The Zinc Finger Protein RREB-1 is a Co-Regulator of the Androgen Receptor: Implications for the Role of the Ras Pathway in Enhancing Androgenic Signaling in Prostate Cancer

Mukhopadhyay NK, Cinar B, Mukhopadhyay L, Lutchman M, Ferdinand AS, Kim J, Chung LW, Adam RM, Ray SK, Leiter AB, Richie JP, Liu BC, Freeman MR
Source: Mol Endocrinol
Publication Date: (2007)
Issue: 21(9): 2056-2070
Research Area:
Cancer Research/Cell Biology
Cells used in publication:
Species: human
Tissue Origin: prostate
Nucleofector® I/II/2b
Androgen receptor (AR) plays an important role in normal prostate function as well as in the etiology of prostate cancer. Activation of AR is dictated by hormone binding and by interactions with co-regulators. Several of these co-regulators are known targets of Ras related signals. Recent evidence suggests that Ras activation may play a causal role in the progression of prostate cancer toward a more malignant and hormone-insensitive phenotype. In the present study, we used a transcription factor-transcription factor (TF-TF) interaction array method to identify the zinc finger protein, Ras-responsive element binding protein (RREB-1), as a partner and co-regulator of AR. In LNCaP prostate cancer cells, RREB-1 was found to be present in a complex with endogenous AR as determined by co-immunoprecipitation, GST pull down, and immunofluorescence analyses. RREB-1 bound to the prostate specific antigen (PSA) promoter as assessed by chromatin immunoprecipitation. Transient expression of RREB-1 downregulated AR-mediated promoter activity and suppressed expression of PSA protein. The repressor activity of RREB-1 was significantly attenuated by co-transfection of activated Ras. Moreover, expression of the dominant negative N-17- Ras or, alternatively, use of the MEK inhibitor PD98059, abolished the effect of Ras in attenuating RREB-1 mediated repression. Furthermore, inhibition of RREB-1 expression by RNA interference enhanced the effect of Ras on PSA promoter activity and PSA expression. In addition, activation of the Ras pathway depleted AR from the RREB-1/AR complex. Collectively, our data for the first time identify RREB-1 as a repressor of AR and further implicate the Ras/MEK pathway as a likely antagonist of the inhibitory effects of RREB-1 on androgenic signaling.