Enhancer ID: | E_01_201 | |
Enhancer symbol: | MMP9 Enhancer | |
Species: | Human | |
Position : | chr20:44637014-44637468 |
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Biosample name: | Hepatocellular Carcinoma | |
Experiment class : | Low throughput |
Enhancer type: | Enhancer | |
Disease: | Hepatocellular Carcinoma | |
DO: | DOID:684 | |
Mesh: | D006528 | |
Distance from TSS: | <2KB | |
Pubmed ID: | 25522777 |
Enhancer experiment: | Luciferase Reporter Assay | |
Enhancer experiment description: | The MMP9 promoter contains two AP-1 sites (located at -79 bp and -533 bp), a Sp1 site (located at -560 bp) and an NF-κB site (located at -600 bp). To determine whether regulation of MMP9 is related to these cis-acting regulatory elements for transcription factors, several constructs with deletions or mutations were used and have been described in Materials and methods. Relative Luciferase activities of pGL3-MMP9-WT (containing the potential Enhancer element) and several mutant derivatives of the MMP9 Enhancer region for all motifs were transfected in HCCLM3 cell treated without or with Salin (5-20 µmol/l). Experimental cells were transfected with reporter vectors that included the tandem repeat of AP-1, NF-κB or Sp-1 binding sites. Noteworthy, luciferase activity in the cells with the AP-1 construct was significantly reduced by treatment with Salin at 10-20 µmol/l, whereas luciferase activity containing the NF-κB or Sp-1 binding site construct showed no statistically significant changes in the cells treated with Salin (Fig. 3A). These results showed that both AP-1 sites in the MMP9 promoter were essential for Salin-regulated MMP9 enhancer region activity. |
Target gene : | MMP9(CLG4B,GELB,MANDP2,MMP-9) | |
Strong evidence: | -- | |
Less strong evidence: | Luciferase Reporter Assay | |
Target gene experiment description: | The MMP9 promoter contains two AP-1 sites (located at -79 bp and -533 bp), a Sp1 site (located at -560 bp) and an NF-κB site (located at -600 bp). To determine whether regulation of MMP9 is related to these cis-acting regulatory elements for transcription factors, several constructs with deletions or mutations were used and have been described in Materials and methods. Relative Luciferase activities of pGL3-MMP9-WT (containing the potential Enhancer element) and several mutant derivatives of the MMP9 Enhancer region for all motifs were transfected in HCCLM3 cell treated without or with Salin (5-20 µmol/l). Experimental cells were transfected with reporter vectors that included the tandem repeat of AP-1, NF-κB or Sp-1 binding sites. Noteworthy, luciferase activity in the cells with the AP-1 construct was significantly reduced by treatment with Salin at 10-20 µmol/l, whereas luciferase activity containing the NF-κB or Sp-1 binding site construct showed no statistically significant changes in the cells treated with Salin (Fig. 3A). These results showed that both AP-1 sites in the MMP9 promoter were essential for Salin-regulated MMP9 enhancer region activity. |
TF name : | JUND(AP-1) | |
TF experiment: | Western blot | |
TF experiment description: | The results revealed that the AP-1 site is critical for Salin-regulated MMP9 Enhancer activity. Western blot analysis indicated that the expression of the components of AP-1 (JunB and JunD) was regulated by Salin (5-20 μmol/l). |
Enhancer function : | -- |
Enhancer function experiment: | -- |
Enhancer function experiment description: |
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SNP ID: | -- | |
SNP position: | -- |
SNP experiment: | -- |
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