Proteomic Profiling of Paclitaxel Treated Cells Identifies a Novel Mechanism of Drug Resistance Mediated by PDCD4.
Proteomic Profiling of Paclitaxel Treated Cells Identifies a Novel Mechanism of Drug Resistance Mediated by PDCD4.
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DOI：
10.1021/acs.jproteome.5b00004
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发表时间：
2015-05
影响因子：
4.4
通讯作者：
Hui Xu;Noah E. Dephoure;Huiying Sun;Haiyuan Zhang;F. Fan;Jiawei Liu;Xuelian Ning;S. Dai;
中科院分区：
文献类型：
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作者：
Hui Xu;Noah E. Dephoure;Huiying Sun;Haiyuan Zhang;F. Fan;Jiawei Liu;Xuelian Ning;S. Dai;
紫杉醇（PTX）是一种广泛使用的化疗药物，对许多癌症有效。为了阐明PTX靶向的细胞途径并鉴定PTX抗性的新机制，我们使用基于SILAC的定量蛋白质组学方法来评估HeLa细胞中细胞蛋白质丰度的总体变化。我们确定了347个蛋白质参与了一些生物过程，包括纺锤体组装，有丝分裂退出，和细胞外粘附，其丰度的变化后，PTX治疗。值得注意的是，参与翻译抑制的肿瘤抑制因子PDCD 4被PTX下调。我们证明PDCD 4是一种细胞周期调节蛋白，其丰度的变化足以改变多种人类癌细胞系对PTX的敏感性。PDCD 4-RNA复合物的免疫沉淀和RT-PCR显示，PDCD 4介导的PTX敏感性通过其与UBE 2S的mRNA相互作用而起作用，UBE 2S是一种对有丝分裂退出至关重要的泛素K11连接缀合酶。最后，肺癌组织中高水平的PDCD 4与接受PTX辅助治疗的受检肺癌患者的较长总生存时间呈正相关。因此，我们对紫杉醇靶点的蛋白质组学筛选不仅通过PDCD 4-有丝分裂退出调节轴提供了对紫杉醇细胞耐药性的新见解，而且还为基于紫杉醇的个性化化疗治疗肺癌提供了预测性生物标志物。
Paclitaxel (PTX) is a widely used chemotherapeutic drug effective against numerous cancers. To elucidate cellular pathways targeted by PTX and identify novel mechanisms of PTX resistance, we used a SILAC based quantitative proteomic approach to evaluate global changes of cellular protein abundance in HeLa cells. We identified 347 proteins involved in a number of biological processes including spindle assembly, mitotic exit, and extracellular adhesion whose abundance changes upon PTX treatment. Notably, the tumor suppressor PDCD4 involved in translation suppression was down-regulated by PTX. We demonstrated that PDCD4 is a cell-cycle regulated protein and that changes in its abundance are sufficient to alter PTX sensitivity in multiple human cancer cell lines. Immunoprecipitation of PDCD4-RNA complexes and RT-PCR revealed that PDCD4 mediated PTX sensitivity acts through its interaction with mRNA of UBE2S, a ubiquitin K11 linkage conjugating enzyme critical for mitotic exit. Lastly, high levels of PDCD4 in lung cancer tissues are positively correlated with the longer overall survival time of the examined lung cancer patients with PTX involved adjuvant therapy. Therefore, our proteomic screen for paclitaxel targets not only provided novel insight into the cellular resistance to paclitaxel via the PDCD4-mitotic exit regulation axis, but also offered a predictive biomarker for paclitaxel-based personalized chemotherapy in the treatment of lung cancer.