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Microparticles mediated cross-talk between tumoral and endothelial cells promote the constitution of a pro-metastatic vascular niche through Arf6 up regulation.

Pasquier J, Thawadi HA, Ghiabi P, Abu-Kaoud N, Maleki M, Guerrouahen BS, Vidal F, Courderc B, Ferron G, Martinez A, Al Sulaiti H, Gupta R, Rafii S, Rafii A - Cancer Microenviron (2014)

Bottom Line: Our data show that MPs from mesenchymal-like cell lines (MDA-MB231, SKOV3 and APOCC) were able to promote an activation of endothelial cells through Akt phosphorylation, compared to MPs from epithelial-like cell lines (OVCAR3 and MCF7).The endothelial activation was associated to increased Arf6 expression and MPs secretion.Such cross-talk may play a role in perfusion independent role of the endothelium.

View Article: PubMed Central - PubMed

Affiliation: Stem Cell and Microenvironment Laboratory, Weill Cornell Medical College in Qatar, Education City, Qatar Foundation, Doha, Qatar.

ABSTRACT
The tumor stroma plays an essential role in tumor growth, resistance to therapy and occurrence of metastatic phenotype. Tumor vessels have been considered as passive conducts for nutrients but several studies have demonstrated secretion of pro-tumoral factors by endothelial cells. The failure of anti-angiogenic therapies to meet expectations raised by pre-clinical studies prompt us to better study the cross-talk between endothelial and cancer cells. Here, we hypothesized that tumor cells and the endothelium secrete bio-active microparticles (MPs) participating to a functional cross-talk. We characterized the cancer cells MPs, using breast and ovarian cancer cell lines (MCF7, MDA-MB231, SKOV3, OVCAR3 and a primary cell lines, APOCC). Our data show that MPs from mesenchymal-like cell lines (MDA-MB231, SKOV3 and APOCC) were able to promote an activation of endothelial cells through Akt phosphorylation, compared to MPs from epithelial-like cell lines (OVCAR3 and MCF7). The MPs from mesenchymal-like cells contained increased angiogenic molecules including PDGF, IL8 and angiogenin. The endothelial activation was associated to increased Arf6 expression and MPs secretion. Endothelial activation functionalized an MP dependent pro-tumoral vascular niche promoting cancer cells proliferation, invasiveness, stem cell phenotype and chemoresistance. MPs from cancer and endothelial cells displayed phenotypic heterogeneity, and participated to a functional cross-talk where endothelial activation by cancer MPs resulted in increased secretion of EC-MPs sustaining tumor cells. Such cross-talk may play a role in perfusion independent role of the endothelium.

No MeSH data available.


Related in: MedlinePlus

EMT in cancer cells induce a modification in the microparticles released. a MCF7 were treated with TGFβ (2.5 ng/ml) during 3 days. Phase contrast microscopy showing significant morphological change concordant with a mesenchymal phenotype. Scale bar 500 μm. b After treatment with TFGβ, MCF7 were stained for E-cadherin or N-cadherin and analyzed by confocal microscopy. An increase of N-cadherin staining and a decrease of E-cadherin staining can be observed in the MCF7 treated with TGFβ in comparison to controls. Scale bar: 10 μm. c The relative quantification of EMT genes were performed by real-time qPCR on MCF7 after treatment with TGFβ. The mesenchymal genes (N-cadherin, Snail, Fibronectin and Vimentin) were increased compared to the control. E-cadherin expression was decreased. Relative transcript levels are represented as the log10 of ratios between the 2 subpopulations of their 2–ΔΔCp real-time PCR values. d Schematic representation of the experimental design to assess the functional role of iM-MPs. e HUVECs were plated and counted every 2 days in presence of MCF7-MPs or iM-MPs. Only iM-MPs were able to sustain proliferation of HUVECs. f HUVECs were plated on matrigel layer in presence of MCF7-MPs or iM-MPs. Only iM-MPs were able to improve the number of tube and their viability
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Fig3: EMT in cancer cells induce a modification in the microparticles released. a MCF7 were treated with TGFβ (2.5 ng/ml) during 3 days. Phase contrast microscopy showing significant morphological change concordant with a mesenchymal phenotype. Scale bar 500 μm. b After treatment with TFGβ, MCF7 were stained for E-cadherin or N-cadherin and analyzed by confocal microscopy. An increase of N-cadherin staining and a decrease of E-cadherin staining can be observed in the MCF7 treated with TGFβ in comparison to controls. Scale bar: 10 μm. c The relative quantification of EMT genes were performed by real-time qPCR on MCF7 after treatment with TGFβ. The mesenchymal genes (N-cadherin, Snail, Fibronectin and Vimentin) were increased compared to the control. E-cadherin expression was decreased. Relative transcript levels are represented as the log10 of ratios between the 2 subpopulations of their 2–ΔΔCp real-time PCR values. d Schematic representation of the experimental design to assess the functional role of iM-MPs. e HUVECs were plated and counted every 2 days in presence of MCF7-MPs or iM-MPs. Only iM-MPs were able to sustain proliferation of HUVECs. f HUVECs were plated on matrigel layer in presence of MCF7-MPs or iM-MPs. Only iM-MPs were able to improve the number of tube and their viability

Mentions: Our data suggested that M-MPs displayed a specific ability to activate ECs compare to E-MPs. However, this could be cell-specific rather than phenotype-related. Therefore we used an in vitro Epithelial to Mesenchymal Transition (EMT) model using TGFβ to investigate the functional modifications of corresponding MPs [35]. MCF7 cells were treated by TGFβ for 3 days until morphological changes appeared (Fig. 3a). EMT was then confirmed by reduced expression of E-cadherin and increased expression of N-cadherin, Vimentin, Fibronectin and the transcription factor SNAIL (Fig. 3b-c). We subsequently starved the EMT cells and isolated the induced mesenchymal MPs (iM-MPs) (Fig. 3d). We showed that the iM-MPs were able to increase both ECs proliferation and branching in a Matrigel assay, compared to the MCF7-MPs (Fig. 3e-f).Fig. 3


Microparticles mediated cross-talk between tumoral and endothelial cells promote the constitution of a pro-metastatic vascular niche through Arf6 up regulation.

Pasquier J, Thawadi HA, Ghiabi P, Abu-Kaoud N, Maleki M, Guerrouahen BS, Vidal F, Courderc B, Ferron G, Martinez A, Al Sulaiti H, Gupta R, Rafii S, Rafii A - Cancer Microenviron (2014)

EMT in cancer cells induce a modification in the microparticles released. a MCF7 were treated with TGFβ (2.5 ng/ml) during 3 days. Phase contrast microscopy showing significant morphological change concordant with a mesenchymal phenotype. Scale bar 500 μm. b After treatment with TFGβ, MCF7 were stained for E-cadherin or N-cadherin and analyzed by confocal microscopy. An increase of N-cadherin staining and a decrease of E-cadherin staining can be observed in the MCF7 treated with TGFβ in comparison to controls. Scale bar: 10 μm. c The relative quantification of EMT genes were performed by real-time qPCR on MCF7 after treatment with TGFβ. The mesenchymal genes (N-cadherin, Snail, Fibronectin and Vimentin) were increased compared to the control. E-cadherin expression was decreased. Relative transcript levels are represented as the log10 of ratios between the 2 subpopulations of their 2–ΔΔCp real-time PCR values. d Schematic representation of the experimental design to assess the functional role of iM-MPs. e HUVECs were plated and counted every 2 days in presence of MCF7-MPs or iM-MPs. Only iM-MPs were able to sustain proliferation of HUVECs. f HUVECs were plated on matrigel layer in presence of MCF7-MPs or iM-MPs. Only iM-MPs were able to improve the number of tube and their viability
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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Fig3: EMT in cancer cells induce a modification in the microparticles released. a MCF7 were treated with TGFβ (2.5 ng/ml) during 3 days. Phase contrast microscopy showing significant morphological change concordant with a mesenchymal phenotype. Scale bar 500 μm. b After treatment with TFGβ, MCF7 were stained for E-cadherin or N-cadherin and analyzed by confocal microscopy. An increase of N-cadherin staining and a decrease of E-cadherin staining can be observed in the MCF7 treated with TGFβ in comparison to controls. Scale bar: 10 μm. c The relative quantification of EMT genes were performed by real-time qPCR on MCF7 after treatment with TGFβ. The mesenchymal genes (N-cadherin, Snail, Fibronectin and Vimentin) were increased compared to the control. E-cadherin expression was decreased. Relative transcript levels are represented as the log10 of ratios between the 2 subpopulations of their 2–ΔΔCp real-time PCR values. d Schematic representation of the experimental design to assess the functional role of iM-MPs. e HUVECs were plated and counted every 2 days in presence of MCF7-MPs or iM-MPs. Only iM-MPs were able to sustain proliferation of HUVECs. f HUVECs were plated on matrigel layer in presence of MCF7-MPs or iM-MPs. Only iM-MPs were able to improve the number of tube and their viability
Mentions: Our data suggested that M-MPs displayed a specific ability to activate ECs compare to E-MPs. However, this could be cell-specific rather than phenotype-related. Therefore we used an in vitro Epithelial to Mesenchymal Transition (EMT) model using TGFβ to investigate the functional modifications of corresponding MPs [35]. MCF7 cells were treated by TGFβ for 3 days until morphological changes appeared (Fig. 3a). EMT was then confirmed by reduced expression of E-cadherin and increased expression of N-cadherin, Vimentin, Fibronectin and the transcription factor SNAIL (Fig. 3b-c). We subsequently starved the EMT cells and isolated the induced mesenchymal MPs (iM-MPs) (Fig. 3d). We showed that the iM-MPs were able to increase both ECs proliferation and branching in a Matrigel assay, compared to the MCF7-MPs (Fig. 3e-f).Fig. 3

Bottom Line: Our data show that MPs from mesenchymal-like cell lines (MDA-MB231, SKOV3 and APOCC) were able to promote an activation of endothelial cells through Akt phosphorylation, compared to MPs from epithelial-like cell lines (OVCAR3 and MCF7).The endothelial activation was associated to increased Arf6 expression and MPs secretion.Such cross-talk may play a role in perfusion independent role of the endothelium.

View Article: PubMed Central - PubMed

Affiliation: Stem Cell and Microenvironment Laboratory, Weill Cornell Medical College in Qatar, Education City, Qatar Foundation, Doha, Qatar.

ABSTRACT
The tumor stroma plays an essential role in tumor growth, resistance to therapy and occurrence of metastatic phenotype. Tumor vessels have been considered as passive conducts for nutrients but several studies have demonstrated secretion of pro-tumoral factors by endothelial cells. The failure of anti-angiogenic therapies to meet expectations raised by pre-clinical studies prompt us to better study the cross-talk between endothelial and cancer cells. Here, we hypothesized that tumor cells and the endothelium secrete bio-active microparticles (MPs) participating to a functional cross-talk. We characterized the cancer cells MPs, using breast and ovarian cancer cell lines (MCF7, MDA-MB231, SKOV3, OVCAR3 and a primary cell lines, APOCC). Our data show that MPs from mesenchymal-like cell lines (MDA-MB231, SKOV3 and APOCC) were able to promote an activation of endothelial cells through Akt phosphorylation, compared to MPs from epithelial-like cell lines (OVCAR3 and MCF7). The MPs from mesenchymal-like cells contained increased angiogenic molecules including PDGF, IL8 and angiogenin. The endothelial activation was associated to increased Arf6 expression and MPs secretion. Endothelial activation functionalized an MP dependent pro-tumoral vascular niche promoting cancer cells proliferation, invasiveness, stem cell phenotype and chemoresistance. MPs from cancer and endothelial cells displayed phenotypic heterogeneity, and participated to a functional cross-talk where endothelial activation by cancer MPs resulted in increased secretion of EC-MPs sustaining tumor cells. Such cross-talk may play a role in perfusion independent role of the endothelium.

No MeSH data available.


Related in: MedlinePlus