MicroRNAs （miRNAs or miRs） are a class of short, non-coding RNAs that participate in various oncological processes. This study aims to explore the roles of microRNA-34a （miR-34a） in invasive urothelial bladder car...MicroRNAs （miRNAs or miRs） are a class of short, non-coding RNAs that participate in various oncological processes. This study aims to explore the roles of microRNA-34a （miR-34a） in invasive urothelial bladder carcinoma. miR-34a was transfected into bladder cancer cell lines 253J and J82. The miR-34a expression levels in tissues and cells were detected by using qRT-PCR. The Notch1 expression was detected by qRT-PCR and Western blotting. Cell migratory and invasive abilities were measured by Transwell chamber assay. Bioinformatics and luciferase assay were performed to predict and analyze the binding sites between miRNA-34a and Notch1. It was found that there was aberrant expression of miR-34a in bladder cancer tissues. Moreover, we revealed that ectopic expression of miR-34a suppressed cell migration and invasion, while forced expression of Notch1 increased cell migratory and invasive abilities. Finally, we observed that miR-34a transfection significantly down-regulated luciferase activity and reduced the mRNA and protein levels of Notch1. Our study concluded that microRNA-34a antagonizes Notch1 and inhibits cell migration and invasion of bladder cancer cells, which indicates the tumor-suppressive function of microRNA-34a in bladder cancer.展开更多
AIM： To investigate the role of micro RNA-34a（mi R-34a） in the induction of apoptosis of human lens epithelial（HLE-B3） cells. METHODS： The apoptosis of HLE-B3 cells was detected by Annexin V-PE apoptosis detecti...AIM： To investigate the role of micro RNA-34a（mi R-34a） in the induction of apoptosis of human lens epithelial（HLE-B3） cells. METHODS： The apoptosis of HLE-B3 cells was detected by Annexin V-PE apoptosis detection kit after the treatment with 200 μmol/L H2O2 for 24h and lentiviral mi R-34 a vector transfection. The expression of mi R-34 a in the cells was quantified by quantitative real time polymerase chain reaction（q RT-PCR） in response to H2O2 exposure and the vector transfection. The effects of overexpression of mi R-34 a on the expression of B-cell lymphoma-2（Bcl-2） and silent information regulator 1（SIRT1） was determined by q RT-PCR and Western blot. RESULTS： The expression of mi R-34 a was up-regulated by the treatment of H2O2 in HLE-B3 cells. The increased expression of mi R-34 a is accompanied with the cell apoptosis. Consistence with the H2O2 exposure,ectopic overexpression of mi R-34 a in HLE-B3 cells promoted cells apoptosis. Importantly the anti-apoptosis factors Bcl-2 and SIRT1 were reduced significantly by up-regulation of mi R-34 a in HLE-B3 cells.CONCLUSION： Mi R-34 a promotes the apoptosis of HLE-B3 cells by down-regulating Bcl-2 and SIRT1,suggesting that mi R-34 a may involve in the pathogenesis of cataract formation and targeting mi R-34 a may be a potentially therapeutic approach for treatment of cataract.展开更多
BACKGROUND: Notch-1/NF-κB signaling plays a key role in the cecal ligation and puncture(CLP)-induced sepsis. This study aims to investigate the intervention effects of microRNA-34a(miR-34a) lentivirus regulating Notc...BACKGROUND: Notch-1/NF-κB signaling plays a key role in the cecal ligation and puncture(CLP)-induced sepsis. This study aims to investigate the intervention effects of microRNA-34a(miR-34a) lentivirus regulating Notch-1/NF-κB signaling pathway on lipopolysaccharide(LPS)-induced human umbilical vein endothelial cells(HUVEC).METHODS: HUVEC were divided into four groups as the following: they were infected with negative control lentivirus(NC group) or miR-34a lentivirus(OE group); LPS(1 g/mL) was added on the third day on the basis of NC group and OE group for 24 hours(NC+LPS group or OE+LPS group). The levels of TNF-α, IL-1β, IL-6, and IL-10 in the cell supernatants, and the mRNA and protein expression of Notch-1 and NF-κB in the HUVEC were evaluated.RESULTS: After 24 hours, the levels of TNF-α, IL-1β, IL-6 in the cell supernatants and the protein expression of NF-κB from NC+LPS group were significantly higher than those of NC group, but IL-10 level and the protein expression of Notch-1 in NC+LPS group were the opposite. After intervention of miR-34a lentivirus, the cell supernatants TNF-α and the protein expression of NF-κB in OE+LPS group after 24 hours markedly decreased compared to NC+LPS group. While the cell supernatants IL-1β and IL-6 and the mRNA expression of NF-κB slightly decreased in OE+LPS group, IL-10 and the mRNA and protein expression of Notch-1 were the opposite.CONCLUSION: miR-34a regulating Notch-1/NF-κB signaling pathway can reduce the HUVEC damage caused by LPS stimulation.展开更多
Objective: To investigate the effect and underlying mechanisms of Tiaoxin Recipe(a Chinese herbal formula) treatment on Alzheimer's disease(AD).Methods: Twelve-week-old APPswe/PS1 DE9(APP/PS1) double transgenic mi...Objective: To investigate the effect and underlying mechanisms of Tiaoxin Recipe(a Chinese herbal formula) treatment on Alzheimer's disease(AD).Methods: Twelve-week-old APPswe/PS1 DE9(APP/PS1) double transgenic mice were used as a model of AD-afflicted mice. One group of mice was treated with Tiaoxin Recipe by gastrogavage for 12 weeks,while two other groups were given intraperitoneal injections of nicotinamide adenine dinucleotide or FK866 for 4 weeks. Morris water maze and thioflavin S staining tests were performed to evaluate cognitive impairment and amyloid plaque deposition, respectively. Serum amyloid-β1-42(Aβ1-42) content was detected using an enzyme-linked immunosorbent assay, and quantitative reverse transcriptionpolymerase chain reaction was performed to examine the expression levels of microRNA-34 a(miR-34 a) in cortex and hippocampus samples of the study mice.Results: Compared with the normal control group, the memory and learning abilities of the APP/PS1 model group were found to be impaired(P < 0.01), as shown by the increased levels of senile plaque deposition in cortex and hippocampus(P < 0.01), miR-34 a expression(P < 0.01) and serum Aβ1-42 content(P < 0.01). Treatment with Tiaoxin Recipe significantly reduced memory impairment(P < 0.01) by reducing amyloid plaque accumulation in cortex and hippocampus(P < 0.01), miR-34 a expression(P < 0.01) and serum Aβ1-42 content(P < 0.01) in APP/PS1 mice.Conclusion: Tiaoxin Recipe is a viable complementary or alternative therapeutic treatment that is capable of delaying the development of early-stage AD by inhibiting the expression of miR-34 a.展开更多
The effects of microRNA-34a （miR-34a）-regulated Notch1 gene on the proliferation and apoptosis of the human glioma cell line U87 were investigated in this study. The U87 cells were divided into miR-34a mimics, negat...The effects of microRNA-34a （miR-34a）-regulated Notch1 gene on the proliferation and apoptosis of the human glioma cell line U87 were investigated in this study. The U87 cells were divided into miR-34a mimics, negative control, mock transfection and blank control groups in terms of different treatments. In miR-34a mimics group, human U87 glioma cells were transfected with miR-34a mimics by using lipofectamine 2000. The cells transfected with nonsense microRNA were set up as negative control group. Those treated with lipofectamine 2000 only were designated to the mock tranfection group. In the blank control group, the cells were cultured routinely and no treatment was given. The expression of miR-34a and Notch1 was detected by using real-time RT-PCR. Western blotting was employed to monitor the change in Notch1 protein. Cell proliferation and apoptosis were measured by CCK-8 and flow cytometry. The results showed that the proliferative ability of U87 cells was significantly reduced and the apoptotic cells increased in miR-34a mimics group relative to control groups. The expression of miR-34a was significantly up-regulated in mimics group as compared with control groups （P〈0.05）. Furthermore, Notch1 protein levels were significantly decreased in miR-34a mimics group when compared with control groups （P〈0.05）, but the mRNA expression of Notch1 showed no significant difference among these groups. It was concluded that miR-34a may suppress the proliferation and induce apoptosis of U87 cells by decreasing the expression of target gene Notch1, suggesting that miR-34a may become a promising gene therapeutic target for brain glioma.展开更多
Granulosa cells（GCs） are somatic cells of ovary, the behaviors of GCs are important for ovarian function. MicroRNAs（miRNAs） are a class of endogenous 18–24 nucleotide（nt） non-coding RNAs, some of which have bee...Granulosa cells（GCs） are somatic cells of ovary, the behaviors of GCs are important for ovarian function. MicroRNAs（miRNAs） are a class of endogenous 18–24 nucleotide（nt） non-coding RNAs, some of which have been shown to be important regulators of GCs function. miR-34c involved in the regulation of various biological processes and was identified to be a pro-apoptotic and anti-proliferative factor in many cell types. However, the roles of miR-34c in GCs function remain unknown. In this study, we used Annexin V-FITC and Ed U assays to demonstrate that miR-34c exerted pro-apoptotic and anti-proliferative effects in porcine GCs. Dual-luciferase reporter assays, quantitative real-time PCR（q RT-PCR） and Western blotting identified Forkhead box O3a（Fox O3a） as a direct target gene of miR-34c. The overexpression of FoxO3a rescued the phenotypic change caused by miR-34c in porcine GCs. In conclusion, miR-34c regulate the function of porcine GCs by targeting FoxO3a.展开更多
目的探讨脓毒症休克患者血清微小核糖核酸miR-214、miR-34b和miR-22的表达特征,及其与患者病情程度及预后的关系。方法选取2020年1月至2024年1月南阳市中心医院感染科收治的脓毒症休克患者128例作为研究对象,根据预后分为存活组(n=67)...目的探讨脓毒症休克患者血清微小核糖核酸miR-214、miR-34b和miR-22的表达特征,及其与患者病情程度及预后的关系。方法选取2020年1月至2024年1月南阳市中心医院感染科收治的脓毒症休克患者128例作为研究对象,根据预后分为存活组(n=67)和死亡组(n=61),另选同期收治的未休克脓毒症患者130例作为对照,采用qRT-PCR法检测各组血清miR-214、miR-34b和miR-22表达水平。通过pearson相关分析评估各组血清miRNAs水平与急性生理学与慢性健康状况(Acute Physiology and Chronic Health Status Evaluation,APACHEⅡ)评分和序贯器官衰竭(Sequential Organ Failure,SOFA)评分的相关性,采用多因素Logistic回归分析预后影响因素,并绘制受试者工作特征(ROC)曲线评估血清mi RNAs对患者预后的预测效能。结果与无休克组相比,休克组血清miR-214、miR-22表达水平降低,miR-34b表达水平升高,差异具有统计学意义(P<0.05);miR-214、miR-22与APACHEⅡ评分和SOFA评分呈负相关,miR-34b与APACHEⅡ评分和SOFA评分呈正相关(P<0.05);多因素分析显示SOFA评分、休克指数及血清miR-34b均为独立危险因素(P<0.05),miR-214和miR-22是保护因素(P<0.05);ROC曲线评估结果显示miR-214、miR-34b、miR-22以及联合预测患者死亡的AUC分别为0.838、0.812、0.825和0.922,联合预测优于单独预测。结论脓毒症休克患者血清miR-214和miR-22表达下调,miR-34b表达上调,其表达水平与患者疾病严重程度密切相关。三种miRNAs联合检测对脓毒症休克患者预后具有较高的预测价值,可为临床病情评估和预后判断提供新的分子标志物。展开更多
基金supported by a grant from the National Natural Science Foundation of China(No.30972982)
文摘MicroRNAs （miRNAs or miRs） are a class of short, non-coding RNAs that participate in various oncological processes. This study aims to explore the roles of microRNA-34a （miR-34a） in invasive urothelial bladder carcinoma. miR-34a was transfected into bladder cancer cell lines 253J and J82. The miR-34a expression levels in tissues and cells were detected by using qRT-PCR. The Notch1 expression was detected by qRT-PCR and Western blotting. Cell migratory and invasive abilities were measured by Transwell chamber assay. Bioinformatics and luciferase assay were performed to predict and analyze the binding sites between miRNA-34a and Notch1. It was found that there was aberrant expression of miR-34a in bladder cancer tissues. Moreover, we revealed that ectopic expression of miR-34a suppressed cell migration and invasion, while forced expression of Notch1 increased cell migratory and invasive abilities. Finally, we observed that miR-34a transfection significantly down-regulated luciferase activity and reduced the mRNA and protein levels of Notch1. Our study concluded that microRNA-34a antagonizes Notch1 and inhibits cell migration and invasion of bladder cancer cells, which indicates the tumor-suppressive function of microRNA-34a in bladder cancer.
文摘AIM： To investigate the role of micro RNA-34a（mi R-34a） in the induction of apoptosis of human lens epithelial（HLE-B3） cells. METHODS： The apoptosis of HLE-B3 cells was detected by Annexin V-PE apoptosis detection kit after the treatment with 200 μmol/L H2O2 for 24h and lentiviral mi R-34 a vector transfection. The expression of mi R-34 a in the cells was quantified by quantitative real time polymerase chain reaction（q RT-PCR） in response to H2O2 exposure and the vector transfection. The effects of overexpression of mi R-34 a on the expression of B-cell lymphoma-2（Bcl-2） and silent information regulator 1（SIRT1） was determined by q RT-PCR and Western blot. RESULTS： The expression of mi R-34 a was up-regulated by the treatment of H2O2 in HLE-B3 cells. The increased expression of mi R-34 a is accompanied with the cell apoptosis. Consistence with the H2O2 exposure,ectopic overexpression of mi R-34 a in HLE-B3 cells promoted cells apoptosis. Importantly the anti-apoptosis factors Bcl-2 and SIRT1 were reduced significantly by up-regulation of mi R-34 a in HLE-B3 cells.CONCLUSION： Mi R-34 a promotes the apoptosis of HLE-B3 cells by down-regulating Bcl-2 and SIRT1,suggesting that mi R-34 a may involve in the pathogenesis of cataract formation and targeting mi R-34 a may be a potentially therapeutic approach for treatment of cataract.
基金supported by a grant from Natural Science Foundation of Zhejiang Province of China(LY14H150003)
文摘BACKGROUND: Notch-1/NF-κB signaling plays a key role in the cecal ligation and puncture(CLP)-induced sepsis. This study aims to investigate the intervention effects of microRNA-34a(miR-34a) lentivirus regulating Notch-1/NF-κB signaling pathway on lipopolysaccharide(LPS)-induced human umbilical vein endothelial cells(HUVEC).METHODS: HUVEC were divided into four groups as the following: they were infected with negative control lentivirus(NC group) or miR-34a lentivirus(OE group); LPS(1 g/mL) was added on the third day on the basis of NC group and OE group for 24 hours(NC+LPS group or OE+LPS group). The levels of TNF-α, IL-1β, IL-6, and IL-10 in the cell supernatants, and the mRNA and protein expression of Notch-1 and NF-κB in the HUVEC were evaluated.RESULTS: After 24 hours, the levels of TNF-α, IL-1β, IL-6 in the cell supernatants and the protein expression of NF-κB from NC+LPS group were significantly higher than those of NC group, but IL-10 level and the protein expression of Notch-1 in NC+LPS group were the opposite. After intervention of miR-34a lentivirus, the cell supernatants TNF-α and the protein expression of NF-κB in OE+LPS group after 24 hours markedly decreased compared to NC+LPS group. While the cell supernatants IL-1β and IL-6 and the mRNA expression of NF-κB slightly decreased in OE+LPS group, IL-10 and the mRNA and protein expression of Notch-1 were the opposite.CONCLUSION: miR-34a regulating Notch-1/NF-κB signaling pathway can reduce the HUVEC damage caused by LPS stimulation.
基金supported by grants from the National Natural Science Foundation of China (No. 81503626)the Shanghai Health Bureau Youth Fund (No. 201540254)
文摘Objective: To investigate the effect and underlying mechanisms of Tiaoxin Recipe(a Chinese herbal formula) treatment on Alzheimer's disease(AD).Methods: Twelve-week-old APPswe/PS1 DE9(APP/PS1) double transgenic mice were used as a model of AD-afflicted mice. One group of mice was treated with Tiaoxin Recipe by gastrogavage for 12 weeks,while two other groups were given intraperitoneal injections of nicotinamide adenine dinucleotide or FK866 for 4 weeks. Morris water maze and thioflavin S staining tests were performed to evaluate cognitive impairment and amyloid plaque deposition, respectively. Serum amyloid-β1-42(Aβ1-42) content was detected using an enzyme-linked immunosorbent assay, and quantitative reverse transcriptionpolymerase chain reaction was performed to examine the expression levels of microRNA-34 a(miR-34 a) in cortex and hippocampus samples of the study mice.Results: Compared with the normal control group, the memory and learning abilities of the APP/PS1 model group were found to be impaired(P < 0.01), as shown by the increased levels of senile plaque deposition in cortex and hippocampus(P < 0.01), miR-34 a expression(P < 0.01) and serum Aβ1-42 content(P < 0.01). Treatment with Tiaoxin Recipe significantly reduced memory impairment(P < 0.01) by reducing amyloid plaque accumulation in cortex and hippocampus(P < 0.01), miR-34 a expression(P < 0.01) and serum Aβ1-42 content(P < 0.01) in APP/PS1 mice.Conclusion: Tiaoxin Recipe is a viable complementary or alternative therapeutic treatment that is capable of delaying the development of early-stage AD by inhibiting the expression of miR-34 a.
基金supported by grants from the Key Clinic Programs of Ministry of Health(2010-2012)the 11th Five-year Plan of National Science and Technology Supporting Project(No.2006BAI05A07)
文摘The effects of microRNA-34a （miR-34a）-regulated Notch1 gene on the proliferation and apoptosis of the human glioma cell line U87 were investigated in this study. The U87 cells were divided into miR-34a mimics, negative control, mock transfection and blank control groups in terms of different treatments. In miR-34a mimics group, human U87 glioma cells were transfected with miR-34a mimics by using lipofectamine 2000. The cells transfected with nonsense microRNA were set up as negative control group. Those treated with lipofectamine 2000 only were designated to the mock tranfection group. In the blank control group, the cells were cultured routinely and no treatment was given. The expression of miR-34a and Notch1 was detected by using real-time RT-PCR. Western blotting was employed to monitor the change in Notch1 protein. Cell proliferation and apoptosis were measured by CCK-8 and flow cytometry. The results showed that the proliferative ability of U87 cells was significantly reduced and the apoptotic cells increased in miR-34a mimics group relative to control groups. The expression of miR-34a was significantly up-regulated in mimics group as compared with control groups （P〈0.05）. Furthermore, Notch1 protein levels were significantly decreased in miR-34a mimics group when compared with control groups （P〈0.05）, but the mRNA expression of Notch1 showed no significant difference among these groups. It was concluded that miR-34a may suppress the proliferation and induce apoptosis of U87 cells by decreasing the expression of target gene Notch1, suggesting that miR-34a may become a promising gene therapeutic target for brain glioma.
基金supported by the National Natural Science Foundation of China (31201771)the earmarked fund for the China Agriculture Research System (CARS-36)
文摘Granulosa cells（GCs） are somatic cells of ovary, the behaviors of GCs are important for ovarian function. MicroRNAs（miRNAs） are a class of endogenous 18–24 nucleotide（nt） non-coding RNAs, some of which have been shown to be important regulators of GCs function. miR-34c involved in the regulation of various biological processes and was identified to be a pro-apoptotic and anti-proliferative factor in many cell types. However, the roles of miR-34c in GCs function remain unknown. In this study, we used Annexin V-FITC and Ed U assays to demonstrate that miR-34c exerted pro-apoptotic and anti-proliferative effects in porcine GCs. Dual-luciferase reporter assays, quantitative real-time PCR（q RT-PCR） and Western blotting identified Forkhead box O3a（Fox O3a） as a direct target gene of miR-34c. The overexpression of FoxO3a rescued the phenotypic change caused by miR-34c in porcine GCs. In conclusion, miR-34c regulate the function of porcine GCs by targeting FoxO3a.
文摘目的探讨脓毒症休克患者血清微小核糖核酸miR-214、miR-34b和miR-22的表达特征,及其与患者病情程度及预后的关系。方法选取2020年1月至2024年1月南阳市中心医院感染科收治的脓毒症休克患者128例作为研究对象,根据预后分为存活组(n=67)和死亡组(n=61),另选同期收治的未休克脓毒症患者130例作为对照,采用qRT-PCR法检测各组血清miR-214、miR-34b和miR-22表达水平。通过pearson相关分析评估各组血清miRNAs水平与急性生理学与慢性健康状况(Acute Physiology and Chronic Health Status Evaluation,APACHEⅡ)评分和序贯器官衰竭(Sequential Organ Failure,SOFA)评分的相关性,采用多因素Logistic回归分析预后影响因素,并绘制受试者工作特征(ROC)曲线评估血清mi RNAs对患者预后的预测效能。结果与无休克组相比,休克组血清miR-214、miR-22表达水平降低,miR-34b表达水平升高,差异具有统计学意义(P<0.05);miR-214、miR-22与APACHEⅡ评分和SOFA评分呈负相关,miR-34b与APACHEⅡ评分和SOFA评分呈正相关(P<0.05);多因素分析显示SOFA评分、休克指数及血清miR-34b均为独立危险因素(P<0.05),miR-214和miR-22是保护因素(P<0.05);ROC曲线评估结果显示miR-214、miR-34b、miR-22以及联合预测患者死亡的AUC分别为0.838、0.812、0.825和0.922,联合预测优于单独预测。结论脓毒症休克患者血清miR-214和miR-22表达下调,miR-34b表达上调,其表达水平与患者疾病严重程度密切相关。三种miRNAs联合检测对脓毒症休克患者预后具有较高的预测价值,可为临床病情评估和预后判断提供新的分子标志物。
