About Enhancer

Enhancer ID: E_01_281
Enhancer symbol: --
Species: Human
Position : chr4:77511788-77605824
Biosample name: LNCaP,CWR22Rv1
Experiment class : Low+High throughput
Enhancer type: Enhancer
Disease: Prostate Cancer
DO: DOID:10283
Mesh: D011471
Distance from TSS: Intron
Pubmed ID:  28881808
Enhancer experiment: ChIP-seq,Dual-Luciferase Reporter Assay,ChIP,PCR
Enhancer experiment description: By analyzing AR ChIP-seq data in LNCaP cells, we found that there are two obvious AR binding peaks located within CXCL13 intron I.Corresponding to these two peaks of CHIP-seq results, there were two AR-binding sites (called as ARBS-1 and ARBS-2) in CXCL13 intron I. To investigate the effects of these two ARBS in androgen responsiveness, we constructed two luciferase reporter plasmids (named CXCL13-Luc1 and CXCL13-Luc2) separately containing the two ARBS sequences and then transfected the plasmids into LNCaP cells for luciferase assay. The results of dual-luciferase assay in LNCaP cells showed that ARBS-1 (CXCL13-Luc1) was found to have approximately 3.3-folds increased in luciferase activity by comparing with pGL3-promoter (empty vector), while no obvious luciferase activity changes were observed in ARBS-2 (CXCL13-Luc2; Figure 4D). Furthermore, we generated luciferase reporter plasmids containing key nucleotides mutations within the four ARE sequences of ARBS-1 (called as ARE1-mut, ARE2-mut, ARE3-mut and ARE4-mut).Upon transfection of reporter plasmids into LNCaP cells, the luciferase activities of ARE3-mut and ARE4-mut were obviously decreased by comparing with the luciferase activity of ARBS-1 (CXCL13-Luc1). Moreover, mutant ARE3-mut (mutated at the ARE3 sequence) almost completely abrogated the increased luciferase activity of ARBS-1 (CXCL13-Luc1; Figure 4E). To further verify the ARBS-1 of CXCL13 gene is the androgen responsive element (ARE), CHIP assays were carried out next.Shown as in Figure 4G, PCR analysis using specific primers designed to amplify the ARBS-1 region revealed a significant increment of AR binding in response to Mib stimulation, suggesting this region is the bona fide AR-binding region within CXCL13 enhancer.

About Target gene

Target gene : CXCL13(ANGIE,ANGIE2,BCA-1,BCA1,BLC,BLR1L,SCYB13)
Strong evidence: --
Less strong evidence: ChIP-seq,Dual-Luciferase Reporter Assay,ChIP,PCR
Target gene experiment description: By analyzing AR ChIP-seq data in LNCaP cells, we found that there are two obvious AR binding peaks located within CXCL13 intron I.Corresponding to these two peaks of CHIP-seq results, there were two AR-binding sites (called as ARBS-1 and ARBS-2) in CXCL13 intron I. To investigate the effects of these two ARBS in androgen responsiveness, we constructed two luciferase reporter plasmids (named CXCL13-Luc1 and CXCL13-Luc2) separately containing the two ARBS sequences and then transfected the plasmids into LNCaP cells for luciferase assay. The results of dual-luciferase assay in LNCaP cells showed that ARBS-1 (CXCL13-Luc1) was found to have approximately 3.3-folds increased in luciferase activity by comparing with pGL3-promoter (empty vector), while no obvious luciferase activity changes were observed in ARBS-2 (CXCL13-Luc2; Figure 4D). Furthermore, we generated luciferase reporter plasmids containing key nucleotides mutations within the four ARE sequences of ARBS-1 (called as ARE1-mut, ARE2-mut, ARE3-mut and ARE4-mut).Upon transfection of reporter plasmids into LNCaP cells, the luciferase activities of ARE3-mut and ARE4-mut were obviously decreased by comparing with the luciferase activity of ARBS-1 (CXCL13-Luc1). Moreover, mutant ARE3-mut (mutated at the ARE3 sequence) almost completely abrogated the increased luciferase activity of ARBS-1 (CXCL13-Luc1; Figure 4E). To further verify the ARBS-1 of CXCL13 gene is the androgen responsive element (ARE), CHIP assays were carried out next.Shown as in Figure 4G, PCR analysis using specific primers designed to amplify the ARBS-1 region revealed a significant increment of AR binding in response to Mib stimulation, suggesting this region is the bona fide AR-binding region within CXCL13 enhancer.

About TF

TF name : AR(AIS8,DHTR,HUMARA,HYSP1,KD,NR3C4,SBMA,SMAX1,TFM,AR)
TF experiment: PCR
TF experiment description: AR is known to be the primarily function as a transcription factor by binding the enhancers of its target genes.Shown as in PCR analysis using specific primers designed to amplify the ARBS-1 region revealed a significant increment of AR binding in response to Mib stimulation, suggesting this region is the bona fide AR binding region within CXCL13 enhancer.

About Function

Enhancer function : AR bound the enhancer of CXCL13 gene to up-regulate its expression.
Enhancer function experiment: siRNA,Western blot,PCR
Enhancer function
experiment description:
AR is known to be the primarily function as a transcription factor by binding the enhancers of its target genes.Shown as in PCR analysis using specific primers designed to amplify the ARBS-1 region revealed a significant increment of AR binding in response to Mib stimulation, suggesting this region is the bona fide ARbinding region within CXCL13 enhancer.CXCL13 is an AR target gene and involved in AR-mediated cell migration and invasion in primary PCa.

About SNP

SNP ID: --
SNP position: --
SNP experiment: --

Enhancer associated network

The number on yellow line represents the distance between enhancer and target gene

Expression of target genes for the enhancer


Enhancer associated SNPs