The androgen receptor (AR) is a key factor that regulates the behavior and fate 
of prostate cancer cells. The AR-regulated network is activated when AR binds 
enhancer elements and modulates specific enhancer-promoter looping. 
Kallikrein-related peptidase 3 (KLK3), which codes for prostate-specific antigen 
(PSA), is a well-known AR-regulated gene and its upstream enhancers produce 
bidirectional enhancer RNAs (eRNAs), termed KLK3e. Here, we demonstrate that 
KLK3e facilitates the spatial interaction of the KLK3 enhancer and the KLK2 
promoter and enhances long-distance KLK2 transcriptional activation. KLK3e 
carries the core enhancer element derived from the androgen response element III 
(ARE III), which is required for the interaction of AR and Mediator 1 (Med1). 
Furthermore, we show that KLK3e processes RNA-dependent enhancer activity 
depending on the integrity of core enhancer elements. The transcription of KLK3e 
was detectable and its expression is significantly correlated with KLK3 (R(2) = 
0.6213, P < 5 × 10(-11)) and KLK2 (R(2) = 0.5893, P < 5 × 10(-10)) in human 
prostate tissues. Interestingly, RNAi silencing of KLK3e resulted in a modest 
negative effect on prostate cancer cell proliferation. Accordingly, we report 
that an androgen-induced eRNA scaffolds the AR-associated protein complex that 
modulates chromosomal architecture and selectively enhances AR-dependent gene 
expression.