Molecular Profiling of Prostatic Acinar Morphogenesis Identifies PDCD4 and KLF6 as Tissue Architecture–Specific Prognostic Markers in Prostate Cancer

• DOI: 10.1016/j.ajpath.2012.10.024
• Publication Date: 2012-12-04T21:45:17Z
• Authors: Chi-Rong Li; Jimmy J.-M. Su; Wei-Yu Wang; Michael T.-L. Lee; Ting-Yun Wang; Kuan-Ying Jiang; Chein-Feng Li; Jong-Ming Hsu; Chi-Kuan Chen; Marcelo Chen; Shih-Sheng Jiang; Valerie M. Weaver; Kelvin K.-C. Tsai
• Keywords: Prostatic Diseases

Histopathological classification of human prostate cancer (PCA) relies on the morphological assessment tissue specimens but has limited prognostic value. To address this deficiency, we performed comparative transcriptome analysis prostatic acini generated in a three-dimensional basement membrane that recapitulates differentiated characteristics and gene expression profile glandular epithelial tissue. We then applied an acinar morphogenesis–specific to two independent cohorts patients with PCA (total n = 79) found those tumors expressing profile, which designated acini-like tumors, had significantly lower risk postoperative relapse compared correlation (hazard ratio, 0.078; log-rank test P 0.009). Multivariate analyses showed superior prediction performance using system clinical criteria Gleason scores. prioritized genes identified programmed cell death protein 4 (PDCD4) Kruppel-like factor 6 (KLF6) as critical regulators surrogate markers architectures, form signature robustly predicts prognosis remarkable accuracy several large series 161; concordance index, 0.913 0.951). Thus, by exploiting genomic program associated differentiation, related molecular enhance PCA. Prostate is leading cause cancer-related men. For early-stage localized cancer, radical prostatectomy offers best opportunity eradicate disease. However, approximately 15% 30% disease at diagnosis develop recurrence within 5 10 years, most these subsequently show poor therapeutic outcome.1Bill-Axelson A. Holmberg L. Ruutu M. Haggman Andersson S.O. Bratell S. Spangberg Busch C. Nordling Garmo H. Palmgren J. Adami H.O. Norlen B.J. Johansson J.E. 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Three-dimensional models cells.Nat Methods. 4: 359-365Crossref (1000) Confocal imaging Nikon Digital Eclipse C1 confocal microscope (Nikon, Tokyo, Japan). antibodies include rat anti-α6 (clone GoH3; Millipore) mouse anti-GM130 35; Biosciences). nuclei counterstained Hoechst 33342 DAPI (Invitrogen). Total RNA samples extracted cultivated 3D rBM different lengths time (36 48 hours 7 8 days) TRIzol (Invitrogen) purified RNeasy minikit DNase (Qiagen, Valencia, CA). Experiments triplicate. Whole-genome Affymetrix Human Genome U133A 2.0 Plus GeneChip platform, protocol (Affymetrix, Santa Clara, hybridization intensity data processed Operating software (Affymetrix), filtered P/A/M flags retain present all three least one conditions. false-discovery rate <0.025 select differentially expressed comparison group. deposited National Center Biotechnology Information's Omnibus (http://www.ncbi.nlm.nih.gov/geo; accession number GSE30304). (1.0 μg) template cDNA synthesis Moloney Murine Leukemia Virus Reverse Transcriptase (Promega, Madison, WI). (100 ng) PCR amplification LightCycler FastStart DNA MASTERPLUS SYBR Green I Kit, quantitative real-time RT-PCR amplified Kit (Roche Diagnostics GmbH, Mannheim, Germany). 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Endogenous peroxidase activity quenched 3% hydrogen 15 minutes, slides incubated horse block non-specific immunoreactivity. antibody detected DAKO EnVision kit (Dako, Carpinteria, anti-PDCD4 (1:4000, clone EPR3431), anti-KLF6 (1:50, polyclonal), anti–ATP-binding cassette transporter (ABCG1; 1:500, EP1366Y) (all Epitomics, Burlingame, All immunohistochemical (IHC) evaluated expert pathologists (C.-F.Li C.-K. Chen), quantified histological (H-score).29Budwit-Novotny D.A. McCarty K.S. E.B. Soper Mutch Creasman W.T. Flowers Immunohistochemical estrogen receptor endometrial adenocarcinoma monoclonal antibody.Cancer 1986; 46: 5419-5425PubMed ScholarTable 1Patient CharacteristicsClinical cohortBWH group (n 50)SU/KI/JHU 29)CFMC 61)MMH 50)Mean (SD) age (years)59.4 (6.8)59.0 (6.2)65.1 (5.6)67.4 (5.4)Gleason 4-746 (92.0)22 (75.9)29 (47.5)20 (40.0) 8-94 (8.0)7 (24.1)32 (52.5)30 (60.0)Mean baseline (ng/mL)8.3 (5.8)NA18.8 (30.8)15.2 (3.6)Tumor stage I-II30 (60.0)19 (65.5)53 (86.9)42 (84.0) ≥III20 (40.0)10 (34.5)8 (13.1)8 (16.0)Recurrence9 (18.0)7 (24.1)11 (18.0)12 (24.0)Medium (months)42.911.587.970.2MMH, Hospital; NA, available. Open table tab MMH, statistical programming language R 2.14.0 (http://cran.r-project.org) SPSS 10.0 (SPSS, Chicago, IL), conduct data. two-tailed Student's t-test simple significance testing, tests analysis. C-statistics conducted survcomp package. cutoff discriminates groups respect maximal Youden's index.30Pepe Statistical Evaluation Tests Classification Prediction. 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