Proteomic analysis and abrogated expression of O-GlcNAcylated proteins associated with primary breast cancer
Proteomic analysis and abrogated expression of O-GlcNAcylated proteins associated with primary breast cancer
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DOI：
10.1002/pmic.201200126
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发表时间：
2013-07-01
期刊：
影响因子：
3.4
通讯作者：
Svasti, Jisnuson
中科院分区：
文献类型：
--
作者：
Champattanachai, Voraratt;Netsirisawan, Pukkavadee;Svasti, Jisnuson
O-GlcNAc化是细胞核和细胞质蛋白的动态PTM，由O-GlcNAc转移酶（OGT）和O-GlcNAc酶调节，O-GlcNAc转移酶和O-GlcNAc酶分别催化O-GlcNAc的添加和去除。这种修饰与葡萄糖代谢有关，葡萄糖代谢在包括癌症在内的许多疾病中起着重要作用。虽然新出现的证据表明，一些肿瘤相关蛋白质是O-GlcNAc修饰的，但癌症中的总O-GlcNAc化在很大程度上仍未被探索。在这里，我们证明了O-GlcNAc化在原发性乳腺恶性肿瘤中增加，而不是在良性肿瘤中，并且这种增强与OGT水平的表达增加相关。使用2D O-GlcNAc免疫印迹和LC-MS/MS分析，我们成功地鉴定了29种蛋白质，其中7种是独特的O-GlcNAc酰化或与癌症中的O-GlcNAc酰化相关。在这些鉴定的蛋白质中，一些与瓦尔堡效应相关，包括代谢酶、参与胁迫反应和生物合成的蛋白质。此外，与RNA代谢、基因表达和细胞骨架相关的蛋白质被高度O-GlcNAc化或与O-GlcNAc化相关。此外，OGT敲低表明O-GlcNAc化的降低与体外抑制非锚定依赖性生长有关。这些数据表明异常蛋白O-GlcNAc化与乳腺癌相关。这些O-GlcNAc修饰蛋白的异常修饰可能是肿瘤的重要恶性特征之一。
O-GlcNAcylation is a dynamic PTM of nuclear and cytoplasmic proteins, regulated by O-GlcNAc transferase (OGT) and O-GlcNAcase, which catalyze the addition and removal of O-GlcNAc, respectively. This modification is associated with glucose metabolism, which plays important roles in many diseases including cancer. Although emerging evidence reveals that some tumor-associated proteins are O-GlcNAc modified, the total O-GlcNAcylation in cancer is still largely unexplored. Here, we demonstrate that O-GlcNAcylation was increased in primary breast malignant tumors, not in benign tumors and that this augmentation was associated with increased expression of OGT level. Using 2D O-GlcNAc immnoblotting and LC-MS/MS analysis, we successfully identified 29 proteins, with seven being uniquely O-GlcNAcylated or associated with O-GlcNAcylation in cancer. Of these identified proteins, some were related to the Warburg effect, including metabolic enzymes, proteins involved in stress responses and biosynthesis. In addition, proteins associated with RNA metabolism, gene expression, and cytoskeleton were highly O-GlcNAcylated or associated with O-GlcNAcylation. Moreover, OGT knockdown showed that decreasingO-GlcNAcylation was related to inhibition of the anchorage-independent growth in vitro. These data indicate that aberrant protein O-GlcNAcylation is associated with breast cancer. Abnormal modification of these O-GlcNAc-modified proteins might be one of the vital malignant characteristics of cancer.