Hepatocellular carcinoma(HCC)remains the third leading cause of cancer-related deaths worldwide;however,its therapeutic options are limited.Understanding the molecular mechanisms of HCC could provide insight into new ...Hepatocellular carcinoma(HCC)remains the third leading cause of cancer-related deaths worldwide;however,its therapeutic options are limited.Understanding the molecular mechanisms of HCC could provide insight into new therapies.Emerging studies indicate the important role of long-noncoding RNAs(lncRNAs)in the pathogenesis of HCC.The expression of the well-studied lncRNA taurine upregulated gene 1(TUG1)is upregulated in HCC tissues,but its transcriptomic effects in HCC cells remain unexplored.We established TUG1-knockdown and control HCC cells for RNA-seq experiments.KEGG analysis revealed glycolysis as the top enriched pathway upon TUG1 silencing.Accordingly,TUG1-depleted HCC cells showed impairments in glucose uptake,ATP synthesis,and lactate production.Clinical HCC tissue data revealed positive gene expression correlations between TUG1 and several glycolysis-related genes.To identify a molecular function of TUG1 in glycolysis,we explored the competing endogenous model and used bioinformatic tools to find the five microRNAs(miRNAs)that had the most binding sites for TUG1.Among these miRNAs,miR-122-5p exhibited an inverse correlation in gene expression with most TUG1-regulated glycolysis genes,including PKM,ALDOA,ENO2,and PFKM.Dual-luciferase assays demonstrated the direct interaction between TUG1 and miR-122-5p and between miR-122-5p and the 3ʹuntranslated regions of both PKM and ALDOA.We further showed that inhibition of miR-122-5p alleviated the suppression of glycolysis induced by TUG1 depletion.Together,our RNA-seq analysis of TUG1-depleted HCC cells,combined with clinical data,reveals a critical role of TUG1 in regulating glycolysis and provides new insight into its oncogenic function in HCC.展开更多
目的探讨检测长链非编码RNA TUG1(long non coding RNA TUG1,Lnc RNA TUG1,TUG1)在冠状动脉粥样硬化性心脏病(冠心病)患者血清中的表达及其临床意义。方法采用实时荧光定量聚合酶链反应(quantitative real time polymemse chain reactio...目的探讨检测长链非编码RNA TUG1(long non coding RNA TUG1,Lnc RNA TUG1,TUG1)在冠状动脉粥样硬化性心脏病(冠心病)患者血清中的表达及其临床意义。方法采用实时荧光定量聚合酶链反应(quantitative real time polymemse chain reaction,QRT-PCR)检测眉山市人民医院收治的106例冠心病阳性患者和98例冠心病阴性对照组的血清TUG1浓度。采用酶联免疫吸附法检测外周血清中胰岛素样生长因子2B(insulin-like growth factor 2B,IGF2B)、白细胞介素-6(interleukin-6,IL-6)、肿瘤坏死因子-α(tumor necrosisfactor-α,TNF-α)的浓度。分析血清TUG1浓度与血清IGF2B、IL-6、TNF-α浓度的相关性。结果与冠心病阴性组比较,冠心病阳性组血清TUG1浓度维持在相对较低的水平;而血清IGF2B、IL-6、TNF-α浓度在冠心病阳性组明显高于冠心病阴性组,差异有统计学意义(P<0.05)。相关分析结果显示,血清TUG1浓度与IGF2B、IL-6及TNF-α浓度呈负相关关系(IGF2B:r=-0.767,P<0.001;IL-6:r=-0.671,P<0.001;TNF-α:r=-0.872,P<0.001)。结论冠心病患者血清TUG1浓度显著降低,可能是潜在的诊断和预测冠心病预后的血清标志物。展开更多
基金supported by the Thailand Science Research and Innovation Fund Chulalongkorn University(Grant No.HEAF67300078)the 90th Anniversary Scholarship,Chulalongkorn University Ratchadapisek Sompoch Fund(Grant No.Batch#55,T.Boonto)the Center of Excellence in Hepatitis and Liver Cancer,Faculty of Medicine,Chulalongkorn University.T.Boonto was supported by the scholarship from the Graduate School,Chulalongkorn University,to commemorate the 72^(nd) anniversary of His Majesty King Bhumibol Adulyadej(Grant No.Batch#22).
文摘Hepatocellular carcinoma(HCC)remains the third leading cause of cancer-related deaths worldwide;however,its therapeutic options are limited.Understanding the molecular mechanisms of HCC could provide insight into new therapies.Emerging studies indicate the important role of long-noncoding RNAs(lncRNAs)in the pathogenesis of HCC.The expression of the well-studied lncRNA taurine upregulated gene 1(TUG1)is upregulated in HCC tissues,but its transcriptomic effects in HCC cells remain unexplored.We established TUG1-knockdown and control HCC cells for RNA-seq experiments.KEGG analysis revealed glycolysis as the top enriched pathway upon TUG1 silencing.Accordingly,TUG1-depleted HCC cells showed impairments in glucose uptake,ATP synthesis,and lactate production.Clinical HCC tissue data revealed positive gene expression correlations between TUG1 and several glycolysis-related genes.To identify a molecular function of TUG1 in glycolysis,we explored the competing endogenous model and used bioinformatic tools to find the five microRNAs(miRNAs)that had the most binding sites for TUG1.Among these miRNAs,miR-122-5p exhibited an inverse correlation in gene expression with most TUG1-regulated glycolysis genes,including PKM,ALDOA,ENO2,and PFKM.Dual-luciferase assays demonstrated the direct interaction between TUG1 and miR-122-5p and between miR-122-5p and the 3ʹuntranslated regions of both PKM and ALDOA.We further showed that inhibition of miR-122-5p alleviated the suppression of glycolysis induced by TUG1 depletion.Together,our RNA-seq analysis of TUG1-depleted HCC cells,combined with clinical data,reveals a critical role of TUG1 in regulating glycolysis and provides new insight into its oncogenic function in HCC.
文摘目的探讨长链非编码RNA(long non-coding RNA,LncRNA)牛磺酸上调基因1(taurine up-regulated gene 1,TUG1)对非小细胞肺癌(non-small cell lung cancer,NSCLC)细胞生物学行为的影响及分子机制。方法采用qRT-PCR、Western blot分别检测组织(NSCLC癌组织、癌旁组织)和细胞(人正常气管上皮细胞HBE及NSCLC细胞A549、H2009、H1975)中TUG1、miR-142-5p及PD-L1的表达。将pcDNA-TUG1、si-TUG1、miR-142-5p mimics、si-TUG1+anti-miR-142-5p分别转染于A549,qRT-PCR、Western blot分别检测细胞中TUG1、miR-142-5p及PD-L1表达;CCK-8检测细胞增殖;Transwell检测细胞迁移、侵袭;双荧光素酶基因实验分别检测TUG1和miR-142-5p、miR-142-5p和PD-L1的关系;RNA免疫共沉淀检测miR-142-5p与TUG1的相互作用。结果NSCLC组织中TUG1(2.28±0.23 vs 1.00±0)、PD-L1(0.93±0.11 vs 0.25±0.01)表达高于癌旁组织,miR-142-5p(0.31±0.02 vs 1.00±0)表达低于癌旁组织(P<0.05);与人正常气管上皮细胞HBE比较,NSCLC细胞A549中TUG1(3.21±0.27 vs 1.00±0)、PD-L1(1.12±0.12 vs 0.24±0.01)表达水平最高,miR-142-5p(0.23±0.02 vs 1.00±0)表达水平最低(P<0.05);过表达TUG1促进A549细胞增殖、迁移、侵袭;下调TUG1或上调miR-142-5p表达均对A549细胞增殖、迁移、侵袭能力发挥抑制作用;TUG1负调控miR-142-5p,miR-142-5p负调控PD-L1;下调miR-142-5p减弱了沉默TUG1对A549细胞增殖、迁移及侵袭的抑制作用。结论TUG1在NSCLC组织和细胞中高表达,沉默TUG1通过miR-142-5p/PD-L1轴抑制A549细胞的增殖、迁移与侵袭。
