Figure 6.

Effect of 1,25(OH)₂D3 on gelatinolytic activities of MMP 2 and 9 in cultured human uterine fibroid cells. Both immortalized human uterine fibroid (HuLM) cells (A) and primary uterine fibroid cells (B) were serum starved and treated with increasing concentrations of 1,25(OH)₂D3 (0, 1, 10, 100 M and 1000 nM) for 48 h. Each supernatant medium (40 µl/each sample) was analyzed in 7.5% SDS PAGE gelatin zymography as described in the section Materials and methods. The digestion of gelatin in the gel by MMP-2 and MMP-9 enzymes cause the negative (unstained) bands. The pro-MMP-2 (72 kDa) and active MMP-2 (65 kDa) digested bands were detected in the gels which were decreased by 1,25(OH)₂D3. Similarly, the pro-MMP-9 (92 kDa) was also detected in supernatant media in both cell types, which were also decreased by 1,25(OH)₂D3. The active form of MMP-9 was undetectable in supernatant media of both cell types. This experiment was repeated twice with similar results.