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

• Published in: PloS one 2017-09, Vol.12 (9), p.e0184970-e0184970
• Main Authors: Suchanski, Jaroslaw, Tejchman, Anna, Zacharski, Maciej, Piotrowska, Aleksandra, Grzegrzolka, Jedrzej, Chodaczek, Grzegorz, Nowinska, Katarzyna, Rys, Janusz, Dziegiel, Piotr, Kieda, Claudine, Ugorski, Maciej
• Format: Article
• Language: English
• Subjects: Adult; Aged; Aged, 80 and over; Biochemistry; Biology and Life Sciences; Biophysics; Blood platelets; Blood vessels; Breast cancer; Breast Neoplasms - metabolism; Breast Neoplasms - pathology; Cancer; Cancer-Associated Fibroblasts - metabolism; Cancer-Associated Fibroblasts - pathology; Carcinoma, Ductal, Breast - metabolism; Carcinoma, Ductal, Breast - pathology; CD34 antigen; Cell adhesion & migration; Cell Line; Cell migration; Cell Movement - physiology; Cell proliferation; Cell survival; Coculture Techniques; Cytokines; Disease Progression; Ectopic expression; Embryology; Endothelial cells; Endothelial Cells - metabolism; Endothelial Cells - pathology; Experimental data; Female; Fibroblasts; Gene expression; Gene Expression Regulation, Neoplastic; Genetic aspects; Glycoproteins; Histology; Humans; Immunofluorescence; Immunohistochemistry; Immunology; Invasiveness; Kinases; Laboratories; Life Sciences; Localization; Lymph nodes; Malignancy; Medicine and Health Sciences; Membrane Glycoproteins - genetics; Membrane Glycoproteins - metabolism; Metastases; Middle Aged; Molecular biology; Motility; Neoplasm Invasiveness - physiopathology; Network formation; Pancreatic cancer; Properties (attributes); Proteins; Research and Analysis Methods; RNA, Messenger - metabolism; Stroma; Tubes; Tumor cell lines; Veterinary medicine
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0184970

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 c