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Invasive prostate cancer cells are tumor initiating cells that have a stem cell-like genomic signature

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Abstract

Development of metastasis is a leading cause of cancer-induced death. Acquisition of an invasive tumor cell phenotype suggests loss of cell adhesion and basement membrane breakdown during a process termed epithelial-to-mesenchymal transition (EMT). Recently, cancer stem cells (CSC) were discovered to mediate solid tumor initiation and progression. Prostate CSCs are a subpopulation of CD44+ cells within the tumor that give rise to differentiated tumor cells and also self-renew. Using both primary and established prostate cancer cell lines, we tested the assumption that CSCs are more invasive. The ability of unsorted cells and CD44-positve and -negative subpopulations to undergo Matrigel invasion and EMT was evaluated, and the gene expression profiles of these cells were analyzed by microarray and a subset confirmed using QRT-PCR. Our data reveal that a subpopulation of CD44+ CSC-like cells invade Matrigel through an EMT, while in contrast, CD44− cells are non-invasive. Furthermore, the genomic profile of the invasive cells closely resembles that of CD44+CD24− prostate CSCs and shows evidence for increased Hedgehog signaling. Finally, invasive cells from DU145 and primary prostate cancer cells are more tumorigenic in NOD/SCID mice compared with non-invasive cells. Our data strongly suggest that basement membrane invasion, an early and necessary step in metastasis development, is mediated by these potential cancer stem cells.

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Acknowledgments

This project has been funded in whole or in part with federal funds from the National Cancer Institute, National Institutes of Health, under contract N01-CO-12400, and supported (in part) by the Intramural Research Program of the NIH, National Cancer Institute, Center for Cancer Research. The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the US Government.

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Authors and Affiliations

  1. Cancer Stem Cell Section, Laboratory of Cancer Prevention, SAIC-Frederick Inc., National Cancer Institute at Frederick, Frederick, MD, 21702, USA

    George J. Klarmann, Xiaohu Zhang & Suneetha B. Thomas

  2. Cancer Stem Cell Section, Laboratory of Cancer Prevention, Center for Cancer Research, National Cancer Institute at Frederick, 1050 Boyles St., Building 560, Room 21-81, Frederick, MD, 21702, USA

    Elaine M. Hurt, Lesley A. Mathews, Maria A. Duhagon, Tashan Mistree & William L. Farrar

  3. LIM-F. Ciencias, Igua 4225, Universidad de la República, CP 11400/Genetica-F. Medicina, Gral. Flores 2125, CP 11800, Montevideo, Uruguay

    Maria A. Duhagon

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  1. George J. Klarmann
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Correspondence to William L. Farrar.

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10585_2009_9242_MOESM1_ESM.eps

Supplemental Fig. 1 QRT-PCR validation analysis of changes in mRNA between top and bottom cells. A. Changes in EMT markers and B. changes in stem cell genes. Error bars represent SEM from two independent samples and were normalized to expression of 18S rRNA. All changes are significantly different from top cells (P < 0.05) with the exception of TGFB1 and vimentin in LNCaP cells. Fold-induction changes for LNCaP cells: 0.355-CD24, 0.308-E-cadherin, 0.733-vimentin, 3.622-BMI1, 3.690-Nanog, 3.355-FGF2 and 1.2 for TGFB1. Fold-induction changes for DU145 cells: 5.104-CD44, 0.104-E-cadherin, 14.028-vimentin, 19.326-BMI1, 2.714-Nanog, 2.049-FGF2, 7.450-TGFB1 and 1.55 for SHH (EPS 1412 kb)

Supplemental Fig. 2 CD44+ stem-cells like do not contain an EMT genotype prior to plating on Matrigel (EPS 867 kb)

Supplementary Table S1 (XLS 1932 kb)

Supplementary Table S2 (XLS 41 kb)

Supplementary Table S3 (XLS 83 kb)

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Klarmann, G.J., Hurt, E.M., Mathews, L.A. et al. Invasive prostate cancer cells are tumor initiating cells that have a stem cell-like genomic signature. Clin Exp Metastasis 26, 433–446 (2009). https://doi.org/10.1007/s10585-009-9242-2

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