Long noncoding RNA HULC modulates the phosphorylation of YB‐1 through serving as a scaffold of extracellular signal–regulated kinase and YB‐1 to enhance hepatocarcinogenesis

Dysregulated expression of long noncoding RNAs has been reported in many types of cancers, indicating that it may play a critical role in tumorigenesis. The long noncoding RNA highly up‐regulated in liver cancer (HULC) was first characterized in hepatocellular carcinoma. However, the detailed mechanisms of HULC remain unclear. Here, we demonstrate a novel mechanism by which long noncoding RNA plays oncogenic roles through modulating the phosphorylation status of its interaction protein. First, we validated the markedly increased expression levels of HULC in hepatocellular carcinoma tissues compared to their adjacent noncancerous tissues. Furthermore, up‐regulation of HULC was correlated with grading and overall survival. Meanwhile, HULC could promote cell proliferation, migration, and invasion in vitro and inhibit cisplatin‐induced apoptosis. Moreover, we show that HULC specifically binds to Y‐box binding protein 1 (YB‐1) protein both in vitro and in vivo. YB‐1 is a major component of translationally inactive messenger ribonucleoprotein particles which keeps mRNA in a silent state. Our study further demonstrated that HULC could promote the phosphorylation of YB‐1 protein, which leads to the release of YB‐1 from its bound mRNA. As a consequence, translation of silenced oncogenic mRNAs would be activated, including cyclin D1, cyclin E1, and matrix metalloproteinase 3. In addition, we found that HULC promotes the phosphorylation of YB‐1 protein mainly through extracellular signal–regulated kinase. Conclusion: We demonstrate that HULC promotes the phosphorylation of YB‐1 through the extracellular signal–regulated kinase pathway, in turn regulates the interaction of YB‐1 with certain oncogenic mRNAs, and consequently accelerates the translation of these mRNAs in the process of tumorigenesis. (Hepatology 2017;65:1612‐1627)

[1]  M. Rubin,et al.  Detection of TMPRSS2-ERG fusion gene expression in prostate cancer specimens by a novel assay using branched DNA. , 2009, Urology.

[2]  D. Hanahan,et al.  Hallmarks of Cancer: The Next Generation , 2011, Cell.

[3]  Meng-Han Lin,et al.  The development of a method of suspension RNA-FISH for forensically relevant epithelial cells using LNA probes. , 2014, Forensic science international. Genetics.

[4]  Howard Y. Chang,et al.  Long noncoding RNA HOTAIR reprograms chromatin state to promote cancer metastasis , 2010, Nature.

[5]  C. Print,et al.  YB-1: oncoprotein, prognostic marker and therapeutic target? , 2013, The Biochemical journal.

[6]  L. Ovchinnikov,et al.  YB‐1 protein: functions and regulation , 2014, Wiley interdisciplinary reviews. RNA.

[7]  Sonia Sharma,et al.  Dephosphorylation of the nuclear factor of activated T cells (NFAT) transcription factor is regulated by an RNA-protein scaffold complex , 2011, Proceedings of the National Academy of Sciences.

[8]  H. Takano,et al.  Enhanced coexpression of YB-1 and DNA topoisomerase II alpha genes in human colorectal carcinomas. , 1999, International journal of cancer.

[9]  Yong-Yeon Cho,et al.  Y-box binding protein-1 serine 102 is a downstream target of p90 ribosomal S6 kinase in basal-like breast cancer cells , 2008, Breast Cancer Research.

[10]  Xiao Xu,et al.  The xCELLigence system for real-time and label-free monitoring of cell viability. , 2011, Methods in molecular biology.

[11]  F. Costa,et al.  Non-coding RNAs: new players in eukaryotic biology. , 2005, Gene.

[12]  J. Massagué,et al.  G1 cell-cycle control and cancer , 2004, Nature.

[13]  C. Mathers,et al.  Cancer incidence and mortality worldwide: Sources, methods and major patterns in GLOBOCAN 2012 , 2015, International journal of cancer.

[14]  Li-bin Yang,et al.  Increased expression of lncRNA HULC indicates a poor prognosis and promotes cell metastasis in osteosarcoma. , 2015, International journal of clinical and experimental pathology.

[15]  Xuehao Wang,et al.  LncRNA HULC affects the differentiation of Treg in HBV-related liver cirrhosis. , 2015, International immunopharmacology.

[16]  P. Cohen,et al.  Identification of Regulatory Phosphorylation Sites in Mitogen-activated Protein Kinase (MAPK)-activated Protein Kinase-1a/p90 rsk That Are Inducible by MAPK* , 1998, The Journal of Biological Chemistry.

[17]  G. Goodall,et al.  Phosphorylation of cold shock domain/Y‐box proteins by ERK2 and GSK3β and repression of the human VEGF promoter , 2005, FEBS letters.

[18]  K. Zatloukal,et al.  Characterization of HULC, a novel gene with striking up-regulation in hepatocellular carcinoma, as noncoding RNA. , 2007, Gastroenterology.

[19]  L. Ye,et al.  Long non-coding RNA HULC promotes tumor angiogenesis in liver cancer by up-regulating sphingosine kinase 1 (SPHK1) , 2015, Oncotarget.

[20]  Sevim Ozgur,et al.  Posttranscriptional destabilization of the liver‐specific long noncoding RNA HULC by the IGF2 mRNA‐binding protein 1 (IGF2BP1) , 2013, Hepatology.

[21]  J. Mullins,et al.  Production of large numbers of mitotic mammalian cells by use of the reversible microtubule inhibitor nocodazole. Nocodazole accumulated mitotic cells. , 1980, Experimental cell research.

[22]  S. Arii,et al.  The Up-Regulation of Y-Box Binding Proteins (DNA Binding Protein A and Y-Box Binding Protein-1) as Prognostic Markers of Hepatocellular Carcinoma , 2005, Clinical Cancer Research.

[23]  Hongwei Ma,et al.  Plasma HULC as a Promising Novel Biomarker for the Detection of Hepatocellular Carcinoma , 2013, BioMed research international.

[24]  H. Yao,et al.  A cytoplasmic NF-κB interacting long noncoding RNA blocks IκB phosphorylation and suppresses breast cancer metastasis. , 2015, Cancer cell.

[25]  Ronald A. DePinho,et al.  Hepatocellular carcinoma pathogenesis: from genes to environment , 2006, Nature Reviews Cancer.

[26]  G. Chrousos,et al.  Noncoding RNA Gas5 Is a Growth Arrest– and Starvation-Associated Repressor of the Glucocorticoid Receptor , 2010, Science Signaling.

[27]  P. Kaldis,et al.  Cdks, cyclins and CKIs: roles beyond cell cycle regulation , 2013, Development.

[28]  G. Ifere,et al.  Prostate cancer gene expression marker 1 (PCGEM1): a patented prostate- specific non-coding gene and regulator of prostate cancer progression. , 2009, Recent patents on DNA & gene sequences.

[29]  Xuetao Cao,et al.  The STAT3-Binding Long Noncoding RNA lnc-DC Controls Human Dendritic Cell Differentiation , 2014, Science.

[30]  H. Takano,et al.  Enhanced coexpression of YB‐1 and DNA topoisomerase II α genes in human colorectal carcinomas , 1999 .

[31]  M. Heikenwalder,et al.  The direct and indirect roles of HBV in liver cancer: prospective markers for HCC screening and potential therapeutic targets , 2015, The Journal of pathology.

[32]  Jiayi Wang,et al.  CREB up-regulates long non-coding RNA, HULC expression through interaction with microRNA-372 in liver cancer , 2010, Nucleic acids research.

[33]  Abraham Hochberg,et al.  Highly upregulated in liver cancer noncoding RNA is overexpressed in hepatic colorectal metastasis , 2009, European journal of gastroenterology & hepatology.

[34]  J. Rinn,et al.  A Large Intergenic Noncoding RNA Induced by p53 Mediates Global Gene Repression in the p53 Response , 2010, Cell.

[35]  A. Wolffe.  Structural and functional properties of the evolutionarily ancient Y‐box family of nucleic acid binding proteins , 1994, BioEssays : news and reviews in molecular, cellular and developmental biology.

[36]  L. Ovchinnikov,et al.  Y-box-binding protein 1 (YB-1) and its functions , 2011, Biochemistry (Moscow).

[37]  N. Sonenberg,et al.  Akt-Mediated YB-1 Phosphorylation Activates Translation of Silent mRNA Species , 2006, Molecular and Cellular Biology.

[38]  B. Blencowe,et al.  The nuclear-retained noncoding RNA MALAT1 regulates alternative splicing by modulating SR splicing factor phosphorylation. , 2010, Molecular cell.

[39]  M. Gleave,et al.  YB‐1 is upregulated during prostate cancer tumor progression and increases P‐glycoprotein activity , 2004, The Prostate.

[40]  Wei Gao,et al.  Long noncoding RNA HULC is a novel biomarker of poor prognosis in patients with pancreatic cancer , 2014, Medical Oncology.

[41]  Andrew D. Hamilton,et al.  Akt/Protein Kinase B Signaling Inhibitor-2, a Selective Small Molecule Inhibitor of Akt Signaling with Antitumor Activity in Cancer Cells Overexpressing Akt , 2004, Cancer Research.

[42]  B. Sarcevic,et al.  Control of cell cycle progression by phosphorylation of cyclin-dependent kinase (CDK) substrates. , 2010, Bioscience reports.

[43]  E. Nishida,et al.  ERK MAP kinase in G1 cell cycle progression and cancer , 2006, Cancer science.

[44]  R. Eils,et al.  Etiology‐dependent molecular mechanisms in human hepatocarcinogenesis , 2007, Hepatology.

[45]  M. Gleave,et al.  Molecular Decoy to the Y-Box Binding Protein-1 Suppresses the Growth of Breast and Prostate Cancer Cells whilst Sparing Normal Cell Viability , 2010, PloS one.

[46]  J. Rinn,et al.  Modular regulatory principles of large non-coding RNAs , 2012, Nature.

[47]  E. Yorida,et al.  Akt phosphorylates the Y-box binding protein 1 at Ser102 located in the cold shock domain and affects the anchorage-independent growth of breast cancer cells , 2005, Oncogene.

[48]  Yumei Du,et al.  Elevation of Highly Up-regulated in Liver Cancer (HULC) by Hepatitis B Virus X Protein Promotes Hepatoma Cell Proliferation via Down-regulating p18* , 2012, The Journal of Biological Chemistry.

[49]  C. Kanduri.  Long noncoding RNA and epigenomics. , 2011, Advances in experimental medicine and biology.

[50]  Howard Y. Chang,et al.  Long Noncoding RNA as Modular Scaffold of Histone Modification Complexes , 2010, Science.

[51]  T. Ishikawa.  Future perspectives on the treatment of hepatocellular carcinoma with cisplatin. , 2009, World journal of hepatology.