) ou realizaram exercício voluntário sozinhos ou em conjunto durante 8 semanas. Os níveis de proteína Akt, ERK1/2, e a expressão de miR-126 e miR-210 foram medidos no tecido cardíaco. O método imunohistoquímico foi utilizado para detectar CD31 no tecido cardíaco. Resultados: Os níveis de Akt e ERK1/2 do tecido cardíaco foram maiores no grupo tratado com crocina e no grupo de exercício voluntário após 8 semanas. A combinação de crocina e exercício também aumentou significativamente os níveis de Akt e ERK1/2 no tecido cardíaco. A expressão de MiR-126, miR-210 e CD31 no coração aumentou tanto em no grupo de crocina como no grupo de exercício voluntário em comparação com o grupo de controle. Além disso, a combinação de exercício e crocina amplificou seu efeito na expressão de miR-126 e miR-210 e angiogênese. Conclusão: A Crocina e o exercício voluntário melhoram a angiogênese cardíaca possivelmente através do aumento da expressão de miR-126 e miR-210. O exercício voluntário e a suplementação dietética com crocina podem ter efeitos benéficos na prevenção de doenças cardiovasculares.
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Animals , Male , Rats , Physical Conditioning, Animal , Carotenoids/pharmacology , Neovascularization, Physiologic/physiology , MicroRNAs/metabolism , Diabetes Mellitus, Experimental/metabolism , Myocardium/metabolism , Time Factors , Immunohistochemistry , Rats, Wistar , MAP Kinase Signaling SystemABSTRACT
Objective:To investigate the effect of miR-210 on the migration and proliferation of lung cancer cells. Methods:The expression of miR-210 in normal lung tissues and lung cancer tissues were detected by qPCR. The expression of Mex3B in lung cancer tissues and adjacent tissues were detected by qPCR. qPCR was used to detect the expression of miR-210 in different lung cancer cell lines(A549,H1299,H1650 and H358). The effect of miR-210 on the transcription of Mex3B was detected by double luciferase reporter gene system. The effect of miR-210 expression on the activity of lung cancer cells was detected by cell viability assay. The effect of miR-210 on the proliferation of A549 cells were detected by plate cloning assay. Transwell invasion assay were used to detect the effect of miR-210 on the invasion ability of lung cancer cell line A549. Results:Compared with adjacent tissues,the expression of miR-210 were significantly decreased in lung cancer tissues,Mex3B were lower in lung cancer. Double luciferase reporter gene system results showed that miR-210 can directly regulate the transcriptional activity of Mex3B. The invasion and proliferation ability of lung cancer cell line were significantly reduced after inhibition of miR-210. Conclusion:miR-210 can target regulate the expression of Mex3B, and affects the invasion and proliferation of lung cancer cells.
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Objective To investigate the effect of miR-210 on angiogenesis of human renal clear cell carcinoma . Methods Detect the expression of miR-210 in 40 cases of renal clear cell carcinoma tissues , and analyze the rela-tionship between the expression of miR-210 and microvessel density ( MVD) .miR-210-ASO was lipotransfected in-to renal clear cell carcinoma cell line 786-O.RT-qPCR was used to verify the transfection effect .The effect of the supernatant of control group , negative control group and miR-210-ASO group tumor cells on the lumen formation of human umbilical vein endothelial cells ( HUVEC) was observed in a 3-D culturesystems .The transplantation tumor model of nude mice was established , and the effect of miR-210 on the formation of the transplanted tumor micro vessel was observed by endomucin and VEGF immunofluorescence staining under laser scanning confocal micro -scope.Results The expression of miR-210 was positively correlated with microvessel density in renal clear cell carcinoma ( P<0.05 ) .The expression of miR-210 was significantly decreased in 786-O cells transfected with miR-210-ASO( P<0.05 ) .The lumen formation of HUVEC cells co cultured with miR-210-ASO group cell supernatant was significantly less than that of control group and negative control group ( P<0.05 ) .The tumor volume of miR-210-ASO group was less than that of the control group , and the number of the micro vessel and the VEGF expres-sion were significantly less than that of the control group ( P<0.05 ) .Conclusions Inhibition of miR-210 can suppress blood vessel formation in renal clear cell carcinoma .
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Objective To explore the effect of anoxic microenvironment and hypoxia related microRNA-210 (miR-210) on the epithelial-mesenchymal transition (EMT) in gastric cancer cell line 803.Methods The mRNA relative expressions of E-cadherin and Twist1 were detected by qRT-PCR in gastric cancer cell line after being induced hypoxia by CoCl2 (100,200 and 300 μ mol/L),which was compared with normoxic group (0 mol/L).The cell proliferation of gastric cancer cell line was observed by cell proliferation assay.The relative expressions of E-cadherin and Twist 1 were detected in gastric cancer cell line after transfecting miR-210 mimics.Results With the increase of the concentrations of CoCl2,the relative expression of miR-210 and the proliferation rate of gastric cancer cells showed a tendency of increase first and then decrease (P ＜ 0.05).The relative expression of E-cadherin mRNA was lower in hypoxia group than that of normoxic group.The relative expression of Twist1 mRNA was higher in hypoxia group than that of normoxic group.The relative expressions of Ecadherin mRNA were significantly higher in 200 and 300 μ mol/L CoCl2 groups than that of 100 μmol/L CoCl2 group (P ＜0.05).There was no significant difference in the relative expression of Twist1 mRNA between the three CoCl2 groups.The relative expression of Twist1 mRNA was higher in the transfection group than that of control group.The relative expression of E-cadherin mRNA was significantly lower in the transfection group than that of control group (P ＜ 0.05).Conclusion Hypoxia can promote the proliferation and EMT of gastric cancer cells.The up-regulation of miR-210 can also promote the procession of EMT,which may be the intermediate link in the process of EMT induced by hypoxia.
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Objective:To analyze the expression levels of the HIF-1αin the tissues of gastric cancer and the molecular mechanism in the pathogenesis of the gastric cancer .Methods:A total of 60 patients resided at the Hehuang valley in Qinghai province and diagnosed to gastric cancer were collected to plateau group .And the 60 patients resided at the plains (Shenzhen) and diagnosed to gastric cancer were collected to plains group .The specimens of gastric cancer in two groups were collected .The levels of HIF-1αprotein were detected by the SP method of the immunological histochemistry and compared between two groups .The levels of HIF-1αgene were detected by the RT-PCR and compared between two groups .And the levels of miR-210 were detected by the RT-PCR and compared between two groups .To analyze the relationship between the expression of HIF-1αand clinicopathological features in patients with gastric cancer.Results:The levels of HIF-1αprotein in plateau group were higher than in plains group (P<0.05).The levels of HIF-1αgene in plateau group were higher than in plains group (P<0.05).The levels of miR-210 in plateau group were higher than in plains group(P<0.05).The positive expression of HIF-1αin gastric cancer was related to tumor differentiation (P=0.0441),invasion depth (P=0.0319),lymph node metastasis(P=0.0253) and TNM staging(P=0.0289).Conclusion:The expression levels of the HIF-1αin the tissues of gastric cancer are upregulated under hypoxia environment , and the HIF-1αcan improve the development of the gastric cancer.
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Objective:To investigate the effect of miR-210 on the migration and proliferation of lung cancer cells. Methods:The expression of miR-210 in normal lung tissues and lung cancer tissues were detected by qPCR. The expression of Mex3B in lung cancer tissues and adjacent tissues were detected by qPCR. qPCR was used to detect the expression of miR-210 in different lung cancer cell lines(A549,H1299,H1650 and H358). The effect of miR-210 on the transcription of Mex3B was detected by double luciferase reporter gene system. The effect of miR-210 expression on the activity of lung cancer cells was detected by cell viability assay. The effect of miR-210 on the proliferation of A549 cells were detected by plate cloning assay. Transwell invasion assay were used to detect the effect of miR-210 on the invasion ability of lung cancer cell line A549. Results:Compared with adjacent tissues,the expression of miR-210 were significantly decreased in lung cancer tissues,Mex3B were lower in lung cancer. Double luciferase reporter gene system results showed that miR-210 can directly regulate the transcriptional activity of Mex3B. The invasion and proliferation ability of lung cancer cell line were significantly reduced after inhibition of miR-210. Conclusion:miR-210 can target regulate the expression of Mex3B, and affects the invasion and proliferation of lung cancer cells.
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Objective To construct the lentiviral vector Lenti-miR-210-Luciferase, and to detect angiogenic factors HIF-1α and VEGF expression in hPDLSCs transduced by Lenti-miR-210-Luciferase. Methods hPDLSCs were iso-lated and cultured, and according to human miR-210 gene sequence(NC_000011. 9), its primer was designed and amplified through PCR. The PCR products of the target gene were connected to the vector pLVX-EGFP-3FLAG-EF1-Luc. To identify the plasmid, target gene PCR product and the purpose vector were digested by EcoRⅠand XbaⅠ. Lenti-miR-210-Luciferase ( the control group was Lenti-LacZ-Luciferase) was constructed using the LR re-combination system. After hPDLSCs was transduced by Lenti-miR-210-Luciferase, the analysis of HIF-1α and VEGF expression was done with qPCR and the immunohistochemistry examination. Results The results of plasmid sequencing and digestion confirmed that the vector Lenti-miR-210-Luciferase was successfully constructed. After Lenti-miR-210-Luciferase was transduced to hPDLSCs on 0, 1, 4, 7 and 14 d, the results of qPCR showed that the over-expression of HIF-1αand VEGF was detected on 4 d, and continued until 21 d. Immunohistochemical results showed that after hPDLSCs were transduced by Lenti-miR-210-Luciferase, hPDLSCs were positive for HIF-1α and VEGF antibody, and the control group was negative. Conclusion The Lenti-miR-210-Luciferase is successfully constructed, and miR-210 can promote hPDLSCs the differentiation of angiogenesis.
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Objective: To evaluate the clinical value of the plasma microRNA-155 (miR-155), miR-196a, miR-21 and miR-210 in early diagnosis of patients with pancreatic cancer. Methods: The real-time fluorescent quantitative-PCR was performed to detect the levels of miR-155, miR-196a, miR-21 and miR-210 in plasma samples from sixty patients with pancreatic cancer, twenty patients with chronic pancreatitis, and ten healthy volunteers, as well as the levels of four miRNAs in cancer tissue specimens from ten patients with pancreatic cancer. The serum tumor markers carbohydrate antigen 199 (CA199), CA242 and carcino-embryonic antigen (CEA) were simultaneously detected. The relative expression levels of four miRNAs in plasma among these three groups were compared, and the relationship between miRNA levels in plasma and their levels in pancreatic cancer tissues were statistically analyzed. Finally, the diagnostic efficiency of plasma miR-155, miR-196a, miR-21 and miR-210 for pancreatic cancer was evaluated. Results: The relative expression levels of plasma miR-155, miR-196a, miR-21 and miR-210 in pancreatic cancer group were significantly higher than those in the chronic pancreatitis group and the healthy control group (all P 0.05). The area under receiver operating characteristic (AUC-ROC) curve of plasma miR-155, miR-196a, miR-21 and miR-210 in diagnosis of pancreatic cancer indicated that these four miRNAs had diagnostic value independently (all P < 0.01), in which miR-155 had the highest diagnostic efficiency. The binary Logistic regression model showed that combined detection of plasma miR-196a and miR-210 was more effective in diagnosis of stageI pancreatic cancer as compared with CA199. Conclusion: The plasma miR-155, miR-196a, miR-21 and miR-210 levels are effective to distinguish pancreatic cancer from non-pancreatic cancer. The combined detection of miR-196a and miR-210 may become a promising method for early diagnosis of pancreatic cancer.