Proteomic analysis of cisplatin- and oxaliplatin-induced phosphorylation in proteins bound to Pt-DNA adducts.
Proteomic analysis of cisplatin- and oxaliplatin-induced phosphorylation in proteins bound to Pt-DNA adducts.
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DOI：
10.1039/d0mt00194e
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发表时间：
2020-11
期刊：
影响因子：
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通讯作者：
Xin Yuan;Wanjun Zhang;Yafeng He;Jian Yuan;Dongfan Song;Hao Chen;Weijie Qin;X. Qian;Hanyang Yu;Zijian Guo
中科院分区：
文献类型：
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作者：
Xin Yuan;Wanjun Zhang;Yafeng He;Jian Yuan;Dongfan Song;Hao Chen;Weijie Qin;X. Qian;Hanyang Yu;Zijian Guo
顺铂和奥沙利铂是目前广泛使用的抗肿瘤化疗药物，它们的活性谱不同。这种以铂为基础的药物的疗效被认为至少部分是由于铂-DNA加合物的形成，随后是DNA损伤反应和最终的细胞凋亡。然而，目前尚不清楚顺铂和奥沙利铂引起的这些DNA损伤是否会在细胞信号通路中引起不同的反应。在这里，进行了一项无标记的比较蛋白质组学研究，以描述具有不同配体身份和几何形状的铂-DNA探针的蛋白质磷酸化模式。用LC-MS/MS对识别顺铂和奥沙利铂不同DNA损伤的磷酸化蛋白质进行了浓缩和分析。蛋白质组学分析表明，顺铂主要影响参与mRNA加工的蛋白质，而染色质组织和rRNA加工是奥沙利铂影响的两个主要生物学过程。Western blotting也验证了YBX1和UBF1两种蛋白的位点特异性磷酸化水平的变化。特别是，铂类药物在结肠癌和肝癌细胞系中的作用是下调UBF1的S484磷酸化，UBF1是一种负责核糖体DNA转录激活的关键转录因子，这意味着抑制核糖体的生物发生可能参与了铂类药物的细胞毒性机制。总的来说，这些结果直接反映了顺铂和奥沙利铂引发的不同的蛋白质磷酸化模式，也可能为未来铂类抗肿瘤药物的机制研究提供宝贵的资源。
Cisplatin and oxaliplatin are widely used anti-tumour chemotherapeutic agents with different spectra of activity. The therapeutic efficacy of such platinum-based drug is believed to, at least in part, result from formation of Pt-DNA adducts, followed by DNA damage response and ultimately apoptosis. However, it remains unclear whether these DNA lesions caused by cisplatin and oxaliplatin elicit distinct reactions in cellular signaling pathways. Here, a label-free comparative proteomic study was performed to profile the protein phosphorylation patterns using Pt-DNA probes with different ligand identities and geometries. Phosphorylated proteins recognizing different cisplatin- and oxaliplatin-DNA lesions were enriched and analyzed on LC-MS/MS. Proteomic analysis revealed that cisplatin mainly affected proteins involved in mRNA processing, while chromatin organization and rRNA processing are two major biological processes influenced by oxaliplatin. Changes to site-specific phosphorylation levels of two proteins YBX1 and UBF1 were also validated by Western blotting. In particular, platinum drug treatment in colon and liver cancer cell lines down-regulated S484 phosphorylation of UBF1, which is an essential transcription factor responsible for ribosomal DNA transcription activation, implying that inhibition of ribosome biogenesis might be involved in the cytotoxic mechanism of platinum drugs. Collectively, these results directly reflected distinct protein phosphorylation patterns triggered by cisplatin and oxaliplatin, and could also provide valuable resources for future mechanistic studies of platinum-based anti-tumour agents.