title: Comparative microRNA profiling of prostate carcinomas with increasing tumor stage by deep sequencing.
abstract: MicroRNAs (miRNA) posttranscriptionally regulate gene expression and are important in tumorigenesis. Previous deep sequencing identified the miRNA profile of prostate carcinoma versus nonmalignant prostate tissue. Here, we generated miRNA expression profiles of prostate carcinoma by deep sequencing, with increasing tumor stage relative to corresponding nonmalignant and healthy prostate tissue, and detected clearly changed miRNA expression patterns. The miRNA profiles of the healthy and nonmalignant tissues were consistent with our previous findings, indicating a high fidelity of the method employed. In the tumors, quantitative real-time PCR (qRT-PCR) analysis of 40 paired samples of prostate carcinoma versus normal tissue revealed significant upregulation of miR-20a, miR-148a, miR-200b, and miR-375 and downregulation of miR-143 and miR-145. Hereby, miR-375 increased from normal to organ-confined tumors (pT2 pN0), slightly decreased in tumors with extracapsular growth (pT3 pN0), but was then expressed again at higher levels in lymph node metastasizing (pN1) tumors. The sequencing data for miR-375 were confirmed by Northern blotting and qRT-PCR. The regulation for other selected miRNAs could, however, not be confirmed by qRT-PCR in individual tumor stages. MiR-200b, in addition to miR-200c and miR-375 reduced the expression of SEC23A. Interestingly, miR-375, found by sequencing in pT2 upregulated by us and others in tumor versus normal tissue, and miR-15a, found by sequencing in pT2 and pT3 and in the metastasizing tumors, target the phosphatases PHLPP1 and PHLPP2, respectively. PHLPP1 and PHLPP2 dephosphorylate members of the AKT family of signal transducers, thereby inhibiting cell growth. Coexpression of miR-15a and miR-375 resulted in downregulation of PHLPP1/2 and strongly increased prostate carcinoma cell growth.
  
  
  CONCLUSIONS
  These genomic data reveal relevant miRNAs in prostate cancer that may have biomarker and therapeutic potential.
authors: Saarland University Medical School; Department of Virology, Kirrbergerstrasse, Haus 47, 66421 Homburg/Saar, Germany. Graesser@uks.eu.
authors: Martin
authors: M
authors: Hart
authors: Elke
authors: E
authors: Nolte
authors: Sven
authors: S
authors: Wach
authors: Jaroslaw
authors: J
authors: Szczyrba
authors: Helge
authors: H
authors: Taubert
authors: Tilman T
authors: TT
authors: Rau
authors: Arndt
authors: A
authors: Hartmann
authors: Friedrich A
authors: FA
authors: Grässer
authors: Bernd
authors: B
authors: Wullich
mesh_terms: D000368:Aged; D015415:Biomarkers; D045744:Cell Line, Tumor; D020869:Gene Expression Profiling / Q000379:methods*; D015972:Gene Expression Regulation, Neoplastic; D057809:HEK293 Cells; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D008207:Lymphatic Metastasis / Q000235:genetics* / Q000473:pathology; D008297:Male; D035683:MicroRNAs / Q000235:genetics*; D008875:Middle Aged; D009367:Neoplasm Staging; D011471:Prostatic Neoplasms / Q000235:genetics*
chemical_list: D015415:Biomarkers; D035683:MicroRNAs