Actin in B16 melanoma cells of differing metastatic potential. Effects of trypsin and serum.
Actin is present in cells in monomeric and polymeric (filamentous) forms. Filamentous actin is distributed in Triton-soluble (cytosolic) and Triton-insoluble (cytoskeletal core) fractions. We have used the DNase 1 inhibition assay and immunofluorescence to investigate the distribution of actin in monomeric and polymeric forms in cloned B16 murine melanoma cell lines of low and high metastatic capacity. The protease trypsin caused rounding up and detachment of both cell lines within 5 min. This was associated with almost complete depolymerization of cytosolic actin filaments but the Triton-insoluble cytoskeleton was not quantitatively affected by trypsin treatment. There were quantitative differences between the clones in their response to incubation in the presence or absence of 10% serum. The highly metastatic cell line contained 35% more actin when incubated in the presence of 10% serum, almost completely distributed to the Triton-insoluble cytoskeleton, an effect not seen in the low metastatic cells.
D000199:Actins / Q000502:physiology*; D000818:Animals; D001769:Blood; D002460:Cell Line; D002999:Clone Cells; D003470:Culture Media; D003599:Cytoskeleton / Q000648:ultrastructure; D003850:Deoxyribonuclease I; D005455:Fluorescent Antibody Technique; D008545:Melanoma / Q000473:pathology*; D051379:Mice; D008810:Mice, Inbred C57BL; D009362:Neoplasm Metastasis; D014357:Trypsin / Q000494:pharmacology*
D000199:Actins; D003470:Culture Media; D003850:Deoxyribonuclease I; D014357:Trypsin