Induction of collagenase secretion in human fibroblast cultures by growth promoting factors.

C C Chua,D E Geiman,G H Keller,R L Ladda

doi:10.1016/s0021-9258(18)89004-4

C C Chua, D E Geiman + Show 2 more

Open Access

https://doi.org/10.1016/s0021-9258(18)89004-4

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Abstract

In human foreskin fibroblast cultures, two proteins with Mr 60,000 and 55,000 were found to be induced about 3.5-fold by epidermal growth factor (EGF), platelet-derived growth factor, and beta-transforming growth factor. The induced proteins were identified as procollagenases by immunoprecipitation of induced medium with antibodies to purified human fibroblast collagenase. Collagenase enzyme activity in the medium from EGF-treated cultures was also induced at least 3-fold compared to control cultures. Induction of collagenase was dependent upon de novo protein and RNA synthesis and was observed in the medium 10 h after addition of EGF. Although these growth-promoting factors interact with separate membrane receptors, each induced the secretion of a common protein, suggesting that collagenase may be important in some aspect of mitogenesis, cell mobilization, and migration.

Highlights

Labeling of H F Cells-Human fibroblast cultures were established from neonatal foreskins
We have clearly demonstrated that mitogenic agents induce HF cells to secrete collagenase
Separate membrane receptors, and each has a special role in making cells competent to enter mitosis and subsequently capable to progress through mitosis, leading us to speculate that thecommon response may baen important part of some aspect of mitogenesis (24-26)

Summary

Animal cells in culture are known to secrete both growth

Antibody-EGF-induced and uninduced cultured medium were im-. These secretory proteins are probably important regulatofrs munoprecipitated with 40 pl of 5-fold diluted collagenase antibody various aspectsof cellular growth. A hundred pl of protein A-Sepharosewas added and incubated a t room temperature for 1 h.Immunoprecipitates were collectedby centrifugationand washed six timeswith a solution containing 0.16 M NaC1,0.5% NonidetP-40, 0.03%bovine serum foundthatthepotent mitogenic agents,epidermal growth albumin, and0.1% SDS. Assayswere performed in duplicatbey incubating the activated medium with collagen fibrils in 100-p1reaction mixtures. After 24-h mal growth factor; PDGF, platelet-derivedgrowth factor; 8-TGF, p- incubation at room temperature, reaction mixtures were centrifuged transforming growth factor; FBS, fetal bovine serum; SDS-PAGE, a t 12,000 X g for 10 min a t 4 ‘ C , and an 80-p1 aliquot was counted.

RESULTS

Induction ofcollagenasesecretion in HF cells by EGF

DISCUSSION