MicroRNAs （miRNAs） are small, non-coding RNAs that function as post-transcriptional regulators of gene expression. The deregulated expression of miRNAs is associated with a variety of diseases, including breast canc...MicroRNAs （miRNAs） are small, non-coding RNAs that function as post-transcriptional regulators of gene expression. The deregulated expression of miRNAs is associated with a variety of diseases, including breast cancer. In the present study, we found that miR-495 was markedly up-regulated in clinical breast cancer samples by quantitative real time-PCR （qRT-PCR）. Junctional adhesion molecule A （JAM-A） was predicted to be a potential target of miR-495 by bioinformatics analysis and was subsequently verified by luciferase assay and Western blotting. JAM-A was found to be negatively correlated with the migration of breast cancer cells through loss-of-function and gain-offunction assays, and the inhibition of JAM-A by miR- 495 promoted the migration of MCF-7 and MDA-MB-231 cells. Furthermore, overexpression of JAM-A could restore miR-495-induced breast cancer cell migration. Taken together, our findings suggest that miR-4g5 could facilitate breast cancer progression through the repression of JAM-A, making this miRNA a potential therapeutic target.展开更多
目的探讨miR-495在肝癌组织中的表达以及对肝癌MHCC-97H细胞的影响。方法选取2017年1月至2018年1月我院保存的肝癌组织标本56例(肝癌组),同时选取正常肝脏组织标本40例作为对照组,采用逆转录实时荧光定量PCR(qRT-PCR)检测miR-495表达;...目的探讨miR-495在肝癌组织中的表达以及对肝癌MHCC-97H细胞的影响。方法选取2017年1月至2018年1月我院保存的肝癌组织标本56例(肝癌组),同时选取正常肝脏组织标本40例作为对照组,采用逆转录实时荧光定量PCR(qRT-PCR)检测miR-495表达;选取肝癌MHCC-97H细胞,随机分为对照组、空白质粒组和转染组,其中空白质粒组转染空白质粒,转染组转染miR-495抑制剂,用qRT-PCR检测各组细胞miR-495表达,用CCK法检测各组细胞增殖活性,用Transwell细胞迁移实验检测各组细胞迁移能力。多组间数据比较使用方差分析,进一步两两比较采用LDS-t检验,两组比较使用t检验。结果肝癌组miR-495相对表达量为2.043±0.382,高于对照组,两组比较,差异有统计学意义(P<0.05);临床分期Ⅲ~Ⅳ期、有淋巴结转移患者miR-495相对表达量分别为2.265±0.284和2.290±0.355,高于Ⅰ~Ⅱ期、无淋巴结转移患者(P<0.05),差异有统计学意义;转染组miR-495相对表达量为0.653±0.102,低于对照组和空白质粒组(P<0.05),差异有统计学意义;转染组培养24 h和48 h MHCC-97H细胞A值分别为0.404±0.106和0.604±0.136,低于对照组和空白质粒组(P<0.05);转染组MHCC-97H细胞迁移数为(6.10±1.20)个,低于对照组和空白质粒组(P<0.05)。结论miR-495在肝癌组织中呈高表达,与临床病理特征有一定关系;miR-495对肝癌MHCC-97H细胞增殖以及迁移能力有一定影响。展开更多
Background:Hypoxia is a hallmark of cancer and is associated with poor prognosis.However,the molecular mechanism by which hypoxia promotes tumor progression remains unclear.MicroRNAs dysregulation has been shown to pl...Background:Hypoxia is a hallmark of cancer and is associated with poor prognosis.However,the molecular mechanism by which hypoxia promotes tumor progression remains unclear.MicroRNAs dysregulation has been shown to play a critical role in the tumor and tumor microenvironment.Here,we investigated the roles ofmiR-495 and miR-5688 in human non-small cell lung cancer(NSCLC)and their underlying mechanism.Methods:The expression levels of miR-495 and miR-5688 in human NSCLC tissue specimens were measured by quantitative real-time polymerase chain reaction(qRT-PCR).Deferoxamine(DFO)was used to determine whether the regulation of miR-495 and miR-5688 under hypoxia was dependent on hypoxia-inducible factor 1-alpha(HIF-1α).Furthermore,the functions of miR-495 and miR-5688 in tumor progression were evaluated using colony formation,3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2Htetrazolium(MTS),wound healing,transwell assays,and xenograft model.Two algorithms,PicTAR and Targetscan,were used to predict the target gene of these two miRNAs,and dual-luciferase reporter assay was conducted to confirm the target.The unpaired two-tailed t test,Pearson correlation analysis,and Fisher’s exact probability test were performed for statistical analyses.Results:Two miRNAs,miR-495 and miR-5688,were found to participate in NSCLC progression under hypoxia.They were down-regulated in NSCLC tissues compared with normal tissues.We determined that hypoxia led to the down-regulation of miR-495 and miR-5688 in NSCLC cells,which was independent of HIF-1αand cellular metabolic energy.In addition,miR-495 and miR-5688 suppressed cell proliferation,migration,and invasion in vitro.The NSCLC xenograft model showed that miR-495 and miR-5688 inhibited tumor formation in vivo.Interestingly,we found that miR-495 and miR-5688 had the same target,interleukin-11(IL-11).Recombinant human IL-11 counteracted the effects of miR-495 and miR-5688 on NSCLC cells,suggesting that miR-495 and miR-5688 executed their tumor suppressive role 展开更多
文摘目的 探讨miR-495通过靶向 Notch1信号调控高糖诱导的视神经节细胞(RGCs)凋亡的效果与机制。方法 将高糖培养的RGC-5细胞系随机分为三组:空白组、miR-495阴性对照(miR NC)和miR-495组。miR-NC组与miR-495组分别转染miR-495模拟物阴性对照(miR NC)与miR-495 模拟物(mimic),空白组不进行转染。采用qRT-PCR检测miR-495表达,CCK-8法检测细胞增殖指数,流式细胞术检测检测细胞周期,流式细胞术检测细胞凋亡指数,Western Blot检测Notch1蛋白表达。结果 转染后24 h与36 h,细胞增殖指数显著高于空白组与miR-NC组( P <0.05);细胞凋亡指数显著低于空白组与miR-NC组( P <0.05);转染后36 h,miR-495组的G1期比率相对于miR-NC组和空白组显著降低( P <0.05),S期+G2期比率则显著增加( P <0.05)。转染后24 h与36 h,miR-495组的Notch1蛋白相对表达量显著低于miR-NC组和空白组,空白组与miR-NC组比较差异无统计学意义( P >0.05)。结论 miR-495能减轻高糖诱导的RGCs损伤,促进RGCs增殖,抑制Notch1信号通路,从而调节细胞周期,抑制RGCs凋亡。
文摘MicroRNAs （miRNAs） are small, non-coding RNAs that function as post-transcriptional regulators of gene expression. The deregulated expression of miRNAs is associated with a variety of diseases, including breast cancer. In the present study, we found that miR-495 was markedly up-regulated in clinical breast cancer samples by quantitative real time-PCR （qRT-PCR）. Junctional adhesion molecule A （JAM-A） was predicted to be a potential target of miR-495 by bioinformatics analysis and was subsequently verified by luciferase assay and Western blotting. JAM-A was found to be negatively correlated with the migration of breast cancer cells through loss-of-function and gain-offunction assays, and the inhibition of JAM-A by miR- 495 promoted the migration of MCF-7 and MDA-MB-231 cells. Furthermore, overexpression of JAM-A could restore miR-495-induced breast cancer cell migration. Taken together, our findings suggest that miR-4g5 could facilitate breast cancer progression through the repression of JAM-A, making this miRNA a potential therapeutic target.
文摘目的探讨miR-495在肝癌组织中的表达以及对肝癌MHCC-97H细胞的影响。方法选取2017年1月至2018年1月我院保存的肝癌组织标本56例(肝癌组),同时选取正常肝脏组织标本40例作为对照组,采用逆转录实时荧光定量PCR(qRT-PCR)检测miR-495表达;选取肝癌MHCC-97H细胞,随机分为对照组、空白质粒组和转染组,其中空白质粒组转染空白质粒,转染组转染miR-495抑制剂,用qRT-PCR检测各组细胞miR-495表达,用CCK法检测各组细胞增殖活性,用Transwell细胞迁移实验检测各组细胞迁移能力。多组间数据比较使用方差分析,进一步两两比较采用LDS-t检验,两组比较使用t检验。结果肝癌组miR-495相对表达量为2.043±0.382,高于对照组,两组比较,差异有统计学意义(P<0.05);临床分期Ⅲ~Ⅳ期、有淋巴结转移患者miR-495相对表达量分别为2.265±0.284和2.290±0.355,高于Ⅰ~Ⅱ期、无淋巴结转移患者(P<0.05),差异有统计学意义;转染组miR-495相对表达量为0.653±0.102,低于对照组和空白质粒组(P<0.05),差异有统计学意义;转染组培养24 h和48 h MHCC-97H细胞A值分别为0.404±0.106和0.604±0.136,低于对照组和空白质粒组(P<0.05);转染组MHCC-97H细胞迁移数为(6.10±1.20)个,低于对照组和空白质粒组(P<0.05)。结论miR-495在肝癌组织中呈高表达,与临床病理特征有一定关系;miR-495对肝癌MHCC-97H细胞增殖以及迁移能力有一定影响。
基金supported by the National Natural Science Foundation of China(No.81602026)the Natural Science Foundation of Tianjin(No.18JCQNJC81600 and 18JCZDJC32600).
文摘Background:Hypoxia is a hallmark of cancer and is associated with poor prognosis.However,the molecular mechanism by which hypoxia promotes tumor progression remains unclear.MicroRNAs dysregulation has been shown to play a critical role in the tumor and tumor microenvironment.Here,we investigated the roles ofmiR-495 and miR-5688 in human non-small cell lung cancer(NSCLC)and their underlying mechanism.Methods:The expression levels of miR-495 and miR-5688 in human NSCLC tissue specimens were measured by quantitative real-time polymerase chain reaction(qRT-PCR).Deferoxamine(DFO)was used to determine whether the regulation of miR-495 and miR-5688 under hypoxia was dependent on hypoxia-inducible factor 1-alpha(HIF-1α).Furthermore,the functions of miR-495 and miR-5688 in tumor progression were evaluated using colony formation,3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2Htetrazolium(MTS),wound healing,transwell assays,and xenograft model.Two algorithms,PicTAR and Targetscan,were used to predict the target gene of these two miRNAs,and dual-luciferase reporter assay was conducted to confirm the target.The unpaired two-tailed t test,Pearson correlation analysis,and Fisher’s exact probability test were performed for statistical analyses.Results:Two miRNAs,miR-495 and miR-5688,were found to participate in NSCLC progression under hypoxia.They were down-regulated in NSCLC tissues compared with normal tissues.We determined that hypoxia led to the down-regulation of miR-495 and miR-5688 in NSCLC cells,which was independent of HIF-1αand cellular metabolic energy.In addition,miR-495 and miR-5688 suppressed cell proliferation,migration,and invasion in vitro.The NSCLC xenograft model showed that miR-495 and miR-5688 inhibited tumor formation in vivo.Interestingly,we found that miR-495 and miR-5688 had the same target,interleukin-11(IL-11).Recombinant human IL-11 counteracted the effects of miR-495 and miR-5688 on NSCLC cells,suggesting that miR-495 and miR-5688 executed their tumor suppressive role
