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dc.contributor.authorOrr, Brigiden
dc.date.accessioned2018-03-29T12:19:29Z
dc.date.available2018-03-29T12:19:29Z
dc.date.issued2006
dc.identifier.urihttp://hdl.handle.net/1842/29308
dc.description.abstracten
dc.description.abstractGrowth and development of the prostate are regulated by androgens and mesenchymalepithelial interactions. Prostate growth starts in the embryo, continues throughout neonatal life, and is completed at puberty. The male and female embryonic urogenital sinus (UGS) both contain a prostate precursor, but due to the action of testosterone only the male will develop a prostate. Testosterone acts in concert with paracrine signalling from inductive mesenchyme to regulate epithelial growth and branching of the prostate. The ventral mesenchymal pad (VMP) is an area of inductive mesenchyme that is rich in regulatory growth factors of which few have been identified at present. Recently, the role of mesenchyme/stroma in cancer has been established and developmental factors may be involved in mediating the effects of cancer stroma. The aim of this thesis was to identify mesenchymal factors and characterise their expression and function in the developing prostate, and to investigate their expression in prostate cancer stroma.en
dc.description.abstractThe LongSAGE technique was used to generate a comprehensive transcriptional profile of the neonatal rat prostate rudiment. LongSAGE is an open-ended and unbiased geneprofiling method. Two libraries were made from tissues in the female UGS; one library was prepared from the whole female prostate rudiment and the other library was prepared from the inductive mesenchyme (VMP) isolated from within the rudiment. Several essential factors known to be involved in prostate development were identified in the libraries, including low abundance cDNAs such as AR and FgflO. This suggested that the approach had sufficient sensitivity to identify key mesenchymal factors. The two libraries were compared, and the comparison was statistically analysed, highlighting genes that were VMP-enriched (P<0.05). Candidate mesenchymal transcripts were selected from the VSU and VMP Libraries by two different approaches, either because of their status as VMP -enriched (P<0.05) transcripts, or by an 'intuitive' approach, because the transcripts were associated with genes that are known members of developmental pathways and/or have been associated with prostate cancer. The expression and abundance of candidate transcripts were quantified by qRT-PCR in the male and female neonatal UGS. Subsequently, candidate transcripts were verified as VMP-enriched and were quantified during prostate development by Northern blot analysis. The protein distribution of selected candidates were localised within the neonatal rat UGS by immunocytochemistry, and the effect of testosterone treatment on the protein distribution was studied. To test the function of one of the candidates on prostate growth and development, recombinant protein was added to prostate rudiments grown in vitro. Finally, candidate transcripts were investigated in human prostate cancer associated fibroblast cells (CAFs) and normal prostate fibroblast cells (NPFs) by RT-PCR and Northern blot analysis.en
dc.description.abstractSAGE analysis, qRT-PCR and Northen blot analysis identified six candidate transcripts as VMP-enriched; Dlkl, Notch2, Ptn Nell2, MMP2 and MMP14. The transcript expression of each candidate was most abundant in the developing rat prostate during the perinatal period. Proteins for Dlkl, Ptn and Notch2 were localised to mesenchymal cells of the neonatal VMP and ventral prostate (VP). Ptn expression was also associated with the basement membrane and cell-surface of the epithelial duct cells of the VP. Treatment of VP organs with recombinant DLK1 in vitro increased the organ size and epithelial branching. Also, PTN, NOTCH2, MMP2 and MMP14 transcript expression was observed in CAFs and NPFs. PTN and NOTCH2 showed a decrease in CAFs compared to NPFs suggesting a tumour-suppressive role.en
dc.description.abstractIn summary, a comprehensive gene profiling technique was used to identify mesenchyme specific/enriched transcripts in the developing prostate. The expression, distribution and function of candidate transcripts and proteins were investigated in the developing prostate and in prostate cancer stroma.en
dc.publisherThe University of Edinburghen
dc.relation.ispartofAnnexe Thesis Digitisation Project 2018 Block 17en
dc.relation.isreferencedbyAlready catalogueden
dc.titleIdentification of mesenchymal genes involved in prostate organogenesis by LongSAGE analysisen
dc.typeThesis or Dissertationen
dc.type.qualificationlevelDoctoralen
dc.type.qualificationnamePhD Doctor of Philosophyen


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