Ozdian Tomas, Holub Dusan, Maceckova Zuzana, Varanasi Lakshman, Rylova Gabriela, Rehulka Jiri, Vaclavkova Jana, Slavik Hanus, Moudry Pavel, Znojek Pawel, Stankova Jarmila, de Sanctis Juan Bautista, Hajduch Marian, Dzubak Petr
Laboratory of Experimental Medicine, Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacky University and University Hospital in Olomouc, Olomouc, Czech Republic.
Laboratory of Genome Integrity, Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Republic.
J Proteomics. 2017 Jun 6;162:73-85. doi: 10.1016/j.jprot.2017.05.005. Epub 2017 May 3.
Oxaliplatin is widely used to treat colorectal cancer in both palliative and adjuvant settings. It is also being tested for use in treating hematological, esophageal, biliary tract, pancreatic, gastric, and hepatocellular cancers. Despite its routine clinical use, little is known about the responses it induces in cancer cells. Therefore the whole-cell proteomics study was conducted to characterize the cellular response induced by oxaliplatin. Chemosensitive CCRF-CEM cells were treated with oxaliplatin at 29.3μM (5×IC) for 240min (half-time to caspase activation). The proteomes of un-/treated cells were then compared by high-resolution mass spectrometry, revealing 4049 proteins expressed over 3 biological replicates. Among these proteins, 76 were significantly downregulated and 31 significantly upregulated in at least two replicates. In agreement with the DNA-damaging effects of platinum drugs, proteins involved in DNA damage responses were present in both the upregulated and downregulated groups. The downregulated proteins were divided into three subgroups; i) centrosomal proteins, ii) RNA processing and iii) ribosomal proteins, which indicates nucleolar and ribosomal stress. In conclusion, our data supported by further validation experiments indicate the initial cellular response to oxaliplatin is the activation of DNA damage response, which in turn or in parallel triggers nucleolar and ribosomal stress.
We have performed a whole-cell proteomic study of cellular response to oxaliplatin treatment, which is the drug predominantly used in the treatment of colorectal cancer. Compared to its predecessors, cisplatin and carboplatin, there is only a small fraction of studies dedicated to oxaliplatin. From those studies, most of them are focused on modification of treatment regimens or study of oxaliplatin in new cancer diagnoses. Cellular response hasn't been studied deeply and to our best knowledge, this is the first whole-cell proteomics study focused exclusively to this important topic, which can help to understand molecular mechanisms of action.
奥沙利铂广泛用于晚期和辅助治疗结直肠癌。它也正在进行治疗血液学、食管癌、胆管癌、胰腺癌、胃癌和肝细胞癌的试验。尽管它在临床中常规使用，但对其在癌细胞中诱导的反应了解甚少。因此，进行了全细胞蛋白质组学研究以表征奥沙利铂诱导的细胞反应。用29.3μM（5×IC）的奥沙利铂处理化学敏感的CCRF-CEM细胞240分钟（半胱天冬酶激活的半衰期）。然后通过高分辨率质谱比较未处理/处理细胞的蛋白质组，在3个生物学重复中鉴定出4049种表达的蛋白质。在这些蛋白质中，至少在两个重复中有76种显著下调，31种显著上调。与铂类药物的DNA损伤作用一致，参与DNA损伤反应的蛋白质同时存在于上调和下调组中。下调的蛋白质分为三个亚组；i）中心体蛋白，ii）RNA加工和iii）核糖体蛋白，这表明核仁应激和核糖体应激。总之，我们的数据得到进一步验证实验的支持，表明对奥沙利铂的初始细胞反应是DNA损伤反应的激活，这反过来或同时触发核仁应激和核糖体应激。
我们对奥沙利铂治疗的细胞反应进行了全细胞蛋白质组学研究，奥沙利铂是主要用于治疗结直肠癌的药物。与其前身顺铂和卡铂相比，专门针对奥沙利铂的研究只有一小部分。在这些研究中，大多数都集中在治疗方案的调整或奥沙利铂在新癌症诊断中的研究。细胞反应尚未深入研究，据我们所知，这是第一项专门针对这一重要课题的全细胞蛋白质组学研究，有助于理解其分子作用机制。
