[1549] Holoclone and Non-Holoclone Derived Cell Lineage miRNA Analysis in Prostate Cancer

YM Salley, P Smyth, CM Martin, O Sheils, JJ O'Leary. Trinity College, Dublin 2, Ireland; Coombe Women and Infants University Hospital, Dublin 8, Ireland

Background: Prostate cancer is the second leading cause of cancer deaths in men. Stem-like cells have been identified in several malignancies including prostate cancer and are thought to drive primary tumorigenesis through self-renewal and differentiation. Additionally, persistence of stem cells post-therapeutic intervention has been proposed as an explanation for metastasis and recurrence. Holoclones are a tightly packed clone of small cells generally thought to contain stem cells and progenitors. MicroRNAs (miRNAs) are recently discovered small family of regulatory molecules that are associated with various malignancies. The aim of this study was to identify a profile of prostate cancer-associated miRNAs, and to derive holoclones from prostate cancer cell lines and to characterise the miRNA profile of these stem-like cells.
Design: In this study, meta-analysis was carried out to compare existing data on miRNA expression in prostate cancer and data was analysed using miRGen and miRNApath. The expression of a panel of known human miRNAs, was assessed in a group of prostate cell lines PWR-1E (normal), LNCaP (metastatic carcinoma), PC-3 (non-metastatic adenocarcinoma) using a quantitative Real Time TaqMan PCR method.
Results: Meta-analysis identified common miRNA species in prostate cancer and the predicted gene targets and pathways were also assessed. Putative holoclones were generated from cell lines (LNCaP, PC-3) using a high salt-soft agar assay and LNCaP putative holoclones were kept alive for 24 days and PC-3 holoclones were kept alive for 6 days (LNCaP cell line represents a metastatic prostatic carcinoma and should contain a higher number of cancer stem cells). miRNA profiling was performed using 380 individual assays based on Stem Looped Primer PCR reactions.
Conclusions: Analysis of the data showed unique miRNA populations varied between each of the cells profiled. Future work will consist of further analysis on the expression of human miRNAs in prostate cancer cell lines and holoclone derived stem cells in order to identify prostate cancer stem cell-specific miRNA populations. Predicted gene targets and pathways will also be assessed in the cell lines and holoclones, and then compared to the meta-analysis study. Acknowlegements: Prostate Cancer Research Consortium CERVIVA (ICSRC)
Category: Pathobiology

Tuesday, March 10, 2009 1:00 PM

Poster Session IV # 206, Tuesday Afternoon