Effects of dehydroepiandrosterone sulphate on the production of collagenase and gelatinolytic metalloproteinase by rabbit uterine cervical cells in primary cultures

Abstract

Cells from pregnant rabbit uterine cervix in primary cultures produced specific collagenase and gelatinolytic metalloproteinase, both in latent forms. The two enzymes were separated by CM-52 cellulose chromatography. Dehydroepiandrosterone 3-sulphate (DHAS) stimulated the production of the latent collagenase by the cells in a dose-dependent manner and the maximal effect, which was about 2-fold of control cultures, was observed when 1 X 10(-6) M DHAS was added to the medium containing 10% (v/v) foetal calf serum (FCS). DHAS also stimulated the production of the gelatinolytic metalloproteinase. The significant stimulative effects were also confirmed when the cells were incubated with 1 X 10(-7) M DHAS in the medium containing 9% (v/v) dextran-coated charcoal treated FCS and 1% (v/v) FCS. The uptake of [3H]DHAS by the cells from pregnant rabbits was 5-10-fold greater than that from nonpregnant rabbits. When confluent cultures were incubated with 6.5 X 10(-8) M [3H]DHAS for 2 d, 3.4% and 0.03% of total radioactivity added were accumulated as [3H]oestradiol-17 beta ([3H]E2) in the medium and cell layer, respectively. Nevertheless, the addition of 1 X 10(-9) M and 1 X 10(-11) M E2 or dehydroepiandrosterone (DHA) to the confluent cultures caused no significant effect on the collagenase production. These results strongly suggest that the stimulative effect of DHAS on the production of collagenase and gelatinolytic metalloproteinase by the rabbit uterine cervical cells was due to unchanged DHAS and not due to E2 or DHA converted from DHAS.