Induction of vimentin synthesis in a murine myeloma cell line by TPA is strongly dependent on the composition of the cell culture medium.
MPC-11 mouse plasmacytoma cells virtually lacking intermediate filament (IF) proteins can be induced to synthesize and accumulate the IF protein vimentin by treatment with the tumor promoting phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA). Like MPC-11 cells, X63-Ag8.6.5.3 mouse myeloma cells (Ag8) proved to be vimentin-negative, as assayed by immunoblotting of whole cellular protein using goat antiserum to vimentin and [125I]protein A. Vimentin synthesis could be elicited by a TPA concentration as low as 10(-9) M in cells grown in HB-102 serum-free medium. Transfer of these cells to medium containing 15% fetal calf cerum (FCS) greatly reduced the ability of these cells to synthesize vimentin upon TPA treatment. After 50 generations of culture in the presence of FCS, induction of vimentin synthesis was barely detectable even at a TPA concentration of 10(-6) M. Addition of FCS to cells grown in serum-free medium partially suppressed vimentin induction by TPA. This suppression seems to be due, at least in part, to nondialyzable, heat-sensitive components of FCS, since the dialyzable fraction even enhanced vimentin induction by TPA. When cells grown in the presence of FCS were transferred back to serum-free medium, their ability to synthesize vimentin in response to TPA treatment was readily restored. The individual components of serum-free medium which proved to support vimentin induction by TPA were insulin and the unsaturated fatty acids oleic acid and linoleic acid. An even stronger TPA response could be elicited by a combination of these components.(ABSTRACT TRUNCATED AT 250 WORDS)
Giese
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Max-Planck-Institut für Zellbiologie, Ladenburg bei Heidelberg, Bundesrepublik Deutschland.
Traub
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D000818:Animals; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D003470:Culture Media / Q000032:analysis / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D007381:Intermediate Filament Proteins / Q000032:analysis; D051379:Mice; D009374:Neoplasms, Experimental / Q000032:analysis / Q000378:metabolism* / Q000473:pathology; D010954:Plasmacytoma / Q000032:analysis / Q000378:metabolism* / Q000473:pathology; D011061:Poly A / Q000235:genetics / Q000302:isolation & purification; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D014407:Tumor Cells, Cultured; D014746:Vimentin / Q000096:biosynthesis*
D003470:Culture Media; D007381:Intermediate Filament Proteins; D012333:RNA, Messenger; D014746:Vimentin; D011061:Poly A; D013755:Tetradecanoylphorbol Acetate