Quantitative analysis of ERG expression and its splice isoforms in formalin-fixed, paraffin-embedded prostate cancer samples: association with seminal vesicle invasion and biochemical recurrence.

OBJECTIVES The proto-oncogene ETS-related gene (ERG) is consistently overexpressed in prostate cancer. Alternatively spliced isoforms of ERG have variable biological activities; inclusion of exon 11 (72 base pairs [bp]) is associated with aggressiveness and progression of disease. Exon 10 (81 bp) has also been shown to be alternatively spliced. Within this study, we assess whether ERG protein, messenger RNA (mRNA), and ERG splice isoform mRNA expression is altered as prostate cancer progresses. METHODS Detection of the TMPRSS2-ERG fusion was done using direct methods (reverse transcription polymerase chain reaction [PCR] and fluorescence in situ hybridization) and indirect methods for ERG mRNA and protein expression using quantitative PCR and immunohistochemistry, respectively. A linear equation method was used to quantitatively determine relative proportions of ERG variants (ERG72/Δ72, ERG81/Δ81) for each sample. RESULTS ERG mRNA and protein expression is increased in patients with advanced prostate cancer, with higher levels of ERG expression significantly associated with seminal vesicle invasion (stage pT3b) and biochemical recurrence. Genes involved in cell migration and invasiveness (matrix metalloproteinase 7, osteopontin, and septin 9) are increased in prostate cancers that overexpress ERG. In addition, there is a clear indication of increased retention of exons 10 and 11 in prostate cancer. CONCLUSIONS Analysis of ERG and its variants may be valuable in determining prognosis and development of prostate cancer.

[1]  R. Shah.  Clinical Applications of Novel ERG Immunohistochemistry in Prostate Cancer Diagnosis and Management , 2013, Advances in anatomic pathology.

[2]  A. Chinnaiyan,et al.  Antibody-based detection of ERG rearrangements in prostate core biopsies, including diagnostically challenging cases: ERG staining in prostate core biopsies. , 2012, Archives of pathology & laboratory medicine.

[3]  Katelyn Powell,et al.  TMPRSS2-ERG Fusion Gene Expression in Prostate Tumor Cells and Its Clinical and Biological Significance in Prostate Cancer Progression. , 2012, Journal of cancer science & therapy.

[4]  A. Villers,et al.  Abnormal Expression of the ERG Transcription Factor in Prostate Cancer Cells Activates Osteopontin , 2011, Molecular Cancer Research.

[5]  O. Kallioniemi,et al.  FZD4 as a mediator of ERG oncogene-induced WNT signaling and epithelial-to-mesenchymal transition in human prostate cancer cells. , 2010, Cancer research.

[6]  A. Vidal-Puig,et al.  LEM-PCR: a method for determining relative transcript isoform proportions using real-time PCR without a standard curve. , 2010, Genome.

[7]  S. Varambally,et al.  Antibody-based detection of ERG rearrangement-positive prostate cancer. , 2010, Neoplasia.

[8]  M. Rubin,et al.  Prevalence of TMPRSS2-ERG and SLC45A3-ERG gene fusions in a large prostatectomy cohort , 2009, Modern Pathology.

[9]  M. Gerstein,et al.  N-myc downstream regulated gene 1 (NDRG1) is fused to ERG in prostate cancer. , 2009, Neoplasia.

[10]  M. Ittmann,et al.  Pleiotropic biological activities of alternatively spliced TMPRSS2/ERG fusion gene transcripts. , 2008, Cancer research.

[11]  S. Dhanasekaran,et al.  A fluorescence in situ hybridization screen for E26 transformation-specific aberrations: identification of DDX5-ETV4 fusion protein in prostate cancer. , 2008, Cancer research.

[12]  Chen Sun,et al.  Delineation of TMPRSS2-ERG Splice Variants in Prostate Cancer , 2008, Clinical Cancer Research.

[13]  T. Tammela,et al.  TMPRSS2:ERG Fusion Identifies a Subgroup of Prostate Cancers with a Favorable Prognosis , 2008, Clinical Cancer Research.

[14]  S. Dhanasekaran,et al.  Distinct classes of chromosomal rearrangements create oncogenic ETS gene fusions in prostate cancer , 2007, Nature.

[15]  Y Pawitan,et al.  TMPRSS2:ERG gene fusion associated with lethal prostate cancer in a watchful waiting cohort , 2007, Oncogene.

[16]  R. Eeles,et al.  Diversity of TMPRSS2-ERG fusion transcripts in the human prostate , 2007, Oncogene.

[17]  Anthony V D'Amico,et al.  Variation in the definition of biochemical recurrence in patients treated for localized prostate cancer: the American Urological Association Prostate Guidelines for Localized Prostate Cancer Update Panel report and recommendations for a standard in the reporting of surgical outcomes. , 2007, The Journal of urology.

[18]  G. Jenster,et al.  TMPRSS2:ERG fusion by translocation or interstitial deletion is highly relevant in androgen-dependent prostate cancer, but is bypassed in late-stage androgen receptor-negative prostate cancer. , 2006, Cancer research.

[19]  O. Kallioniemi,et al.  TMPRSS2 fusions with oncogenic ETS factors in prostate cancer involve unbalanced genomic rearrangements and are associated with HDAC1 and epigenetic reprogramming. , 2006, Cancer research.

[20]  R. Henrique,et al.  TMPRSS2-ERG gene fusion causing ERG overexpression precedes chromosome copy number changes in prostate carcinomas and paired HGPIN lesions. , 2006, Neoplasia.

[21]  Michael Ittmann,et al.  Expression of variant TMPRSS2/ERG fusion messenger RNAs is associated with aggressive prostate cancer. , 2006, Cancer research.

[22]  I. Panagopoulos,et al.  Confirmation of the high frequency of the TMPRSS2/ERG fusion gene in prostate cancer. , 2006, Genes, chromosomes & cancer.

[23]  I. Panagopoulos,et al.  Confirmation of the high frequency of the TMPRSS2/ERG fusion gene in prostate cancer , 2006 .

[24]  V. Srikantan,et al.  Frequent overexpression of ETS-related gene-1 (ERG1) in prostate cancer transcriptome , 2006, Oncogene.

[25]  J. Tchinda,et al.  Recurrent fusion of TMPRSS2 and ETS transcription factor genes in prostate cancer. , 2006, Science.

[26]  V. Srikantan,et al.  Frequent overexpression of ETS-related gene-1 (ERG1) in prostate cancer transcriptome , 2005, Oncogene.

[27]  Tsuneyuki Oikawa,et al.  ETS transcription factors: Possible targets for cancer therapy , 2004, Cancer science.

[28]  C. Caldas,et al.  A simple and reliable pretreatment protocol facilitates fluorescent in situ hybridisation on tissue microarrays of paraffin wax embedded tumour samples , 2003, Molecular pathology : MP.

[29]  K. Grigor,et al.  Androgen receptor gene amplification and protein expression in hormone refractory prostate cancer , 2003, British Journal of Cancer.

[30]  J. Cheville,et al.  Transcriptional silencing of zinc finger protein 185 identified by expression profiling is associated with prostate cancer progression. , 2003, Cancer research.

[31]  J. Rosenbloom,et al.  Transcription Factor Erg Variants and Functional Diversification of Chondrocytes during Limb Long Bone Development , 2000, The Journal of cell biology.

[32]  D. Stéhelin,et al.  Erg proteins, transcription factors of the Ets family, form homo, heterodimers and ternary complexes via two distinct domains , 1998, Oncogene.

[33]  Gil Ast,et al.  Insights into the connection between cancer and alternative splicing. , 2008, Trends in genetics : TIG.

[34]  R. Eeles,et al.  Detection of TMPRSS2-ERG translocations in human prostate cancer by expression profiling using GeneChip Human Exon 1.0 ST arrays. , 2008, The Journal of molecular diagnostics : JMD.