YBX1 gene silencing inhibits migratory and invasive potential via CORO1C in breast cancer in vitro

BackgroundY-box binding protein-1 is an evolutionary conserved transcription and translation regulating protein that is overexpressed in various human malignancies, including breast cancer. Despite reports of YB-1 and its association with distant spread of breast cancer, the intrinsic mechanism underlying this observation remains elusive. This study investigates the role of YB-1 in mediating metastasis in highly invasive breast cancer cell lines.MethodsSilencing the YBX1 gene (which encodes the YB-1 protein) by small interfering RNA (siRNA) was performed in MDA-MB-231 and Hs578T breast cancer cell lines, followed by phenotypic assays including cell migration and invasion assays. Gene expression profiling using Affymetrix GeneChip® Human Transcriptome 2.0 array was subsequently carried out in YB-1 silenced MDA-MB-231 cells. Overexpression and silencing of YBX1 were performed to assess the expression of CORO1C, one of the differentially regulated genes from the transcriptomic analysis. A Gaussia luciferase reporter assay was used to determine if CORO1C is a putative YB-1 downstream target. siRNA-mediated silencing of CORO1C and down-regulation of YBX1 in CORO1C overexpressing MDA-MB-231 cells were performed to evaluate cell migration and invasion.ResultsDownregulation of the YB-1 protein inhibited cell migration and invasion in MDA-MB-231 breast cancer cells. Global gene expression profiling in the YBX1 silenced MDA-MB-231 cells identified differential expression of several genes, including CORO1C (which encodes for an actin binding protein, coronin-1C) as a potential downstream target of YB-1. While knockdown of YBX1 gene decreased CORO1C gene expression, the opposite effects were seen in YB-1 overexpressing cells. Subsequent verification using the reporter assay revealed that CORO1C is an indirect downstream target of YB-1. Silencing of CORO1C by siRNA in MDA-MB-231 cells was also observed to reduce cell migration and invasion. Silencing of YBX1 caused a similar reduction in CORO1C expression, concomitant with a significant decrease in migration in Hs578T cells. In coronin-1C overexpressing MDA-MB-231 cells, increased migration and invasion were abrogated by YB-1 knockdown.ConclusionIt would appear that YB-1 could regulate cell invasion and migration via downregulation of its indirect target coronin-1C. The association between YB-1 and coronin-1C offers a novel approach by which metastasis of breast cancer cells could be targeted and abrogated.

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

[2]  J. Bear,et al.  The role of mammalian coronins in development and disease. , 2008, Sub-cellular biochemistry.

[3]  S. Dunn,et al.  Interplay between YB-1 and IL-6 promotes the metastatic phenotype in breast cancer cells , 2015, Oncotarget.

[4]  F. Bertucci,et al.  Gene expression profiling of breast cell lines identifies potential new basal markers , 2006, Oncogene.

[5]  X. Wang,et al.  Coronin 3 promotes gastric cancer metastasis via the up-regulation of MMP-9 and cathepsin K , 2012, Molecular Cancer.

[6]  Thomas D. Schmittgen,et al.  Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. , 2001, Methods.

[7]  N. Harbeck,et al.  Y‐box factor YB‐1 predicts drug resistance and patient outcome in breast cancer independent of clinically relevant tumor biologic factors HER2, uPA and PAI‐1 , 2002, International journal of cancer.

[8]  B. Bay,et al.  Silencing the YB‐1 Gene Inhibits Cell Migration in Gastric Cancer In Vitro , 2013, Anatomical record.

[9]  Robert A. Weinberg,et al.  Tumor Metastasis: Molecular Insights and Evolving Paradigms , 2011, Cell.

[10]  Yan Li,et al.  Coronin-1C is a novel biomarker for hepatocellular carcinoma invasive progression identified by proteomics analysis and clinical validation , 2010, Journal of experimental & clinical cancer research : CR.

[11]  K. Kohno,et al.  Direct Involvement of the Y-box Binding Protein YB-1 in Genotoxic Stress-induced Activation of the Human Multidrug Resistance 1 Gene* , 1998, The Journal of Biological Chemistry.

[12]  J. Peterse,et al.  Breast cancer metastasis: markers and models , 2005, Nature Reviews Cancer.

[13]  P. Tan,et al.  Y-Box-binding protein-1 is a promising predictive marker of radioresistance and chemoradioresistance in nasopharyngeal cancer , 2009, Modern Pathology.

[14]  Konstantin G. Chernov,et al.  Atomic force microscopy reveals binding of mRNA to microtubules mediated by two major mRNP proteins YB‐1 and PABP , 2008, FEBS letters.

[15]  H. Fernando,et al.  WASP and WAVE proteins: vital intrinsic regulators of cell motility and their role in cancer (review). , 2009, International journal of molecular medicine.

[16]  P. Tan,et al.  Y‐box binding protein 1 is correlated with lymph node metastasis in intestinal‐type gastric cancer , 2015, Histopathology.

[17]  S. Chiu,et al.  YB-1 expression promotes epithelial-to-mesenchymal transition in prostate cancer that is inhibited by a small molecule fisetin , 2014, Oncotarget.

[18]  M. Kessels,et al.  Controlling actin cytoskeletal organization and dynamics during neuronal morphogenesis. , 2011, European journal of cell biology.

[19]  A. Pollock,et al.  YB-1 alters MT1-MMP trafficking and stimulates MCF-7 breast tumor invasion and metastasis. , 2010, Biochemical and biophysical research communications.

[20]  Masaaki Ito,et al.  Characterization of the human hair keratin-associated protein 2 (KRTAP2) gene family. , 2012, The Journal of investigative dermatology.

[21]  Samuel Leung,et al.  Redefining prognostic factors for breast cancer: YB-1 is a stronger predictor of relapse and disease-specific survival than estrogen receptor or HER-2 across all tumor subtypes , 2008, Breast Cancer Research.

[22]  J. Tcherkezian,et al.  Current knowledge of the large RhoGAP family of proteins , 2007, Biology of the cell.

[23]  C. L. Wang.  Caldesmon and the regulation of cytoskeletal functions. , 2008, Advances in experimental medicine and biology.

[24]  J. Bear,et al.  Coronins: the return of the crown. , 2006, Trends in cell biology.

[25]  Y. Sasaguri,et al.  Strong YB‐1 expression is associated with liver metastasis progression and predicts shorter disease‐free survival in advanced gastric cancer , 2012, Journal of surgical oncology.

[26]  T. Triche,et al.  Translational activation of snail1 and other developmentally regulated transcription factors by YB-1 promotes an epithelial-mesenchymal transition. , 2009, Cancer cell.

[27]  J. Bear,et al.  Unraveling the enigma: progress towards understanding the coronin family of actin regulators. , 2011, Trends in cell biology.

[28]  T. Pietsch,et al.  Expression of coronin‐3 (coronin‐1C) in diffuse gliomas is related to malignancy , 2007, The Journal of pathology.

[29]  B. Bay,et al.  Surmounting Chemoresistance by Targeting the Y‐Box Binding Protein‐1 , 2012, Anatomical record.

[30]  B. Goode,et al.  Coronin: the double-edged sword of actin dynamics. , 2008, Sub-cellular biochemistry.

[31]  M. Dietel,et al.  YB-1 provokes breast cancer through the induction of chromosomal instability that emerges from mitotic failure and centrosome amplification. , 2005, Cancer research.

[32]  Hwee Tong Tan,et al.  Identification and functional validation of caldesmon as a potential gastric cancer metastasis-associated protein. , 2013, Journal of proteome research.

[33]  N. Seki,et al.  Downregulation of the microRNA-1/133a cluster enhances cancer cell migration and invasion in lung-squamous cell carcinoma via regulation of Coronin1C , 2014, Journal of Human Genetics.

[34]  Liang Cai,et al.  Coronin 1B Coordinates Arp2/3 Complex and Cofilin Activities at the Leading Edge , 2007, Cell.

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

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

[37]  S. Hellman,et al.  Individual characterisation of the metastatic capacity of human breast carcinoma. , 2000, European journal of cancer.

[38]  J. Bergh,et al.  miR‐206 inhibits cell migration through direct targeting of the actin‐binding protein Coronin 1C in triple‐negative breast cancer , 2014, Molecular oncology.