Inhibition of transketolase by oxythiamine altered dynamics of protein signals in pancreatic cancer cells.
Inhibition of transketolase by oxythiamine altered dynamics of protein signals in pancreatic cancer cells.
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DOI：
10.1186/2162-3619-2-18
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发表时间：
2013
影响因子：
10.9
通讯作者：
Xiao GG
中科院分区：
文献类型：
--
作者：
Wang J;Zhang X;Ma D;Lee WP;Xiao J;Zhao Y;Go VL;Wang Q;Yen Y;Recker R;Xiao GG
氧硫胺 (OT) 是一种抗代谢药类似物，可抑制核糖的非氧化合成，并通过在体外和体内引起 G1 期阻滞来诱导细胞凋亡。然而，其分子机制仍不清楚。在本研究中，采用改良SILAC方法（mSILAC）进行定量蛋白质组学分析，以确定代谢抑制对不同剂量（0μM、5μM、50μM和500μM）和时间点（0h、12h和48h）OT处理的MIA PaCa-2癌细胞中蛋白质表达动态变化的影响。经OT处理的MIA PaCa-2细胞中总共鉴定出52个差异蛋白，其中包括14个磷酸化蛋白。根据动态表达模式，这些蛋白质被分为三个簇：直接下调（簇 1，占总蛋白的 37%）、直立“V”形表达模式（簇 2，占总蛋白的 47.8%）和直立“V”形表达模式（簇 3，占总蛋白的 15.2%）。其中，Annexin A1 表达随 OT 处理而显着下调，且呈时间依赖性，而该蛋白的表达未观察到 OT 剂量依赖性变化。通路分析表明，转酮醇酶的抑制导致与细胞凋亡相关的多个细胞信号通路的变化。蛋白质的时间表达模式表明，OT 通过抑制磷酸激酶的表达，以时间依赖性方式改变蛋白质表达的动态，导致癌细胞凋亡。这项研究的结果表明，单一代谢酶活性的干扰改变了多种细胞信号传导途径。
Oxythiamine (OT), an analogue of anti-metabolite, can suppress the nonoxidative synthesis of ribose and induce cell apoptosis by causing a G1 phase arrest in vitro and in vivo. However, the molecular mechanism remains unclear yet. In the present study, a quantitative proteomic analysis using the modified SILAC method (mSILAC) was performed to determine the effect of metabolic inhibition on dynamic changes of protein expression in MIA PaCa-2 cancer cells treated with OT at various doses (0 μM, 5 μM, 50 μM and 500 μM) and time points (0 h, 12 h and 48 h). A total of 52 differential proteins in MIA PaCa-2 cells treated with OT were identified, including 14 phosphorylated proteins. Based on the dynamic expression pattern, these proteins were categorized in three clusters, straight down-regulation (cluster 1, 37% of total proteins), upright “V” shape expression pattern (cluster 2, 47.8% total), and downright “V” shape pattern (cluster 3, 15.2% total). Among them, Annexin A1 expression was significantly down-regulated by OT treatment in time-dependent manner, while no change of this protein was observed in OT dose-dependent fashion. Pathway analysis suggested that inhibition of transketolase resulted in changes of multiple cellular signaling pathways associated with cell apoptosis. The temporal expression patterns of proteins revealed that OT altered dynamics of protein expression in time-dependent fashion by suppressing phosphor kinase expression, resulting in cancer cell apoptosis. Results from this study suggest that interference of single metabolic enzyme activity altered multiple cellular signaling pathways.