阻断雄激素受体信号是晚期前列腺癌（PCa）治疗的主要方法。然而，单一靶向该通路的药物产生的获得性耐药导致了致命的去势耐药性前列腺癌的发展。联合治疗方法代表了治疗晚期疾病的有希望的策略。在这里，我们探索了一种基于shRNA/siRNA能够本质上起到miRNA作用，并通过种子序列互补性同时诱导多个靶点的RNA干扰的PCa治疗策略。我们开发了一个包含所有可能种子序列组合的shRNA文库，以识别当其在PCa细胞中表达时最有效地减少细胞生长和存活能力的种子序列。对其中一些RNAi序列的验证表明，其毒性效应与种子序列与AR核辅助基因和必需基因的3' UTR的互补性相关。事实上，含有已鉴定出的有毒种子序列的siRNA的表达导致了AR介导的基因表达的全局抑制和细胞周期基因的表达降低。在小鼠中的测试中，有毒的shRNA也抑制了去势耐药性前列腺癌，并在既有肿瘤中展现出治疗效果。我们的发现强调了雄激素信号网络的RNA干扰作为一种治疗PCa的有希望的策略。© 2023作者。

Blocking androgen receptor signaling is the mainstay of therapy for advanced prostate cancer (PCa). However, acquired resistance to single agents targeting this pathway results in the development of lethal castration-resistant PCa. Combination therapy approaches represent a promising strategy for the treatment of advanced disease. Here, we explore a therapeutic strategy for PCa based on the ability of shRNAs/siRNAs to function essentially as miRNAs and, via seed sequence complementarity, induce RNA interference of numerous targets simultaneously. We developed a library that contained shRNAs with all possible seed sequence combinations to identify those ones that most potently reduce cell growth and viability when expressed in PCa cells. Validation of some of these RNAi sequences indicated that the toxic effect is associated with seed sequence complementarity to the 3' UTR of AR coregulatory and essential genes. In fact, expression of siRNAs containing the identified toxic seed sequences led to global inhibition of AR-mediated gene expression and reduced expression of cell-cycle genes. When tested in mice, the toxic shRNAs also inhibited castration-resistant PCa and exhibited therapeutic efficacy in pre-established tumors. Our findings highlight RNAi of androgen signaling networks as a promising therapeutic strategy for PCa.© 2023 The Author(s).