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Volume 69, Issue 4, Pages (April 2016)

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1 Volume 69, Issue 4, Pages 551-554 (April 2016)
Telomerase Activity and Telomere Length in Human Benign Prostatic Hyperplasia Stem- like Cells and Their Progeny Implies the Existence of Distinct Basal and Luminal Cell Lineages  Jayant K. Rane, Sarah Greener, Fiona M. Frame, Vincent M. Mann, Matthew S. Simms, Anne T. Collins, Daniel M. Berney, Norman J. Maitland  European Urology  Volume 69, Issue 4, Pages (April 2016) DOI: /j.eururo Copyright © 2015 European Association of Urology Terms and Conditions

2 Fig. 1 Telomerase biology in epithelial subpopulations derived from human benign prostatic hyperplasia. (a) Quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis for TERC expression (n=5). Data normalised to RPLP0 expression. (b) qRT-PCR analysis for TERT expression (n=5). Data normalised to RPLP0 expression. (c) qRT-PCR analysis to measure telomerase activity using TRAPeze RT telomerase detection kit (Millipore, Bedford, MA, USA) (n=5). Data normalised to telomerase activity in the P4E6 cell line. (d) Relative telomere length determination using modified qRT-PCR method (n=5). The telomere length for each patient sample was normalised to the individual patients’ lymphocyte telomere lengths. Error bars represent mean ± SD. * p<0.05. ** p<0.01. *** p<0.001. BPH=benign prostatic hyperplasia; CB=committed basal cells; HI=heat inactivated control; LC=luminal cells; mRNA=messenger RNA; ND=not detected; SC=stem-like cells; TA=transit-amplifying cells. European Urology  , DOI: ( /j.eururo ) Copyright © 2015 European Association of Urology Terms and Conditions

3 Fig. 2 Distinct basal and luminal hyperproliferation in benign prostatic hyperplasia tissue. (a) Immunofluorescence analysis of formalin-fixed paraffin-embedded sections showing predominantly TP63+ (magenta) basal hyperproliferation (first panel), predominantly NKX3.1+ (green) luminal hyperproliferation (second panel), and simultaneous basal and luminal hyperproliferation (third panel) in patient-derived benign prostatic hyperplasia (BPH) tissues. The arrows indicate areas of hyperproliferation. All nuclei are stained blue (DAPI). (b) A box and whisker plot showing variation of basal to luminal ratios in BPH. The number of basal cells (nuclear TP63 staining) and luminal cells (present in gland, TP63 negative) were counted. Images were captured on a Nikon Eclipse TE300 fluorescent microscope (Nikon, Tokyo, Japan) or Axio Scan.Z1 slide scanner (Carl Zeiss Microscopy GmbH, Jena, Germany). Cells were counted by eye. A total of 2800 cells were counted from 13 patients. (c) Model for prostate epithelial stem cell fate in BPH: The stem-like cell differentiates into basal and luminal progenitors. Alternatively, luminal progenitors can be derived from basal progenitor cells. The luminal progenitor then differentiates into single or very limited luminal cells. The basal progenitor cell gives rise to several basal cells, which retain some proliferative potential. Because these basal cells do not exhibit telomerase activity, they possess shorter telomeres. European Urology  , DOI: ( /j.eururo ) Copyright © 2015 European Association of Urology Terms and Conditions


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