Enhancer ID: | E_01_211 | |
Enhancer symbol: | ENH3 Enhancer | |
Species: | Human | |
Position : | chr6:45410618-45417584 |
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Biosample name: | B-CPAP,TPC1 | |
Experiment class : | Low throughput |
Enhancer type: | Enhancer | |
Disease: | -- | |
DO: | -- | |
Mesh: | -- | |
Distance from TSS: | >2KB | |
Pubmed ID: | 25769725 |
Enhancer experiment: | Luciferase Reporter Assay,RT-PCR,ChIP | |
Enhancer experiment description: | Next, the DNA fragments corresponding to ENH1, 2, and 3 were cloned into the pGL3-P2 vector downstream of the luciferase reporter gene and their ability to activate the P2 promoter wasassessed by luciferase assay (Fig. 2F). Only ENH3 determined a strong induction of the reporter gene, suggesting a potent activity of this element on the P2 promoter. Noticeably, the activating effect of ENH3 on the P2 promoter was stronger in TPC1 than in BCPAP cells, in line with the higher level of RUNX2 expression detected in TPC1 as compared with BCPAP cells (Fig. 2B; ref. 16).Furthermore, ENH3 maintained the ability of activating the reporter gene when cloned in the inverted orientation and when cloned in the pGL3 vector containing the sP2 promoter.Finally, we have shown by chromatin immunoprecipitation (ChIP) experiments that the binding of RNA Polymerase II (RNA PolII) on the ENH3 was significantly enriched as compared with two downstream RUNX2 exons (exons 5 and 7; Fig.3A and B). |
Target gene : | RUNX2(AML3,CBF-alpha-1,CBFA1,CCD,CCD1,CLCD,OSF-2,OSF2,PEA2aA,PEBP2aA) | |
Strong evidence: | -- | |
Less strong evidence: | Luciferase Reporter Assay,RT-PCR,ChIP | |
Target gene experiment description: | Next, the DNA fragments corresponding to ENH1, 2, and 3 were cloned into the pGL3-P2 vector downstream of the luciferase reporter gene and their ability to activate the P2 promoter wasassessed by luciferase assay (Fig. 2F). Only ENH3 determined a strong induction of the reporter gene, suggesting a potent activity of this element on the P2 promoter. Noticeably, the activating effect of ENH3 on the P2 promoter was stronger in TPC1 than in BCPAP cells, in line with the higher level of RUNX2 expression detected in TPC1 as compared with BCPAP cells (Fig. 2B; ref. 16).Furthermore, ENH3 maintained the ability of activating the reporter gene when cloned in the inverted orientation and when cloned in the pGL3 vector containing the sP2 promoter.Finally, we have shown by chromatin immunoprecipitation (ChIP) experiments that the binding of RNA Polymerase II (RNA PolII) on the ENH3 was significantly enriched as compared with two downstream RUNX2 exons (exons 5 and 7; Fig.3A and B). |
TF name : | JUND(AP-1) | |
TF experiment: | ChIP | |
TF experiment description: | We performed supershift assay using anti-c-JUN antibodies to con-firm that AP1 binds to site 2,ChIP experiments using anti-c-JUN antibodies showed that c-JUN actively binds to ENH3 in a context of structured Chromatin. |
Enhancer function : | Inhibition of ENH3 activity is a generalmechanismof action of HDACi in different types of cancer |
Enhancer function experiment: | Luciferase Reporter Assay |
Enhancer function experiment description: |
Finally, we tested whether the TSA-repressive effect was mediated by repression of ENH3 as we have shown in thyroid cancer cells. In all cell lines tested, the presence of ENH3 down-stream of the sP2 promoter determined the activation of the reporter gene as already observed in TPC1 and BCPAP cells (Fig.7C). Similarly, upon TSA treatment, the ENH3 activity was repressed in all cell lines, suggesting the possibility to extend to other cancer types the model of ENH3 functioning that we proposed in thyroid cancer. |
SNP ID: | -- | |
SNP position: | -- |
SNP experiment: | -- |
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