NOTCH1 signaling promotes chemoresistance via regulating ABCC1 expression in prostate cancer stem cells
NOTCH1 signaling promotes chemoresistance via regulating ABCC1 expression in prostate cancer stem cells
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NOTCH1信号通过调节前列腺癌干细胞中的ABCC1表达促进化疗耐药
DOI：
10.1007/s11010-014-2069-4
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发表时间：
2014-08-01
影响因子：
4.3
通讯作者：
Xu, Kewei
中科院分区：
文献类型：
--
作者：
Liu, Cheng;Li, Zhuohang;Xu, Kewei
化疗是晚期前列腺癌患者的一种策略，特别是那些去势抵抗性前列腺癌患者。前列腺癌干细胞（PCSC）被认为是治疗干预（包括化疗）后癌症复发的起源。PCSC的化学抗性的潜在机制仍然知之甚少。在本研究中，使用荧光激活细胞分选从LNCaP和PC 3细胞系中分离PCSCs。使用3-（4，5-二甲基-2-噻唑基）-2，5-二苯基-2H-四唑溴化物来测量细胞活力。实时定量PCR和蛋白质印迹用于评估mRNA和蛋白质水平。采用shRNA敲低靶基因表达。染色质免疫沉淀（ChIP）进行探讨ABCC 1表达的详细机制。我们的研究结果表明，分选的PCSC比匹配的非PCSC显示出增强的化学抗性能力。PCSC中激活形式的NOTCH 1（ICN 1）蛋白水平显著较高。用shRNA抑制NOTCH 1可降低PCSC中ABCC 1的表达，并提高PCSC的化疗敏感性。ChIP-PCR结果显示ICN 1可与ABCC 1的启动子区直接结合。结论：NOTCH 1信号转导通路能够反式激活ABCC 1，从而增强PCSCs的耐药能力，这可能是PCSCs耐药的重要机制之一。
Chemotherapy is a strategy for patients with advanced prostate cancer, especially those with castration-resistant prostate cancer. Prostate cancer stem cells (PCSCs) are believed to be the origin of cancer recurrence following therapy intervention, including chemotherapy. The mechanisms underlying the chemoresistance of PCSCs are still poorly understood. In the present study, fluorescence-activated cell sorting was used to isolate PCSCs from LNCaP and PC3 cell lines. 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide was used to measure the cell viability. Quantitative real-time PCR and western blotting were utilized to evaluate the mRNA and protein levels. ShRNA was employed to knock down target gene expression. Chromatin immunoprecipitation (ChIP) was performed to explore the detailed mechanism underlying ABCC1 expression. Our results revealed that the sorted PCSCs showed enhanced chemoresistance ability than matched non-PCSCs. Protein level of activated form of NOTCH1(ICN1) was significantly higher in PCSCs. Inhibition of NOTCH1 with shRNA could decrease ABCC1 expression, and improve chemosensitivity in PCSCs. Finally, ChIP-PCR showed ICN1 could directly bind to the promoter region of ABCC1. In conclusion, NOTCH1 signaling could transactivate ABCC1, resulting in higher chemoresistance ability of PCSCs, which might be one of the important mechanisms underlying the chemoresistance of PCSCs.