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
Hepatocellular carcinoma (HCC) is the leading cause of cancer death worldwide;nevertheless, currenttherapeutic options are limited or ineffective for many patients. Therefore, elucidation of molecular mechanisms inHCC...Hepatocellular carcinoma (HCC) is the leading cause of cancer death worldwide;nevertheless, currenttherapeutic options are limited or ineffective for many patients. Therefore, elucidation of molecular mechanisms inHCC biology could yield important insights for the intervention of novel therapies. Recently, various studies havereported dysregulation of long non-coding RNAs (lncRNAs) in the initiation and progression of HCC, including H19;however, the biological function of H19 in HCC remains unclear. Here, we show that knockdown of H19 disruptedHCC cell growth, impaired the G1-to-S phase transition, and promoted apoptosis, while overexpression of H19yielded the opposite results. Screening for expression of cell cycle-related genes revealed a significant downregulationof CDK6 at both RNA and protein levels upon H19 suppression. Bioinformatic analysis of the H19 sequence and the3′ untranslated region (3′ UTR) of CDK6 transcripts showed several binding sites for microRNA-107 (miR-107), andthe dual luciferase reporter assay confirmed their direct interaction with miR-107. Consistently, blockage of miR-107activity alleviated the growth suppression phenotypes induced by H19 downregulation, suggesting that H19 serves asa molecular sponge for miR-107 to promote CDK6 expression and cell cycle progression. Together, this studydemonstrates a mechanistic function of H19 in driving the proliferation of HCC cells and suggests H19 suppressionas a novel approach for HCC treatment.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.
基金financially supported by Thailand Science Research and Innovation Fund Chulalongkorn University(CU_FRB65_hea(46)_053_30_34)Ratchadapiseksompotch Fund,Faculty of Medicine,Chulalongkorn University(Grant No.RA 66/017)+1 种基金Thailand Research Fund(TRF)Senior Research Scholar(Grant No.RTA6280004)the Center of Excellence in Hepatitis and Liver Cancer,Faculty of Medicine,Chulalongkorn University.
文摘Hepatocellular carcinoma (HCC) is the leading cause of cancer death worldwide;nevertheless, currenttherapeutic options are limited or ineffective for many patients. Therefore, elucidation of molecular mechanisms inHCC biology could yield important insights for the intervention of novel therapies. Recently, various studies havereported dysregulation of long non-coding RNAs (lncRNAs) in the initiation and progression of HCC, including H19;however, the biological function of H19 in HCC remains unclear. Here, we show that knockdown of H19 disruptedHCC cell growth, impaired the G1-to-S phase transition, and promoted apoptosis, while overexpression of H19yielded the opposite results. Screening for expression of cell cycle-related genes revealed a significant downregulationof CDK6 at both RNA and protein levels upon H19 suppression. Bioinformatic analysis of the H19 sequence and the3′ untranslated region (3′ UTR) of CDK6 transcripts showed several binding sites for microRNA-107 (miR-107), andthe dual luciferase reporter assay confirmed their direct interaction with miR-107. Consistently, blockage of miR-107activity alleviated the growth suppression phenotypes induced by H19 downregulation, suggesting that H19 serves asa molecular sponge for miR-107 to promote CDK6 expression and cell cycle progression. Together, this studydemonstrates a mechanistic function of H19 in driving the proliferation of HCC cells and suggests H19 suppressionas a novel approach for HCC treatment.
