Podoplanin increases the migration of human fibroblasts and affects the endothelial cell network formation: A possible role for cancer-associated fibroblasts in breast cancer progression

Abstract

In our previous studies we showed that in breast cancer podoplanin-positive cancer-associated fibroblasts correlated positively with tumor size, grade of malignancy, lymph node metastasis, lymphovascular invasion and poor patients’ outcome. Therefore, the present study was undertaken to assess if podoplanin expressed by fibroblasts can affect malignancy-associated properties of breast cancer cells. Human fibroblastic cell lines (MSU1.1 and Hs 578Bst) overexpressing podoplanin and control fibroblasts were co-cultured with breast cancer MDA-MB-231 and MCF7 cells and the impact of podoplanin expressed by fibroblasts on migration and invasiveness of breast cancer cells were studied in vitro. Migratory and invasive properties of breast cancer cells were not affected by the presence of podoplanin on the surface of fibroblasts. However, ectopic expression of podoplanin highly increases the migration of MSU1.1 and Hs 578Bst fibroblasts. The present study also revealed for the first time, that podoplanin expression affects the formation of pseudo tubes by endothelial cells. When human HSkMEC cells were co-cultured with podoplanin-rich fibroblasts the endothelial cell capillary-like network was characterized by significantly lower numbers of nodes and meshes than in co-cultures of endothelial cells with podoplanin-negative fibroblasts. The question remains as to how our experimental data can be correlated with previous clinical data showing an association between the presence of podoplanin-positive cancer-associated fibroblasts and progression of breast cancer. Therefore, we propose that expression of podoplanin by fibroblasts facilitates their movement into the tumor stroma, which creates a favorable microenvironment for tumor progression by increasing the number of cancer-associated fibroblasts, which produce numerous factors affecting proliferation, survival and invasion of cancer cells. In accordance with this, the present study revealed for the first time, that such podoplanin-mediated effects can affect tube formation by endothelial cells and participate in their pathological properties in the tumor context. Our experimental data were supported by clinical studies. First, when IDC and DCIS were analyzed by immunohistochemistry according to the presence of podoplanin-expressing cells, the numbers of cancer-associated fibroblasts with high expression of this glycoprotein were significantly higher in IDC than in DCIS cases. Second, using immunofluorescence, the co-localization of PDPN-positive CAFs with blood vessels stained with antibody directed against CD34 was observed in tumor stroma of IDC samples.

Dynamic Interstitial Cell Response during Myocardial Infarction Predicts Resilience to Rupture in Genetically Diverse Mice

“…MT1-2 + was mostly limited to CD45 + and sub-epicardial cells at d5. Consistently, SPRING analysis showed that the IR cluster evolved from high expression of Mt1-2 and stromal and fibroblast genes (Gsn [Witke et al, 1995], Pdpn [Suchanski et al, 2017], Angptl4) on d1 to low expression of Mt1-2 accompanied by Myofb markers (Fn1, Acta2, Cthrc1) on d3 ( Figure 4F), suggesting that IR cells rapidly transition to Myofb, in contrast to a previous report suggesting the appearance of Myofb not earlier than d7 post-MI (Farbehi et al, 2019).…”

Sulfatide decreases the resistance to stress-induced apoptosis and increases P-selectin-mediated adhesion: a two-edged sword in breast cancer progression

“…For generation of the GAL3ST1-expressing vector, the human GAL3ST1 cDNA was amplified by PCR from the MCF7 cDNA library using the primers presented in Additional file 1 : Table S1. The resulting insert was cloned into a pRRL-CMV-IRES-PURO vector as described previously [ 18 ] and named here as pRRL-CMV-GAL3ST1-IRES-PURO. For lentivirus production, packaging LentiX 293 T cells were co-transfected at 50–60% confluence with 20 μg of expression or control vector, 10 μg pMDL-g/p-RRE, 5 μg pRSV-REV and 5 μg pMk-VSVG (kindly provided by Dr D. Trono, École Polytechnique Fédérale de Lausanne, Switzerland) using polyethylenimine (Sigma-Aldrich).…”

Fibroblastic Reticular Cell Response to Dendritic Cells Requires Coordinated Activity of Podoplanin, CD44 and CD9

“…In mesenchymal stromal cells, it has been shown that podoplanin expression is required for Rac1-dependent migration (30). Furthermore, podoplanin drives cell migration of CAFs (31) and cancer cells (32,33), and as such plays a role in several stages of the metastasis process (1). Podoplanin is expressed at the invasive front of tumours (34), and more specifically recruited to the adhesion ring of invadopodia where it localizes in lipid rafts (35).…”