Tumor promoter and fibronectin induce actin stress fibers and focal adhesion sites in spreading human erythroleukemia (HEL) cells.
The effects of plasma fibronectin (pFn) and the tumor promoter 12-0-tetradecanoyl-phorbol-13-acetate (TPA) on adhesion and cytoskeletal organization of human erythroleukemia (HEL) cells were studied. HEL cells, that normally grow in suspension, attached rapidly on pFn-coated growth substratum and some cells showed spreading. Upon exposure to TPA most of the cells adhered and showed some degree of spreading also when plated on plastic. The spread cells showed mostly peripheral accumulations of F-actin in addition to actin fibers seen in some of the cells. When the cells were plated in the presence of TPA on pFn or on pFn-fragments, containing the cell binding site, all the cells adhered rapidly, spread extensively, organized prominent F-actin stress fibers and typical ventral plaques of vinculin and alpha-actinin. Both proteins were revealed also in the suspended cells by Western blot analysis. When plated on substratum coated with other pFn-fragments or laminin, the HEL cells did not adhere or spread. Both adhesion on pFn as well as formation of stress fibers in the presence of TPA could be prevented by the synthetic peptide Arg-Gly-Asp-Ser (RGDS). HEL cells were also able to organize typical ventral fibrillar arrays of Fn. Immunostaining and metabolic labeling experiments showed that the cells did not contain or synthesize Fn, indicating that the plaques were formed from exogenous pFn by the cells. The results suggest that Fn and TPA synergistically induce the organization of the actomyosin system in HEL cells by promoting the formation of prominent actin stress fibers and focal adhesion sites.
Järvinen
M
M
Department of Pathology, University of Helsinki, Finland.
Ylänne
J
J
Vartio
T
T
Virtanen
I
I
D000199:Actins / Q000032:analysis; D001345:Autoradiography; D002448:Cell Adhesion / Q000187:drug effects*; D003599:Cytoskeleton / Q000187:drug effects* / Q000648:ultrastructure; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts; D005353:Fibronectins / Q000494:pharmacology*; D005455:Fluorescent Antibody Technique; D006801:Humans; D007118:Immunoassay; D004915:Leukemia, Erythroblastic, Acute / Q000473:pathology*; D010641:Phenotype; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
D000199:Actins; D005353:Fibronectins; D013755:Tetradecanoylphorbol Acetate