ANALYSIS FOR THE MODULATION OF MATRIX METALLOPROTEINASE-9 EXPRESSION IN APOPTOSIS

1. 分化抑制因子(DRF)-1候选因子（Dc）在基质金属蛋白酶(MMP)-9启动子中以α或β的形式存在，与c-Jun分子相似。2. DC基因转染的293T细胞增加了kre - m9结合蛋白的数量。3. 转染Dcβ可下调培养的人角质形成细胞（Kc）的MMP-9分泌。4. 在Kc中，用于分化的高Ca^<2+>浓度（hCa）诱导MMP-9表达和Dc转运到细胞质中。在低Ca^<2+>后的去分化Kc中，DC向细胞核内移动，MMP-9下调。到目前为止，hCa还没有检测到细胞核内的磷酸化c- jun、DNA断裂、TUNEL阳性染色以及可溶性Fas配体浓度的变化。5. 细胞因子TGF-β、TNF-α或IL-1α的加入或紫外线B （UVB）照射均上调Kc中MMP-9的表达。通过TNF-α、IL-1α或UVB检测核中Phospho-c-Jun。荧光素酶测定表明，KRE-M9不仅能被hCa诱导，还能被TNF-α或IL-1α诱导。UVB观察到活Kc数量减少，膜联蛋白V阳性Kc。6. 通过RT-PCR， APAF-1未被上述的细胞因子检测到。7. 在间充质成纤维细胞和HT-1080细胞中也观察到DRF-1。TGF-β或TNF-α也能诱导成纤维细胞中MMP-9的表达。8. 在鳞状细胞癌的病例中，原位酶谱检测不仅在侵袭性病变中检测到明胶溶解活性，而且在角化异常病变中也检测到明胶溶解活性，这种活性被MMP抑制剂1,10-菲罗啉抑制。综上所述，HMP-9参与细胞早期凋亡，Dc对MMP-9的表达有抑制作用。

1. The differentiation represssing factor (DRF)-1 candidate (Dc) by one-hybrid method, which binds to KRE-M9 in the matrix metlloproteinase (MMP)-9 promoter was shown to exist as α or β with similarity to c-Jun molecule. 2. The DC gene-transfected 293T cells increased the amounts of KRE-M9-binding protein. 3. The secretion of MMP-9 from human cultured keratinocytes (Kc) was downregulated by the transfection of Dcβ. 4. In Kc, high Ca^<2+> concentration (hCa) for differentiation induced the MMP-9 expression and the trafficking of Dc into cytoplasm. In the dedifferentiated Kc after low Ca^<2+> back, DC was moved into nuclei with MMP-9 down-regulation. So far, phospho-c-Jun in the nuclei, DNA fragmentation, positive TUNEL staining, and the change in the concentrations of soluble Fas ligand have not been detected by hCa. 5. The addition of each cytokine, TGF-β, TNF-α or IL-1α, or the ultraviolet B (UVB) irradiation upregulated MMP-9 expression in Kc. Phospho-c-Jun was detected in the nuclei by TNF-α, IL-1α, or UVB. Luciferase assay showed that KRE-M9 was responsible for MMP-9 induction not only by hCa, but also by TNF-α or IL-1α. By UVB, the reduced number of live Kc, and annexin V positive Kc were observed. 6. By RT-PCR, APAF-1 was not detected with an ycytokine as described above. 7. DRF-1 was also observed in mesenchymal fibroblasts and HT-1080 cells. TGF-β or TNF-α also induced the MMP-9 expression from fibroblasts. 8. In situ zymography in the cases of squamous cell carcinoma detected gelatinolytic activity not only in the invasive lesions, but also in the dyskeratotic lesions, which was inhibited by MMP inhibitor, 1,10-phenanthroline. In conclusion, the involvement of HMP-9 in early apoptosis and the inhibition of MMP-9 expression by Dc were suggested.