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Extracellular vesicles derived from renal cancer stem cells induce a pro-tumorigenic phenotype in mesenchymal stromal cells.

Lindoso RS, Collino F, Camussi G - Oncotarget (2015)

Bottom Line: We found that CSC-derived EVs promoted persistent phenotypical changes in MSCs characterized by an increased expression of genes associated with cell migration (CXCR4, CXCR7), matrix remodeling (COL4A3), angiogenesis and tumor growth (IL-8, Osteopontin and Myeloperoxidase).Moreover, EV-stimulated MSCs enhanced migration of renal tumor cells and induced vessel-like formation.In conclusion, CSC-derived EVs induced phenotypical changes in MSCs that are associated with tumor growth.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Sciences and Molecular Biotechnology Center University of Torino, Torino, Italy.

ABSTRACT
Renal carcinomas have been shown to contain a population of cancer stem cells (CSCs) that present self-renewing capacity and support tumor growth and metastasis. CSCs were shown to secrete large amount of extracellular vesicles (EVs) that can transfer several molecules (proteins, lipids and nucleic acids) and induce epigenetic changes in target cells. Mesenchymal Stromal Cells (MSCs) are susceptible to tumor signalling and can be recruited to tumor regions. The precise role of MSCs in tumor development is still under debate since both pro- and anti-tumorigenic effects have been reported. In this study we analysed the participation of renal CSC-derived EVs in the interaction between tumor and MSCs. We found that CSC-derived EVs promoted persistent phenotypical changes in MSCs characterized by an increased expression of genes associated with cell migration (CXCR4, CXCR7), matrix remodeling (COL4A3), angiogenesis and tumor growth (IL-8, Osteopontin and Myeloperoxidase). EV-stimulated MSCs exhibited in vitro an enhancement of migration toward the tumor conditioned medium. Moreover, EV-stimulated MSCs enhanced migration of renal tumor cells and induced vessel-like formation. In vivo, EV-stimulated MSCs supported tumor development and vascularization, when co-injected with renal tumor cells. In conclusion, CSC-derived EVs induced phenotypical changes in MSCs that are associated with tumor growth.

No MeSH data available.


Related in: MedlinePlus

Alteration in cytokine secretion by MSCs after CSC-EV stimulation(A) representative cytokine array performed with supernatant of non-stimulated MSCs (lower panel) and stimulated MSCs (upper panel). The three molecules that presented a consistent change in all experiments performed are identified by black box (1-Myeloperoxidase; 2-Osteopontin; 3-IL-8). (B) relative quantification of the spots of the three secreted cytokines. White bars indicate non-stimulated MSC supernatants and black bars represent stimulated MSC supernatants. (C) gene expression of myeloperoxidase, osteopontin and IL-8 in 2 weeks stimulated MSCs (black bars). Non-stimulated MSCs were used as control (white bars). Data are expressed as RQ, normalized to GAPDH. Statistical analysis was performed by t-test: * indicates statistical difference to the control group (P < 0.05; n = 3).
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Figure 5: Alteration in cytokine secretion by MSCs after CSC-EV stimulation(A) representative cytokine array performed with supernatant of non-stimulated MSCs (lower panel) and stimulated MSCs (upper panel). The three molecules that presented a consistent change in all experiments performed are identified by black box (1-Myeloperoxidase; 2-Osteopontin; 3-IL-8). (B) relative quantification of the spots of the three secreted cytokines. White bars indicate non-stimulated MSC supernatants and black bars represent stimulated MSC supernatants. (C) gene expression of myeloperoxidase, osteopontin and IL-8 in 2 weeks stimulated MSCs (black bars). Non-stimulated MSCs were used as control (white bars). Data are expressed as RQ, normalized to GAPDH. Statistical analysis was performed by t-test: * indicates statistical difference to the control group (P < 0.05; n = 3).

Mentions: Cytokine array analysis was performed in cell free supernatants of unstimulated and 2 week-stimulated MSCs to observe variation in the secreted molecules (Fig. 5A and 5B). From the group of cytokines evaluated, three molecules were consistently increased in the supernatant of stimulated MSCs (black bars) in all the experiments performed: IL-8, Myeloperoxidase (Myelop) and Osteopontin (Osteop). To determine if the increase in the secretion of these cytokines was due to a modulation at transcriptional level, we performed gene expression experiments. The results showed that the enhancement in the cytokine release by CSC-EV-stimulated MSCs (black bars) was due to an increase in their gene transcription (Fig. 5C).


Extracellular vesicles derived from renal cancer stem cells induce a pro-tumorigenic phenotype in mesenchymal stromal cells.

Lindoso RS, Collino F, Camussi G - Oncotarget (2015)

Alteration in cytokine secretion by MSCs after CSC-EV stimulation(A) representative cytokine array performed with supernatant of non-stimulated MSCs (lower panel) and stimulated MSCs (upper panel). The three molecules that presented a consistent change in all experiments performed are identified by black box (1-Myeloperoxidase; 2-Osteopontin; 3-IL-8). (B) relative quantification of the spots of the three secreted cytokines. White bars indicate non-stimulated MSC supernatants and black bars represent stimulated MSC supernatants. (C) gene expression of myeloperoxidase, osteopontin and IL-8 in 2 weeks stimulated MSCs (black bars). Non-stimulated MSCs were used as control (white bars). Data are expressed as RQ, normalized to GAPDH. Statistical analysis was performed by t-test: * indicates statistical difference to the control group (P < 0.05; n = 3).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4480728&req=5

Figure 5: Alteration in cytokine secretion by MSCs after CSC-EV stimulation(A) representative cytokine array performed with supernatant of non-stimulated MSCs (lower panel) and stimulated MSCs (upper panel). The three molecules that presented a consistent change in all experiments performed are identified by black box (1-Myeloperoxidase; 2-Osteopontin; 3-IL-8). (B) relative quantification of the spots of the three secreted cytokines. White bars indicate non-stimulated MSC supernatants and black bars represent stimulated MSC supernatants. (C) gene expression of myeloperoxidase, osteopontin and IL-8 in 2 weeks stimulated MSCs (black bars). Non-stimulated MSCs were used as control (white bars). Data are expressed as RQ, normalized to GAPDH. Statistical analysis was performed by t-test: * indicates statistical difference to the control group (P < 0.05; n = 3).
Mentions: Cytokine array analysis was performed in cell free supernatants of unstimulated and 2 week-stimulated MSCs to observe variation in the secreted molecules (Fig. 5A and 5B). From the group of cytokines evaluated, three molecules were consistently increased in the supernatant of stimulated MSCs (black bars) in all the experiments performed: IL-8, Myeloperoxidase (Myelop) and Osteopontin (Osteop). To determine if the increase in the secretion of these cytokines was due to a modulation at transcriptional level, we performed gene expression experiments. The results showed that the enhancement in the cytokine release by CSC-EV-stimulated MSCs (black bars) was due to an increase in their gene transcription (Fig. 5C).

Bottom Line: We found that CSC-derived EVs promoted persistent phenotypical changes in MSCs characterized by an increased expression of genes associated with cell migration (CXCR4, CXCR7), matrix remodeling (COL4A3), angiogenesis and tumor growth (IL-8, Osteopontin and Myeloperoxidase).Moreover, EV-stimulated MSCs enhanced migration of renal tumor cells and induced vessel-like formation.In conclusion, CSC-derived EVs induced phenotypical changes in MSCs that are associated with tumor growth.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Sciences and Molecular Biotechnology Center University of Torino, Torino, Italy.

ABSTRACT
Renal carcinomas have been shown to contain a population of cancer stem cells (CSCs) that present self-renewing capacity and support tumor growth and metastasis. CSCs were shown to secrete large amount of extracellular vesicles (EVs) that can transfer several molecules (proteins, lipids and nucleic acids) and induce epigenetic changes in target cells. Mesenchymal Stromal Cells (MSCs) are susceptible to tumor signalling and can be recruited to tumor regions. The precise role of MSCs in tumor development is still under debate since both pro- and anti-tumorigenic effects have been reported. In this study we analysed the participation of renal CSC-derived EVs in the interaction between tumor and MSCs. We found that CSC-derived EVs promoted persistent phenotypical changes in MSCs characterized by an increased expression of genes associated with cell migration (CXCR4, CXCR7), matrix remodeling (COL4A3), angiogenesis and tumor growth (IL-8, Osteopontin and Myeloperoxidase). EV-stimulated MSCs exhibited in vitro an enhancement of migration toward the tumor conditioned medium. Moreover, EV-stimulated MSCs enhanced migration of renal tumor cells and induced vessel-like formation. In vivo, EV-stimulated MSCs supported tumor development and vascularization, when co-injected with renal tumor cells. In conclusion, CSC-derived EVs induced phenotypical changes in MSCs that are associated with tumor growth.

No MeSH data available.


Related in: MedlinePlus