p63 Attenuates Epithelial to Mesenchymal Potential in an Experimental Prostate Cell Model

The transcription factor p63 is central for epithelial homeostasis and development. In our model of epithelial to mesenchymal transition (EMT) in human prostate cells, p63 was one of the most down-regulated transcription factors during EMT. We therefore investigated the role of p63 in EMT. Over-expression of the predominant epithelial isoform ΔNp63α in mesenchymal type cells of the model led to gain of several epithelial characteristics without resulting in a complete mesenchymal to epithelial transition (MET). This was corroborated by a reciprocal effect when p63 was knocked down in epithelial EP156T cells. Global gene expression analyses showed that ΔNp63α induced gene modules involved in both cell-to-cell and cell-to-extracellular-matrix junctions in mesenchymal type cells. Genome-wide analysis of p63 binding sites using ChIP-seq analyses confirmed binding of p63 to regulatory areas of genes associated with cell adhesion in prostate epithelial cells. DH1 and ZEB1 are two elemental factors in the control of EMT. Over-expression and knock-down of these factors, respectively, were not sufficient alone or in combination with ΔNp63α to reverse completely the mesenchymal phenotype. The partial reversion of epithelial to mesenchymal transition might reflect the ability of ΔNp63α, as a key co-ordinator of several epithelial gene expression modules, to reduce epithelial to mesenchymal plasticity (EMP). The utility of ΔNp63α expression and the potential of reduced EMP in order to counteract metastasis warrant further investigation.

[1]  A. Ashworth,et al.  Genome-wide analysis of p63 binding sites identifies AP-2 factors as co-regulators of epidermal differentiation , 2012, Nucleic acids research.

[2]  Dustin E. Schones,et al.  High-Resolution Profiling of Histone Methylations in the Human Genome , 2007, Cell.

[3]  R. Mantovani,et al.  miR-205 regulates basement membrane deposition in human prostate: implications for cancer development , 2012, Cell Death and Differentiation.

[4]  Clifford A. Meyer,et al.  Model-based Analysis of ChIP-Seq (MACS) , 2008, Genome Biology.

[5]  L. Ellisen,et al.  p63 and p73 in human cancer: defining the network , 2007, Oncogene.

[6]  Kakajan Komurov,et al.  Core epithelial-to-mesenchymal transition interactome gene-expression signature is associated with claudin-low and metaplastic breast cancer subtypes , 2010, Proceedings of the National Academy of Sciences.

[7]  R. Huang,et al.  Epithelial-Mesenchymal Transitions in Development and Disease , 2009, Cell.

[8]  Cole Trapnell,et al.  Ultrafast and memory-efficient alignment of short DNA sequences to the human genome , 2009, Genome Biology.

[9]  A. Levine,et al.  Loss of p63 and its microRNA-205 target results in enhanced cell migration and metastasis in prostate cancer , 2012, Proceedings of the National Academy of Sciences.

[10]  V. Rotter,et al.  Epithelial to Mesenchymal Transition of a Primary Prostate Cell Line with Switches of Cell Adhesion Modules but without Malignant Transformation , 2008, PloS one.

[11]  R. Weinberg,et al.  Roles for microRNAs in the regulation of cell adhesion molecules , 2011, Journal of Cell Science.

[12]  V. Rotter,et al.  Reprogramming of cell junction modules during stepwise epithelial to mesenchymal transition and accumulation of malignant features in vitro in a prostate cell model. , 2011, Experimental cell research.

[13]  M. Shen,et al.  A luminal epithelial stem cell that is a cell of origin for prostate cancer , 2009, Nature.

[14]  V. Rotter,et al.  hTERT-immortalized prostate epithelial and stromal-derived cells: an authentic in vitro model for differentiation and carcinogenesis. , 2006, Cancer research.

[15]  Jason S. Carroll,et al.  p63 regulates an adhesion programme and cell survival in epithelial cells , 2006, Nature Cell Biology.

[16]  Brad T. Sherman,et al.  Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources , 2008, Nature Protocols.

[17]  Bas E. Dutilh,et al.  Genome-Wide Profiling of p63 DNA–Binding Sites Identifies an Element that Regulates Gene Expression during Limb Development in the 7q21 SHFM1 Locus , 2010, PLoS genetics.

[18]  T. H. Bø,et al.  Comparison of nucleic acid targets prepared from total RNA or poly(A) RNA for DNA oligonucleotide microarray hybridization. , 2007, Analytical biochemistry.

[19]  Robert A. Weinberg,et al.  Creation of human tumour cells with defined genetic elements , 1999, Nature.

[20]  R. Vessella,et al.  Comprehensive mutational analysis and mRNA isoform quantification of TP63 in normal and neoplastic human prostate cells , 2009, The Prostate.

[21]  Brad T. Sherman,et al.  Bioinformatics enrichment tools: paths toward the comprehensive functional analysis of large gene lists , 2008, Nucleic acids research.

[22]  Jiaoti Huang,et al.  Identification of a Cell of Origin for Human Prostate Cancer , 2010, Science.

[23]  Jørgen Kjems,et al.  Coordinated epigenetic repression of the miR‐200 family and miR‐205 in invasive bladder cancer , 2011, International journal of cancer.

[24]  S. Ramaswamy,et al.  A Molecular Roadmap of Reprogramming Somatic Cells into iPS Cells , 2012, Cell.

[25]  M. Loda,et al.  p63 regulates commitment to the prostate cell lineage. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[26]  Antoine M. van Oijen,et al.  Real-time single-molecule observation of rolling-circle DNA replication , 2009, Nucleic acids research.

[27]  G. Muto,et al.  BTG2 loss and miR-21 upregulation contribute to prostate cell transformation by inducing luminal markers expression and epithelial–mesenchymal transition , 2013, Oncogene.

[28]  Sandy L. Klemm,et al.  Single-Cell Expression Analyses during Cellular Reprogramming Reveal an Early Stochastic and a Late Hierarchic Phase , 2012, Cell.

[29]  G. Berx,et al.  Regulatory networks defining EMT during cancer initiation and progression , 2013, Nature Reviews Cancer.

[30]  Yi Qu,et al.  Global profiling of histone and DNA methylation reveals epigenetic-based regulation of gene expression during epithelial to mesenchymal transition in prostate cells , 2010, BMC Genomics.

[31]  Graziano Pesole,et al.  Identification and functional characterization of two new transcriptional variants of the human p63 gene , 2009, Nucleic acids research.

[32]  Inge Jonassen,et al.  J-Express: exploring gene expression data using Java , 2001, Bioinform..

[33]  S. Signoretti,et al.  p63 in prostate biology and pathology , 2008, Journal of cellular biochemistry.

[34]  Gareth Browne,et al.  ZEB/miR‐200 feedback loop: At the crossroads of signal transduction in cancer , 2013, International journal of cancer.

[35]  R. Mattingly,et al.  Restoration of E-cadherin cell-cell junctions requires both expression of E-cadherin and suppression of ERK MAP kinase activation in Ras-transformed breast epithelial cells. , 2008, Neoplasia.

[36]  P. Hainaut,et al.  Intraepithelial p63‐dependent expression of distinct components of cell adhesion complexes in normal esophageal mucosa and squamous cell carcinoma , 2010, International journal of cancer.

[37]  Alex E. Lash,et al.  Gene Expression Omnibus: NCBI gene expression and hybridization array data repository , 2002, Nucleic Acids Res..

[38]  Simon S McDade,et al.  Role of ΔNp63γ in Epithelial to Mesenchymal Transition* , 2010, The Journal of Biological Chemistry.

[39]  Yi Qu,et al.  Genome-Wide Profiling of Histone H3 Lysine 4 and Lysine 27 Trimethylation Reveals an Epigenetic Signature in Prostate Carcinogenesis , 2009, PloS one.

[40]  Raja Jothi,et al.  Genome-wide identification of in vivo protein–DNA binding sites from ChIP-Seq data , 2008, Nucleic acids research.