The lectin Helix pomatia agglutinin recognizes O-GlcNAc containing glycoproteins in human breast cancer
The lectin Helix pomatia agglutinin recognizes O-GlcNAc containing glycoproteins in human breast cancer
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DOI：
10.1093/glycob/cws051
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发表时间：
2012-06-01
期刊：
影响因子：
4.3
通讯作者：
Dwek, Miriam V.
中科院分区：
文献类型：
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作者：
Rambaruth, Neela D. S.;Greenwell, Pamela;Dwek, Miriam V.
由于凝集素已被证明能识别与转移性疾病发展相关的糖基化变化，人们对了解乳腺癌中凝集素螺旋凝集素(HPA)的表位非常感兴趣。HPA以前被证明识别癌症中异常的O-连接的α-N-乙酰半乳糖胺(GalNAcα)/粘蛋白糖基化，包括暴露的Tn表位。然而，最近的糖阵列分析报告，凝集素也识别不同的表位，例如功能糖组(CFG)数据联盟：GalNAcα1，3Gal；β-GalNAc；GlcNAcβ1，4Gal。来自CFG阵列的有趣观察导致了这项研究，其中HPA结合表位在乳腺癌转移的体外模型中被定位和表征。HMT3522(良性疾病)、BT474(原发癌)和T47D/MCF7(转移性癌)细胞进行基于共聚焦显微镜的共定位研究，并采用基于二维电泳(2DE)、Western blotting和质谱仪的糖蛋白组学分析。HPA结合与整合素α6、转录因子异质性核糖核蛋白(HnRNP)H1、hnRNP D样蛋白、hnRNP A2/B1以及热休克蛋白27(Hsp27)、胶质纤维酸性蛋白和烯醇化酶1(ENO1)的水平相关。这些糖蛋白在非转移性乳腺癌细胞系中检测不到。HPA对hnRNPs、Hsp27和ENO1的识别与这些蛋白的O-GlcN酰化有关。在具有转移表型的乳腺癌细胞中，整合素α6是最丰富的HPA糖蛋白；这与以前在结直肠癌中的发现一致。这是第一个HPA与O-GlcNacylated转录因子结合的报道。这类蛋白质代表了HPA区分具有侵袭性转移表型的癌细胞的新方法。
There has been considerable interest in understanding the epitopes that bind the lectin Helix pomatia agglutinin (HPA) in breast cancer as the lectin has been shown to identify glycosylation changes associated with the development of metastatic disease. HPA has previously been shown to recognize aberrant O-linked alpha-N-acetylgalactosamine (GalNAc alpha)/mucin glycosylation in cancer, including exposed Tn epitopes. However, recent glycan-array analysis reported that diverse epitopes are also recognized by the lectin, e.g. consortium for functional glycomics (CFG) data: GalNAc alpha 1,3Gal; beta-GalNAc; GlcNAc beta 1,4Gal. The intriguing observations from the CFG array led to this study, in which HPA-binding epitopes were localized and characterized in an in vitro model of breast cancer metastasis. HMT3522 (benign disease), BT474 (primary cancer) and T47D/MCF7 (metastatic cancer) cells were assessed in confocal microscopy-based co-localization studies and a glycoproteomic analysis based on 2-dimensional electrophoresis (2DE), western blotting and mass spectrometry was adopted. HPA binding correlated with levels of integrin alpha 6, transcription factors heterogeneous nuclear ribonuclear protein (HnRNP) H1, HnRNP D-like, HnRNP A2/B1 as well as heat shock protein 27 (Hsp27), glial fibrillary acidic protein and enolase 1 (ENO1). These glycoproteins were non-detectable in the non-metastatic breast cancer cell lines. The recognition of HnRNPs, Hsp27 and ENO1 by HPA correlated with O-GlcNAcylation of these proteins. Integrin alpha 6 was the most abundant HPA glycoprotein in the breast cancer cells with a metastatic phenotype; this concurred with previous findings in colorectal cancer. This is the first report in which HPA has been shown to bind O-GlcNAcylated transcription factors. This class of proteins represents a new means by which HPA differentiates cancer cells with an aggressive metastatic phenotype.