Ex [38]. Thus, COUP-TF II probably represses the AR transactivation by a mechanism similar to that for HNF-3a. In contrast, p300, another AR activator, was not able to derepress COUP-TF II-induced suppression of AR transactivation. This is consistent with the fact that p300 activates AR transactivation by purchase Eliglustat inducing the open-structure of chromatin through histone acetylation [47,55], but not by bridging the DBD/LBD complex of AR. This notion is further supported by our results showing that the HDAC inhibitors TSA, NaBut, and NIC were not able to recover the COUP-TF II-induced repression of AR transactivation. AR also performs a crucial function in prostate cancer cell proliferation, and thus the levels of COUP-TF II expression may MNS affect prostate cancer growth. Consistent with this prediction, COUP-TF II expression is down-regulated in prostate cancers as compared with the normal prostate in an animal model of prostate cancer, namely Myc-driven transgenic mice [56]. Further, our data show that COUP-TF II expression in human prostate cancer cell lines is strongly down-regulatedcompared to a normal prostate cell line (Figure 1A). Therefore, COUP-TF II may be associated with the development and progression of prostate cancers, possibly by virtue of its function as an AR corepressor. COUP-TF II has been also reported to inhibit cell growth by blocking cell cycle in MDA-MB-435 cells, ERa-positive and COUP-TF II-negative breast cancer cells [24]. Induction of COUP-TF II in MDA-MB-435 cells resulted in reduced growth, in which cell progression was delayed at G2/M transition phase as a result of the reduction of cdk2 activity. It will be worthwhile to investigate whether cell arrest function of COUP-TF II is also observed in prostate cancer cells and whether the function is related with its inhibitory function of AR transactivation. In the present study, we have shown that COUP-TF II modulates AR function in prostate cancer cells, affecting androgen-dependent cell proliferation. COUP-TF II prevents the N/C terminal interaction of AR, inhibits AR recruitment to its target promoter, and competes with AR coactivators to modulate AR transactivation. The ability of COUP-TF II to repress AR function and inhibit the growth of prostate cancer cells makes COUP-TF II a new candidate as a therapeutic target for prostate cancers.COUP-TF II Inhibits AR TransactivationFigure 6. COUP-TF II inhibits AR recruitment to the PSA 18325633 promoter and competes with AR coactivators to modulate AR transactivation. (A) COUP-TF II inhibits the recruitment of AR to PSA enhancer. LNCaP cells were infected with AdCOUP-TF II or AdGFP. After 24 h of infection, cells were treated with 10 nM DHT or vehicle for 6 h, and then harvested for ChIP assays. Anti-AR antibody (PG-21) was used for immunoprecipitation. Immunoprecipitates were analyzed by PCR using a specific primer pair spanning the AR binding site of the PSA enhancer region. A control PCR for non-specific immunoprecipitation was performed using specific primers to the b-actin coding region. (B) AR coactivators relieve the COUP-TF II-mediated repression of AR transactivation. PPC-1 cells were cotransfected with 50 ng of AR, 250 ng of COUP-TF II and 500 ng of AR coactivator expression plasmids. (C) ARA70 relieves COUP-TF II repression of AR transactivation in a dose-dependent manner. PPC-1 cells were transfected as in “B” with increasing concentration (250, 500, and 1000 ng) of ARA70. (D) COUP-TF II represses ARA70-elevated AR tr.Ex [38]. Thus, COUP-TF II probably represses the AR transactivation by a mechanism similar to that for HNF-3a. In contrast, p300, another AR activator, was not able to derepress COUP-TF II-induced suppression of AR transactivation. This is consistent with the fact that p300 activates AR transactivation by inducing the open-structure of chromatin through histone acetylation [47,55], but not by bridging the DBD/LBD complex of AR. This notion is further supported by our results showing that the HDAC inhibitors TSA, NaBut, and NIC were not able to recover the COUP-TF II-induced repression of AR transactivation. AR also performs a crucial function in prostate cancer cell proliferation, and thus the levels of COUP-TF II expression may affect prostate cancer growth. Consistent with this prediction, COUP-TF II expression is down-regulated in prostate cancers as compared with the normal prostate in an animal model of prostate cancer, namely Myc-driven transgenic mice [56]. Further, our data show that COUP-TF II expression in human prostate cancer cell lines is strongly down-regulatedcompared to a normal prostate cell line (Figure 1A). Therefore, COUP-TF II may be associated with the development and progression of prostate cancers, possibly by virtue of its function as an AR corepressor. COUP-TF II has been also reported to inhibit cell growth by blocking cell cycle in MDA-MB-435 cells, ERa-positive and COUP-TF II-negative breast cancer cells [24]. Induction of COUP-TF II in MDA-MB-435 cells resulted in reduced growth, in which cell progression was delayed at G2/M transition phase as a result of the reduction of cdk2 activity. It will be worthwhile to investigate whether cell arrest function of COUP-TF II is also observed in prostate cancer cells and whether the function is related with its inhibitory function of AR transactivation. In the present study, we have shown that COUP-TF II modulates AR function in prostate cancer cells, affecting androgen-dependent cell proliferation. COUP-TF II prevents the N/C terminal interaction of AR, inhibits AR recruitment to its target promoter, and competes with AR coactivators to modulate AR transactivation. The ability of COUP-TF II to repress AR function and inhibit the growth of prostate cancer cells makes COUP-TF II a new candidate as a therapeutic target for prostate cancers.COUP-TF II Inhibits AR TransactivationFigure 6. COUP-TF II inhibits AR recruitment to the PSA 18325633 promoter and competes with AR coactivators to modulate AR transactivation. (A) COUP-TF II inhibits the recruitment of AR to PSA enhancer. LNCaP cells were infected with AdCOUP-TF II or AdGFP. After 24 h of infection, cells were treated with 10 nM DHT or vehicle for 6 h, and then harvested for ChIP assays. Anti-AR antibody (PG-21) was used for immunoprecipitation. Immunoprecipitates were analyzed by PCR using a specific primer pair spanning the AR binding site of the PSA enhancer region. A control PCR for non-specific immunoprecipitation was performed using specific primers to the b-actin coding region. (B) AR coactivators relieve the COUP-TF II-mediated repression of AR transactivation. PPC-1 cells were cotransfected with 50 ng of AR, 250 ng of COUP-TF II and 500 ng of AR coactivator expression plasmids. (C) ARA70 relieves COUP-TF II repression of AR transactivation in a dose-dependent manner. PPC-1 cells were transfected as in “B” with increasing concentration (250, 500, and 1000 ng) of ARA70. (D) COUP-TF II represses ARA70-elevated AR tr.
