The androgen/androgen receptor (AR)-signaling axis plays a central role in prostate cancer (PCa). Upon androgen-binding the AR dimerizes with another AR, and translocates into the nucleus where the AR-dimer activates/inactivates androgen-dependent genes. Consequently, treatments for PCa are commonly based on androgen deprivation therapy (ADT). The clinical benefits of ADT are only transitory and most tumors develop mechanisms allowing the AR to bypass its need for physiological levels of circulating androgens. Clinical failure of ADT is often characterized by the synthesis of a constitutively active AR splice variant, termed AR-V7. AR-V7 mRNA expression is considered as a resistance mechanism following ADT. AR-V7 no longer needs androgenic stimuli for nuclear entry and/or dimerization. Our goal was to mechanistically decipher the interaction between full-length AR (AR-FL) and AR-V7 in AR-null HEK-293 cells using the NanoLuc Binary Technology under androgen stimulation and deprivation conditions. Our data point toward a hypothesis that AR-FL/AR-FL homodimers form in the cytoplasm, whereas AR-V7/AR-V7 homodimers localize in the nucleus. However, after androgen stimulation, all the AR-FL/AR-FL, AR-FL/AR-V7 and AR-V7/AR-V7 dimers were localized in the nucleus. We showed that AR-FL and AR-V7 form heterodimers that localize to the nucleus, whereas AR-V7/AR-V7 dimers were found to localize in the absence of androgens in the nucleus.

中文翻译：

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NanoLuc 二元技术作为一种方法论：研究雄激素受体和雄激素受体剪接变体 V7 同二聚体和异二聚体定位的重要新工具

雄激素/雄激素受体 (AR) 信号轴在前列腺癌 (PCa) 中发挥核心作用。雄激素结合后，AR 与另一个 AR 形成二聚体，并易位到细胞核中，其中 AR 二聚体激活/失活雄激素依赖性基因。因此，PCa 的治疗通常基于雄激素剥夺疗法 (ADT)。ADT 的临床益处只是暂时的，大多数肿瘤都会发展出允许 AR 绕过其对循环雄激素生理水平的需求的机制。ADT 的临床失败通常以合成具有组成型活性的 AR 剪接变体（称为 AR-V7）为特征。AR-V7 mRNA 表达被认为是 ADT 后的耐药机制。AR-V7 不再需要雄激素刺激来进入核和/或二聚化。我们的目标是在雄激素刺激和剥夺条件下使用 NanoLuc 二元技术从机械角度破译 AR 无效 HEK-293 细胞中全长 AR (AR-FL) 和 AR-V7 之间的相互作用。我们的数据表明 AR-FL/AR-FL 同二聚体在细胞质中形成，而 AR-V7/AR-V7 同二聚体位于细胞核中。然而，雄激素刺激后，所有 AR-FL/AR-FL、AR-FL/AR-V7 和 AR-V7/AR-V7 二聚体均定位于细胞核。我们发现 AR-FL 和 AR-V7 形成定位于细胞核的异二聚体，而 AR-V7/AR-V7 二聚体被发现定位于细胞核中不存在雄激素的情况。