Numb-/low Enriches a Castration-Resistant Prostate Cancer Cell Subpopulation Associated with Enhanced Notch and Hedgehog Signaling
Numb-/low Enriches a Castration-Resistant Prostate Cancer Cell Subpopulation Associated with Enhanced Notch and Hedgehog Signaling
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Numb（-/低）丰富了与增强的 Notch 和 Hedgehog 信号相关的去势抗性前列腺癌细胞亚群
DOI：
10.1158/1078-0432.ccr-17-0913
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发表时间：
2017-11-01
影响因子：
11.5
通讯作者：
Gao, Wei-Qiang
中科院分区：
文献类型：
--
作者：
Guo, Yanjing;Zhang, Kai;Gao, Wei-Qiang
目的：阐明Numb在前列腺癌中的作用和分子机制，以及Numb(-/低)前列腺癌细胞在抗去势中的功能贡献。实验设计：使用多种肿瘤数据集和来自人和小鼠的前列腺癌组织来评估Numb的表达。本实验研究了Numb基因在人前列腺癌细胞系中的过表达和敲除的生物学效应。此外，我们开发了一种可靠的方法来区分内源性Numb蛋白表达高或低的前列腺癌细胞群体，使用基于Numb启动子的慢病毒报告系统。然后测试Numb(-/低)和Numb(高)前列腺癌细胞对雄激素剥夺治疗(ADT)的反应的差异。使用荧光素酶报告分析、免疫印迹和实时定量PCR分析了Numb可能下游因素。结果：我们发现Numb表达下调，并且与前列腺癌进展呈负相关。在功能上，Numb通过抑制Notch和Hedgehog信号，在异种移植前列腺癌的生长和抗去势前列腺癌的发展中发挥了抑制作用。使用基于Numer型启动子慢病毒报告系统，我们能够区分Numb(-/Low)前列腺癌细胞和NumbHigh细胞。Numb(-/Low)前列腺癌细胞较小且静止，优先表达Notch和Hedgehog下游基因和干细胞相关基因，并与更大的ADT耐药性相关。抑制Notch和Hedgehog信号通路显著增加ADT对Numb(-/Low)细胞的凋亡率。结论：Numb(-/Low)丰富了与Notch和Hedgehog信号失控相关的抗去势前列腺癌细胞亚群。(C)2017年AACR。
Purpose: To elucidate the role and molecular mechanism of Numb in prostate cancer and the functional contribution of Numb(-/low) prostate cancer cells in castration resistance.Experimental Design: The expression of Numb was assessed using multiple Oncomine datasets and prostate cancer tissues from both humans and mice. The biological effects of the overexpression and knockdown of Numb in human prostate cancer cell lines were investigated in vitro and in vivo. In addition, we developed a reliable approach to distinguish between prostate cancer cell populations with a high or low endogenous expression of Numb protein using a Numb promoter-based lentiviral reporter system. The difference between Numb(-/low) and Numb(high) prostate cancer cells in the response to androgen-deprivation therapy (ADT) was then tested. The likely downstream factors of Numb were analyzed using luciferase reporter assays, immunoblotting, and quantitative real-time PCR.Results: We show here that Numb was downregulated and negatively correlated with prostate cancer advancement. Functionally, Numb played an inhibitory role in xenograft prostate tumor growth and castration-resistant prostate cancer development by suppressing Notch and Hedgehog signaling. Using a Numb promoter-based lentiviral reporter system, we were able to distinguish Numb(-/low) prostate cancer cells from Numbhigh cells. Numb(-/low) prostate cancer cells were smaller and quiescent, preferentially expressed Notch and Hedgehog downstream and stem-cell-associated genes, and associated with a greater resistance to ADT. The inhibition of the Notch and Hedgehog signaling pathways significantly increased apoptosis in Numb(-/low) cells in response to ADT.Conclusions: Numb(-/low) enriches a castration-resistant prostate cancer cell subpopulation that is associated with unregulated Notch and Hedgehog signaling. (C) 2017 AACR.