Limits...
SDF-1alpha concentration dependent modulation of RhoA and Rac1 modifies breast cancer and stromal cells interaction.

Pasquier J, Abu-Kaoud N, Abdesselem H, Madani A, Hoarau-Véchot J, Thawadi HA, Vidal F, Couderc B, Favre G, Rafii A - BMC Cancer (2015)

Bottom Line: This interaction increases migration from primary sites as well as homing at distant sites.Increased migration was obtained at 50 and 100 ng/ml of SDF-1α; however migration was reduced at 200 ng/ml.The adhesion between breast cancer cells and BMHC was significantly increased by SDF-1α treatment at 200 ng/ml and reduced using a blocking monoclonal antibody against CXCR4.

View Article: PubMed Central - PubMed

Affiliation: Stem Cell and Microenvironment Laboratory, Weill Cornell Medical College in Qatar, Education City, Qatar Foundation, Doha, Qatar. jep2026@qatar-med.cornell.edu.

ABSTRACT

Background: The interaction of SDF-1alpha with its receptor CXCR4 plays a role in the occurrence of distant metastasis in many solid tumors. This interaction increases migration from primary sites as well as homing at distant sites.

Methods: Here we investigated how SDF-1α could modulate both migration and adhesion of cancer cells through the modulation of RhoGTPases.

Results: We show that different concentrations of SDF-1α modulate the balance of adhesion and migration in cancer cells. Increased migration was obtained at 50 and 100 ng/ml of SDF-1α; however migration was reduced at 200 ng/ml. The adhesion between breast cancer cells and BMHC was significantly increased by SDF-1α treatment at 200 ng/ml and reduced using a blocking monoclonal antibody against CXCR4. We showed that at low SDF-1α concentration, RhoA was activated and overexpressed, while at high concentration Rac1 was promoting SDF-1α mediating-cell adhesion.

Conclusion: We conclude that SDF-1α concentration modulates migration and adhesion of breast cancer cells, by controlling expression and activation of RhoGTPases.

No MeSH data available.


Related in: MedlinePlus

Intercellular interactions between cancer cells and Bone Marrow Host Cells (BMHC). a Paraffin-embedded immunohistochemistry. Antibody against CD-10 was used. Picture showed a network of BMHC (brown cells) surrounding cancer cell clusters (blue cells). The insert picture showed the metastatic node the tissue micro-array. b Electronic microscopy imaging. MDA-MB-231 and BMHC or MCF7 and BMHC were co-cultured during 48 h and analyzed by electronic microscopy. A pseudopodia of BMHC with two MDA-MB-231 cells were closely interacting with the pseudopodia (left panel, arrows). Very close interaction between the two cellular membranes of MCF7 and BMHC can be observed with formation of tight junction (right panel, arrows). c Co-culture of BMHC and MDA-MB231 in phase microscopy. Cancer cells are growing on BMHC. Scale bar 250 μm. d Confocal imaging of BMHC and eGFP MDA-MB231 co-culture. BMHC were co-cultured with tumor cells for 3 days. Before imaging by confocal microscopy, co-cultures were stained with Alexa Fluor 594 conjugated-wheat germ agglutinin (WGA). Z-X reconstitution shows that cancer cells (green) are growing on BMHC. Scale bar 10 μm. e Adhesion assay testing the specificity of the adhesion between MDA-MB231 cells and BMHC. BMHC were plated up to 60 % confluency, 50,000 eGFP MDA-MB231 were allowed to adhere for 1 h. HBMEC (human bone marrow endothelial cells) or plastic were used as negative control. f Proliferation assay. MDA-MB231 were plated and counted every 2 days in presence or not of BMHC during 6 days. BMHC were able to increase proliferation of MDA-MB231. g Migration in agarose gel assay. MDA-MB231 cells were seeded in the central well. Media only was poured in the left well as negative control and BMHC were seeded in the right well. Cells could be observed during migration through the agarose gel (black part, wall). The picture represents MDA-MB231 cells migration through the agarose wall to the BMHC well at day 4 (bottom picture, arrows) or to the media only (top picture)
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4522077&req=5

Fig1: Intercellular interactions between cancer cells and Bone Marrow Host Cells (BMHC). a Paraffin-embedded immunohistochemistry. Antibody against CD-10 was used. Picture showed a network of BMHC (brown cells) surrounding cancer cell clusters (blue cells). The insert picture showed the metastatic node the tissue micro-array. b Electronic microscopy imaging. MDA-MB-231 and BMHC or MCF7 and BMHC were co-cultured during 48 h and analyzed by electronic microscopy. A pseudopodia of BMHC with two MDA-MB-231 cells were closely interacting with the pseudopodia (left panel, arrows). Very close interaction between the two cellular membranes of MCF7 and BMHC can be observed with formation of tight junction (right panel, arrows). c Co-culture of BMHC and MDA-MB231 in phase microscopy. Cancer cells are growing on BMHC. Scale bar 250 μm. d Confocal imaging of BMHC and eGFP MDA-MB231 co-culture. BMHC were co-cultured with tumor cells for 3 days. Before imaging by confocal microscopy, co-cultures were stained with Alexa Fluor 594 conjugated-wheat germ agglutinin (WGA). Z-X reconstitution shows that cancer cells (green) are growing on BMHC. Scale bar 10 μm. e Adhesion assay testing the specificity of the adhesion between MDA-MB231 cells and BMHC. BMHC were plated up to 60 % confluency, 50,000 eGFP MDA-MB231 were allowed to adhere for 1 h. HBMEC (human bone marrow endothelial cells) or plastic were used as negative control. f Proliferation assay. MDA-MB231 were plated and counted every 2 days in presence or not of BMHC during 6 days. BMHC were able to increase proliferation of MDA-MB231. g Migration in agarose gel assay. MDA-MB231 cells were seeded in the central well. Media only was poured in the left well as negative control and BMHC were seeded in the right well. Cells could be observed during migration through the agarose gel (black part, wall). The picture represents MDA-MB231 cells migration through the agarose wall to the BMHC well at day 4 (bottom picture, arrows) or to the media only (top picture)

Mentions: Tumor stroma is a composed of multiple cell types; we have previously described [33] the infiltration of ovarian cancer tumors by BMHC (CD9+CD10+). Here using paraffin-embedded immunohistochemistry of primary breast cancer specimen we found a network of BMHC (CD9+CD10+) surrounding cancer cell clusters (Fig. 1a). Electron microscopy analysis of co-cultures of BMHC and MDA-MB231 or MCF7 displayed close interactions with formation of tight junctions (Fig. 1b). When the two cell types were seeded at the same time at a ratio of 1/1, breast cancer cells (BCC) attached preferentially on BMHC compare to plastic or matrigel as shown on phase contrast and selected (x-z) sections, obtained from confocal microscopy (Fig. 1c-d). Adhesion of BCC and BMHCs was stronger than spontaneous adhesion to culture plate or to other cell type HBMEC (Human Bone Marrow Endothelial Cells) (Fig. 1e). We then investigated the functional benefit of such interaction. MDA-MB231 co-cultured with BMHC in serum free cytokine free media displayed a proliferative advantage compared to controls (Fig. 1f). Finally, in order to test the ability of BMHC to attract MDA-MB231, we developed an agarose-based migration assay to evaluate the motility of BCC (Fig. 1g). With this method, BMHC secreted factors rather than the components of extracellular matrix surrogates (such as Matrigel) would be responsible for the migration observed. In this set–up MDA-MB231 displayed increased migration toward BMHC compare to control media.Fig. 1


SDF-1alpha concentration dependent modulation of RhoA and Rac1 modifies breast cancer and stromal cells interaction.

Pasquier J, Abu-Kaoud N, Abdesselem H, Madani A, Hoarau-Véchot J, Thawadi HA, Vidal F, Couderc B, Favre G, Rafii A - BMC Cancer (2015)

Intercellular interactions between cancer cells and Bone Marrow Host Cells (BMHC). a Paraffin-embedded immunohistochemistry. Antibody against CD-10 was used. Picture showed a network of BMHC (brown cells) surrounding cancer cell clusters (blue cells). The insert picture showed the metastatic node the tissue micro-array. b Electronic microscopy imaging. MDA-MB-231 and BMHC or MCF7 and BMHC were co-cultured during 48 h and analyzed by electronic microscopy. A pseudopodia of BMHC with two MDA-MB-231 cells were closely interacting with the pseudopodia (left panel, arrows). Very close interaction between the two cellular membranes of MCF7 and BMHC can be observed with formation of tight junction (right panel, arrows). c Co-culture of BMHC and MDA-MB231 in phase microscopy. Cancer cells are growing on BMHC. Scale bar 250 μm. d Confocal imaging of BMHC and eGFP MDA-MB231 co-culture. BMHC were co-cultured with tumor cells for 3 days. Before imaging by confocal microscopy, co-cultures were stained with Alexa Fluor 594 conjugated-wheat germ agglutinin (WGA). Z-X reconstitution shows that cancer cells (green) are growing on BMHC. Scale bar 10 μm. e Adhesion assay testing the specificity of the adhesion between MDA-MB231 cells and BMHC. BMHC were plated up to 60 % confluency, 50,000 eGFP MDA-MB231 were allowed to adhere for 1 h. HBMEC (human bone marrow endothelial cells) or plastic were used as negative control. f Proliferation assay. MDA-MB231 were plated and counted every 2 days in presence or not of BMHC during 6 days. BMHC were able to increase proliferation of MDA-MB231. g Migration in agarose gel assay. MDA-MB231 cells were seeded in the central well. Media only was poured in the left well as negative control and BMHC were seeded in the right well. Cells could be observed during migration through the agarose gel (black part, wall). The picture represents MDA-MB231 cells migration through the agarose wall to the BMHC well at day 4 (bottom picture, arrows) or to the media only (top picture)
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4522077&req=5

Fig1: Intercellular interactions between cancer cells and Bone Marrow Host Cells (BMHC). a Paraffin-embedded immunohistochemistry. Antibody against CD-10 was used. Picture showed a network of BMHC (brown cells) surrounding cancer cell clusters (blue cells). The insert picture showed the metastatic node the tissue micro-array. b Electronic microscopy imaging. MDA-MB-231 and BMHC or MCF7 and BMHC were co-cultured during 48 h and analyzed by electronic microscopy. A pseudopodia of BMHC with two MDA-MB-231 cells were closely interacting with the pseudopodia (left panel, arrows). Very close interaction between the two cellular membranes of MCF7 and BMHC can be observed with formation of tight junction (right panel, arrows). c Co-culture of BMHC and MDA-MB231 in phase microscopy. Cancer cells are growing on BMHC. Scale bar 250 μm. d Confocal imaging of BMHC and eGFP MDA-MB231 co-culture. BMHC were co-cultured with tumor cells for 3 days. Before imaging by confocal microscopy, co-cultures were stained with Alexa Fluor 594 conjugated-wheat germ agglutinin (WGA). Z-X reconstitution shows that cancer cells (green) are growing on BMHC. Scale bar 10 μm. e Adhesion assay testing the specificity of the adhesion between MDA-MB231 cells and BMHC. BMHC were plated up to 60 % confluency, 50,000 eGFP MDA-MB231 were allowed to adhere for 1 h. HBMEC (human bone marrow endothelial cells) or plastic were used as negative control. f Proliferation assay. MDA-MB231 were plated and counted every 2 days in presence or not of BMHC during 6 days. BMHC were able to increase proliferation of MDA-MB231. g Migration in agarose gel assay. MDA-MB231 cells were seeded in the central well. Media only was poured in the left well as negative control and BMHC were seeded in the right well. Cells could be observed during migration through the agarose gel (black part, wall). The picture represents MDA-MB231 cells migration through the agarose wall to the BMHC well at day 4 (bottom picture, arrows) or to the media only (top picture)
Mentions: Tumor stroma is a composed of multiple cell types; we have previously described [33] the infiltration of ovarian cancer tumors by BMHC (CD9+CD10+). Here using paraffin-embedded immunohistochemistry of primary breast cancer specimen we found a network of BMHC (CD9+CD10+) surrounding cancer cell clusters (Fig. 1a). Electron microscopy analysis of co-cultures of BMHC and MDA-MB231 or MCF7 displayed close interactions with formation of tight junctions (Fig. 1b). When the two cell types were seeded at the same time at a ratio of 1/1, breast cancer cells (BCC) attached preferentially on BMHC compare to plastic or matrigel as shown on phase contrast and selected (x-z) sections, obtained from confocal microscopy (Fig. 1c-d). Adhesion of BCC and BMHCs was stronger than spontaneous adhesion to culture plate or to other cell type HBMEC (Human Bone Marrow Endothelial Cells) (Fig. 1e). We then investigated the functional benefit of such interaction. MDA-MB231 co-cultured with BMHC in serum free cytokine free media displayed a proliferative advantage compared to controls (Fig. 1f). Finally, in order to test the ability of BMHC to attract MDA-MB231, we developed an agarose-based migration assay to evaluate the motility of BCC (Fig. 1g). With this method, BMHC secreted factors rather than the components of extracellular matrix surrogates (such as Matrigel) would be responsible for the migration observed. In this set–up MDA-MB231 displayed increased migration toward BMHC compare to control media.Fig. 1

Bottom Line: This interaction increases migration from primary sites as well as homing at distant sites.Increased migration was obtained at 50 and 100 ng/ml of SDF-1α; however migration was reduced at 200 ng/ml.The adhesion between breast cancer cells and BMHC was significantly increased by SDF-1α treatment at 200 ng/ml and reduced using a blocking monoclonal antibody against CXCR4.

View Article: PubMed Central - PubMed

Affiliation: Stem Cell and Microenvironment Laboratory, Weill Cornell Medical College in Qatar, Education City, Qatar Foundation, Doha, Qatar. jep2026@qatar-med.cornell.edu.

ABSTRACT

Background: The interaction of SDF-1alpha with its receptor CXCR4 plays a role in the occurrence of distant metastasis in many solid tumors. This interaction increases migration from primary sites as well as homing at distant sites.

Methods: Here we investigated how SDF-1α could modulate both migration and adhesion of cancer cells through the modulation of RhoGTPases.

Results: We show that different concentrations of SDF-1α modulate the balance of adhesion and migration in cancer cells. Increased migration was obtained at 50 and 100 ng/ml of SDF-1α; however migration was reduced at 200 ng/ml. The adhesion between breast cancer cells and BMHC was significantly increased by SDF-1α treatment at 200 ng/ml and reduced using a blocking monoclonal antibody against CXCR4. We showed that at low SDF-1α concentration, RhoA was activated and overexpressed, while at high concentration Rac1 was promoting SDF-1α mediating-cell adhesion.

Conclusion: We conclude that SDF-1α concentration modulates migration and adhesion of breast cancer cells, by controlling expression and activation of RhoGTPases.

No MeSH data available.


Related in: MedlinePlus