Holoclone and Non-Holoclone Derived Cell Lineage Characterisation Analysis in Prostate Cancer
YM Salley, MF Gallagher, PC Smyth, CM Martin, OM Sheils, JJ O'Leary. Trinity College, Dublin 2, Ireland
Background: Prostate cancer is a heterogeneous disease and is the second most common cause of cancer resulting in male deaths. 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. The aim of this study was to derive holoclones from prostate cancer cell lines and to characterise their expression.
Design: In this study, holoclones were cultured using a high salt-soft agar assay for LNCaP (metastatic carcinoma) and PC-3 (non-metastatic adenocarcinoma) cell lines. The expression of a panel of key stemness genes and pathways, was assessed in a group of prostate holoclones derived from cell lines LNCaP and PC-3 using a quantitative Real Time TaqMan® PCR method.
Results: Holoclones were generated from cell lines (LNCaP, PC-3) using a high salt-soft agar assay. LNCaP holoclones were maintained for 24 days and PC-3 holoclones were maintained for 6 days. The differential duration of the time points denotes that the LNCaP cell line represents a metastatic prostatic carcinoma and should contain a higher number of cancer stem cells. In combination, stemness genes were used to exemplify the stem cell potential and characteristics of these hololcones. Examples of stemness genes include Sonic Hedgehog, TGF-β, Oct4 and Nanog.
Conclusions: Expression profiles were generated for genes representing stemness. LNCaP and PC-3 holoclones both indicated stemness characteristics. However both LNCaP and PC-3 holoclones expressed different stemness markers and this disparity was also further represented by the duration differences in time points. Future work will consist of further characterisation and analysis of LNCaP and PC-3 holoclones. Acknowledgements: Prostate Cancer Research Consortium
Wednesday, March 24, 2010 9:30 AM
Poster Session V # 210, Wednesday Morning