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Title: Identification of multipotent mesenchymal stromal cells in the reactive stroma of a prostate cancer xenograft by side population analysis

Abstract

Cancer stem cells are a distinct cellular population that is believed to be responsible for tumor initiation and maintenance. Recent data suggest that solid tumors also contain another type of stem cells, the mesenchymal stem cells or multipotent mesenchymal stromal cells (MSCs), which contribute to the formation of tumor-associated stroma. The Hoechst 33342 efflux assay has proved useful to identify a rare cellular fraction, named Side Population (SP), enriched in cells with stem-like properties. Using this assay, we identified SP cells in a prostate cancer xenograft containing human prostate cancer cells and mouse stromal cells. The SP isolation, subculture and sequential sorting allowed the generation of single-cell-derived clones of murine origin that were recognized as MSC by their morphology, plastic adherence, proliferative potential, adipogenic and osteogenic differentiation ability and immunophenotype (CD45{sup -}, CD81{sup +} and Sca-1{sup +}). We also demonstrated that SP clonal cells secrete transforming growth factor {beta}1 (TGF-{beta}1) and that their inhibition reduces proliferation and accelerates differentiation. These results reveal the existence of SP cells in the stroma of a cancer xenograft, and provide evidence supporting their MSC nature and the role of TGF-{beta}1 in maintaining their proliferation and undifferentiated status. Our data also reveal the usefulness ofmore » the SP assay to identify and isolate MSC cells from carcinomas.« less

Authors:
 [1];  [2];  [1];  [2];  [2]; ; ;  [1];  [2];  [1];  [2];  [2];  [3];  [4];  [5];  [2];  [1];  [2];  [1];  [2]
  1. Institut de Recerca Hospital Vall d'Hebron, Barcelona (Spain)
  2. (Spain)
  3. (ICREA), Barcelona (Spain)
  4. Institut Curie, Paris (France)
  5. Universitat Autonoma de Barcelona, Barcelona (Spain)
Publication Date:
OSTI Identifier:
22209831
Resource Type:
Journal Article
Resource Relation:
Journal Name: Experimental Cell Research; Journal Volume: 315; Journal Issue: 17; Other Information: Copyright (c) 2009 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; CARCINOMAS; CELL PROLIFERATION; GROWTH FACTORS; INHIBITION; MICE; MORPHOLOGY; PROSTATE; STEM CELLS

Citation Formats

Santamaria-Martinez, Albert, Universitat de Barcelona, Barcelona, Barquinero, Jordi, Universitat Autonoma de Barcelona, Barcelona, Banc de Sang i Teixits, Barcelona, Barbosa-Desongles, Anna, Hurtado, Antoni, Pinos, Tomas, Universitat Autonoma de Barcelona, Barcelona, Seoane, Joan, Universitat Autonoma de Barcelona, Barcelona, Medical Oncology program, Vall d'Hebron Institute of Oncology, Barcelona, Institucio Catalana de Recerca i Estudis Avancats, Poupon, Marie-France, Morote, Joan, Servei d'Urologia. Hospital Vall d'Hebron, Barcelona, Reventos, Jaume, Universitat Autonoma de Barcelona, Barcelona, Munell, Francina, E-mail: fmunell@ir.vhebron.net, and Universitat Autonoma de Barcelona, Barcelona. Identification of multipotent mesenchymal stromal cells in the reactive stroma of a prostate cancer xenograft by side population analysis. United States: N. p., 2009. Web. doi:10.1016/J.YEXCR.2009.05.007.
Santamaria-Martinez, Albert, Universitat de Barcelona, Barcelona, Barquinero, Jordi, Universitat Autonoma de Barcelona, Barcelona, Banc de Sang i Teixits, Barcelona, Barbosa-Desongles, Anna, Hurtado, Antoni, Pinos, Tomas, Universitat Autonoma de Barcelona, Barcelona, Seoane, Joan, Universitat Autonoma de Barcelona, Barcelona, Medical Oncology program, Vall d'Hebron Institute of Oncology, Barcelona, Institucio Catalana de Recerca i Estudis Avancats, Poupon, Marie-France, Morote, Joan, Servei d'Urologia. Hospital Vall d'Hebron, Barcelona, Reventos, Jaume, Universitat Autonoma de Barcelona, Barcelona, Munell, Francina, E-mail: fmunell@ir.vhebron.net, & Universitat Autonoma de Barcelona, Barcelona. Identification of multipotent mesenchymal stromal cells in the reactive stroma of a prostate cancer xenograft by side population analysis. United States. doi:10.1016/J.YEXCR.2009.05.007.
Santamaria-Martinez, Albert, Universitat de Barcelona, Barcelona, Barquinero, Jordi, Universitat Autonoma de Barcelona, Barcelona, Banc de Sang i Teixits, Barcelona, Barbosa-Desongles, Anna, Hurtado, Antoni, Pinos, Tomas, Universitat Autonoma de Barcelona, Barcelona, Seoane, Joan, Universitat Autonoma de Barcelona, Barcelona, Medical Oncology program, Vall d'Hebron Institute of Oncology, Barcelona, Institucio Catalana de Recerca i Estudis Avancats, Poupon, Marie-France, Morote, Joan, Servei d'Urologia. Hospital Vall d'Hebron, Barcelona, Reventos, Jaume, Universitat Autonoma de Barcelona, Barcelona, Munell, Francina, E-mail: fmunell@ir.vhebron.net, and Universitat Autonoma de Barcelona, Barcelona. 2009. "Identification of multipotent mesenchymal stromal cells in the reactive stroma of a prostate cancer xenograft by side population analysis". United States. doi:10.1016/J.YEXCR.2009.05.007.
@article{osti_22209831,
title = {Identification of multipotent mesenchymal stromal cells in the reactive stroma of a prostate cancer xenograft by side population analysis},
author = {Santamaria-Martinez, Albert and Universitat de Barcelona, Barcelona and Barquinero, Jordi and Universitat Autonoma de Barcelona, Barcelona and Banc de Sang i Teixits, Barcelona and Barbosa-Desongles, Anna and Hurtado, Antoni and Pinos, Tomas and Universitat Autonoma de Barcelona, Barcelona and Seoane, Joan and Universitat Autonoma de Barcelona, Barcelona and Medical Oncology program, Vall d'Hebron Institute of Oncology, Barcelona and Institucio Catalana de Recerca i Estudis Avancats and Poupon, Marie-France and Morote, Joan and Servei d'Urologia. Hospital Vall d'Hebron, Barcelona and Reventos, Jaume and Universitat Autonoma de Barcelona, Barcelona and Munell, Francina, E-mail: fmunell@ir.vhebron.net and Universitat Autonoma de Barcelona, Barcelona},
abstractNote = {Cancer stem cells are a distinct cellular population that is believed to be responsible for tumor initiation and maintenance. Recent data suggest that solid tumors also contain another type of stem cells, the mesenchymal stem cells or multipotent mesenchymal stromal cells (MSCs), which contribute to the formation of tumor-associated stroma. The Hoechst 33342 efflux assay has proved useful to identify a rare cellular fraction, named Side Population (SP), enriched in cells with stem-like properties. Using this assay, we identified SP cells in a prostate cancer xenograft containing human prostate cancer cells and mouse stromal cells. The SP isolation, subculture and sequential sorting allowed the generation of single-cell-derived clones of murine origin that were recognized as MSC by their morphology, plastic adherence, proliferative potential, adipogenic and osteogenic differentiation ability and immunophenotype (CD45{sup -}, CD81{sup +} and Sca-1{sup +}). We also demonstrated that SP clonal cells secrete transforming growth factor {beta}1 (TGF-{beta}1) and that their inhibition reduces proliferation and accelerates differentiation. These results reveal the existence of SP cells in the stroma of a cancer xenograft, and provide evidence supporting their MSC nature and the role of TGF-{beta}1 in maintaining their proliferation and undifferentiated status. Our data also reveal the usefulness of the SP assay to identify and isolate MSC cells from carcinomas.},
doi = {10.1016/J.YEXCR.2009.05.007},
journal = {Experimental Cell Research},
number = 17,
volume = 315,
place = {United States},
year = 2009,
month =
}
  • The discovery that the regenerative properties of bone marrow multipotent mesenchymal stromal cells (BM-MSCs) could collaterally favor neoplastic progression has led to a great interest in the function of these cells in tumors. However, the effect of BM-MSCs on colonization, a rate-limiting step of the metastatic cascade, is unknown. In this study, we investigated the effect of BM-MSCs on metastatic outgrowth of B16-F10 melanoma cells. In in vitro experiments, direct co-culture assays demonstrated that BM-MSCs stimulated the proliferation of B16-F10 cells in a dose-dependent manner. For in vivo experiments, luciferase-expressing B16-F10 cells were injected through tail vein and mice weremore » subsequently treated with four systemic injections of BM-MSCs. In vivo bioluminescent imaging during 16 days demonstrated that BM-MSCs enhanced the colonization of lungs by B16-F10 cells, which correlated with a 2-fold increase in the number of metastatic foci. Flow cytometry analysis of lungs demonstrated that although mice harboring B16-F10 metastases displayed more endothelial cells, CD4 T and CD8 T lymphocytes in the lungs in comparison to metastases-free mice, BM-MSCs did not alter the number of these cells. Interestingly, BM-MSCs inoculation resulted in a 2-fold increase in the number of CD11b{sup +} myeloid cells in the lungs of melanoma-bearing animals, a cell population previously described to organize “premetastatic niches” in experimental models. These findings indicate that BM-MSCs provide support to B16-F10 cells to overcome the constraints that limit metastatic outgrowth and that these effects might involve the interplay between BM-MSCs, CD11b{sup +} myeloid cells and tumor cells. - Highlights: • BM-MSCs enhanced B16-F10 proliferation in a dose-dependent manner in vitro. • BM-MSCs facilitated lung colonization by B16-F10 melanoma cells. • BM-MSCs administration did not alter the number of endothelial cells and T lymphocytes in the lungs. • BM-MSCs enhanced the recruitment of CD11b{sup +} myeloid cells during tumor colonization.« less
  • Studies on the role of multipotent mesenchymal stromal cells (MSC) on tumor growth have reported both a tumor promoting and a suppressive effect. The aim of the present study was to determine the effect of MSC isolated from Wharton's jelly of umbilical cord (WJMSC) on lung cancer stem cells (LCSC) derived from human lung tumors: two adenocarcinomas (AC) and two squamous cell carcinomas (SCC). LCSC derived from SCC and AC expressed, to varying extents, the more relevant stem cell markers. The effect of WJMSC on LCSC was investigated in vitro using conditioned medium (WJ-CM): a proliferation increase in AC-LCSC wasmore » observed, with an increase in the ALDH+ and in the CD133+ cell population. By contrast, WJ-CM hampered the growth of SCC-LCSC, with an increase in the pre-G1 phase indicating the induction of apoptosis. Furthermore, the ALDH+ and CD133+ population was also reduced. In vivo, subcutaneous co-transplantation of AC-LCSC/WJMSC generated larger tumors than AC-LCSC alone, characterized by an increased percentage of CD133+ and CD166+ cells. By contrast, co-transplantation of WJMSC and SCC-LCSC did not affect the tumor size. Our results strongly suggest that WJMSC exert, both in vitro and in vivo, contrasting effects on LCSC derived from different lung tumor subtypes. - Highlights: • CM from WJMSC induces apoptosis of SCC-LCSC and reduction of ALDH+ and CD133+ cells. • Specificity of SCC-LCSC inhibition by WJ-CM is proved by the use of a CM from NHDF. • WJ-CM enhance AC-LCSC proliferation and increase CD133+ and ALDH+ cell fractions. • Coinjection of WJMSC with AC-LCSC increase tumor growth with SCC-LCSC has no effect.« less
  • Prostate cancer (CaP) bone metastasis is an early event that remains inactive until later-stage progression. Reduced levels of circulating androgens, due to andropause or androgen deprivation therapies, alter androgen receptor (AR) coactivator expression. Coactivators shift the balance towards enhanced AR-mediated gene transcription that promotes progression to androgen-resistance. Disruptions in coregulators may represent a molecular switch that reactivates latent bone metastasis. Changes in AR-mediated transcription in androgen-sensitive LNCaP and androgen-resistant C4-2 cells were analyzed for AR coregulator recruitment in co-culture with Saos-2 and THP-1. The Saos-2 cell line derived from human osteosarcoma and THP-1 cell line representing human monocytes were usedmore » to display osteoblast and osteoclast activity. Increased AR activity in androgen-resistant C4-2 was due to increased AR expression and SRC1/TIF2 recruitment and decreased SMRT/NCoR expression. AR activity in both cell types was decreased over 90% when co-cultured with Saos-2 or THP-1 due to dissociation of AR from the SRC1/TIF2 and SMRT/NCoR coregulators complex, in a ligand-dependent and cell-type specific manner. In the absence of androgens, Saos-2 decreased while THP-1 increased proliferation of LNCaP cells. In contrast, both Saos-2 and THP-1 decreased proliferation of C4-2 in absence and presence of androgens. Global changes in gene expression from both CaP cell lines identified potential cell cycle and androgen regulated genes as mechanisms for changes in cell proliferation and AR-mediated transactivation in the context of bone marrow stroma cells. - Highlights: • Decreased corepressor expression change AR in androgen-resistance prostate cancer. • Bone stroma-derived cells change AR coregulator recruitment in prostate cancer. • Bone stroma cells change cell proliferation in androgen-resistant cancer cells. • Global gene expression in CaP cells is modified by bone stroma cells in co-cultures. • Potential new multi-subunit coactivator complexes for AR in CaP bone metastasis.« less
  • The role of tumor stroma in regulation of breast cancer growth has been widely studied. However, the details on the type of heterocellular cross-talk between stromal and breast cancer cells (BCCs) are still poorly known. In the present study, in order to investigate the intercellular communication between human mesenchymal stromal cells (hMSCs) and breast cancer cells (BCCs, MDA-MB-231), we recruited cell-internalizing quantum dots (i-QD) generated by conjugation of cell-internalizing anti-mortalin antibody and quantum dots (QD). Co-culture of illuminated and color-coded hMSCs (QD655) and BCCs (QD585) revealed the intercellular transfer of QD655 signal from hMSCs to BCCs. The amount of QDmore » double positive BCCs increased gradually within 48 h of co-culture. We found prominent intercellular transfer of QD655 in hanging drop co-culture system and it was non-existent when hMSCs and BBCs cells were co-cultured in trans-well system lacking imminent cell–cell contact. Fluorescent and electron microscope analyses also supported that the direct cell-to-cell interactions may be required for the intercellular transfer of QD655 from hMSCs to BCCs. To the best of our knowledge, the study provides a first demonstration of transcellular crosstalk between stromal cells and BCCs that involve direct contact and may also include a transfer of mortalin, an anti-apoptotic and growth-promoting factor enriched in cancer cells.« less
  • Purpose: We have previously shown that sodium selenite (SSE) increases radiation-induced cell killing of human prostate carcinoma cells in vitro. In this study we further evaluated the in vivo radiosensitizing effect of SSE in prostate cancer xenograft tumors and normal radiosensitive intestinal crypt cells. Methods and Materials: Immunodeficient (SCID) mice with hormone-independent LAPC-4 (HI-LAPC-4) and PC-3 xenograft tumors (approximately 200 mm{sup 3}) were divided into four groups: control (untreated), radiation therapy (XRT, local irradiation), SSE (2 mg/kg, intraperitoneally, 3 times/week), and XRT plus SSE. The XRT was given at the beginning of the regimen as a single dose of 5more » Gy for HI-LAPC-4 tumors and a single dose of 7 Gy followed by a fractional dose of 3 Gy/d for 5 days for PC-3 tumors. The tumor volume was measured 3 times per week. The radiosensitizing effect of SSE on normal intestinal epithelial cells was assessed by use of a crypt cell microcolony assay. Results: In the efficacy study, SSE alone significantly inhibited the tumor growth in HI-LAPC-4 tumors but not PC-3 tumors. Sodium selenite significantly enhanced the XRT-induced tumor growth inhibition in both HI-LAPC-4 and PC-3 tumors. In the toxicity study, SSE did not affect the intestinal crypt cell survival either alone or in combination with XRT. Conclusions: Sodium selenite significantly enhances the effect of radiation on well-established hormone-independent prostate tumors and does not sensitize the intestinal epithelial cells to radiation. These results suggest that SSE may increase the therapeutic index of XRT for the treatment of prostate cancer.« less