Characterization and distribution of extracellular matrix components and receptors in GH3B6 prolactin cells

• Published in: Biology of the cell 2000-08, Vol.92 (5), p.351-362
• Main Authors: de Carvalho, Daniela F., Silva, Karina L., de Oliveira, Désio A.F, Villa-Verde, Déa Maria S., Coelho, Hamilton S., Silva, Luiz-Cláudio F., Nasciutti, Luiz E.
• Format: Article
• Language: English
• Subjects: Animals; Cell Adhesion; Cell Size; Collagen - metabolism; extracellular matrix; Extracellular Matrix Proteins - metabolism; Fibronectins - metabolism; Flow Cytometry; Fluorescent Antibody Technique; Gene Expression Regulation - drug effects; GH3B6 cells; glycosaminoglycans; Glycosaminoglycans - metabolism; Hyaluronan Receptors - metabolism; Laminin - metabolism; Pituitary Gland - cytology; Pituitary Gland - drug effects; Pituitary Gland - metabolism; prolactin; Prolactin - metabolism; Rats; Receptors, Cell Surface - metabolism; Receptors, Fibronectin - metabolism; Receptors, Laminin - metabolism; thyroliberin; Thyrotropin-Releasing Hormone - pharmacology; Tumor Cells, Cultured
• ISSN: 0248-4900; 1768-322X
• DOI: 10.1016/S0248-4900(00)01076-5

GH3B6 cells, a rat pituitary tumor cell line, synthesize and secrete large amounts of prolactin (PRL) in vitro. In the present work, we evaluated the capacity of these cells to express extracellular matrix (ECM) components and receptors in vitro. The expression of laminin (LN), fibronectin (FN) and type IV collagen (CIV) was investigated by immunofluorescence assays. In comparison to PRL distribution, where around 50–70 % of the cells contained PRL concentrated in the Golgi region, a variable immunolabeling for the three ECM components could be observed in the majority of GH3B6 cells. Importantly, this pattern was not modified when cells were cultured in the presence of 30 nM thyroliberin (TRH). The expression of the ECM receptors: α5β1 (FN receptor), α6β1 (LN receptor) and CD44 (hyaluronic acid receptor) could be demonstrated by cytofluorometric analysis. Using biochemical procedures, we analyzed the synthesis and secretion of glycosaminoglycans (GAGs). The cells synthesized and secreted mainly heparan sulfate (75 %) with a minor amount of chondroitin sulfate/dermatan sulfate. In an attempt to evaluate the individual contribution of the ECM components to influence cell morphology and PRL distribution in vitro, GH3B6 cells were cultivated separately on LN, FN and CIV substrates. Under all conditions, it was possible to observe an increase of cell adherence to the substrate, accompanied with changes of cellular morphology, characterized by the appearance of cytoplasmatic processes. However, no changes on PRL distribution could be observed. Our results suggest that endocrine tumor cell lines are involved in synthesis of ECM components and receptors.