Discovery of Specific Metastasis-Related N-Glycan Alterations in Epithelial Ovarian Cancer Based on Quantitative Glycomics

Zhang, Xingwang; Wang, Yisheng; Qian, Yifan; Wu, Xin; Zhang, Zejian; Liu, Xijun; Zhao, Ran; Zhou, Lei; Ruan, Yuanyuan; Xu, Jiejie; Liu, Haiou; Ren, Shifang; Xu, Congjian; Gu, Jianxin

doi:10.1371/journal.pone.0087978

Cited by 45 publications

(50 citation statements)

References 79 publications

(107 reference statements)

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“…Importantly, no other glycosyltransferase was shown to contribute to the formation of bisecting GlcNAc containing N -glycans as shown through genetic knockout studies in mice [7, 8]. In regards to ovarian cancer, the presence of this glycan motif was described in membrane and secreted glycoproteins from serous ovarian cancer specimens [9, 10] and MGAT3 expression was shown to be elevated in ovarian cancer cell lines and tissue when compared to normal counterparts [11, 12]. …”

Section: Introductionmentioning

confidence: 99%

Epigenetic activation of MGAT3 and corresponding bisecting GlcNAc shortens the survival of cancer patients

et al. 2016

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Bisecting GlcNAc on N-glycoproteins is described in E-cadherin-, EGF-, Wnt- and integrin- cancer-associated signaling pathways. However, the mechanisms regulating bisecting GlcNAc expression are not clear. Bisecting GlcNAc is attached to N-glycans through beta 1-4 N-acetylglucosaminyl transferase III (MGAT3), which is encoded by two exons flanked by high-density CpG islands. Despite a recently described correlation of MGAT3 and bisecting GlcNAc in ovarian cancer cells, it remains unknown whether DNA methylation is causative for the presence of bisecting GlcNAc. Here, we narrow down the regulatory genomic region and show that reconstitution of MGAT3 expression with 5-Aza coincides with reduced DNA methylation at the MGAT3 transcription start site. The presence of bisecting GlcNAc on released N-glycans was detected by mass spectrometry (LC-ESI-qTOF-MS/MS) in serous ovarian cancer cells upon DNA methyltransferase inhibition. The regulatory impact of DNA methylation on MGAT3 was further evaluated in 18 TCGA cancer types (n = 6118 samples) and the results indicate an improved overall survival in patients with reduced MGAT3 expression, thereby identifying long-term survivors of high-grade serous ovarian cancers (HGSOC). Epigenetic activation of MGAT3 was also confirmed in basal-like breast cancers sharing similar molecular and genetic features with HGSOC. These results provide novel insights into the epigenetic regulation of MGAT3/bisecting GlcNAc and demonstrate the importance of N-glycosylation in cancer progression.

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Section: Introductionmentioning

confidence: 99%

Epigenetic activation of MGAT3 and corresponding bisecting GlcNAc shortens the survival of cancer patients

et al. 2016

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“…It was also shown that the N-glycan-dependent mesothelin- MUC16 binding facilitates EOC peritoneal dissemination [39]. Two recent studies have described metastasis-related N-glycan alterations in EOC secretomes in vitro [40, 41], as definite N-glycan substructures and their complexes were found to be associated with specific epigenetic programming of synthetic enzymes in EOC [41]. Moreover, specific sialylated N-glycoproteins were exclusively identified in biological fluids from EOC patients [42].…”

Section: Discussionmentioning

confidence: 99%

A metabolic labeling approach for glycoproteomic analysis reveals altered glycoprotein expression upon GALNT3 knockdown in ovarian cancer cells

Sheta

Woo

Roux‐Dalvai³

et al. 2016

Journal of Proteomics

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Epithelial ovarian cancer (EOC) is a disease responsible for more deaths among women in the Western world than all other gynecologic malignancies. There is urgent need for new therapeutic targets and a better understanding of EOC initiation and progression. We have previously identified the polypeptide N-acetylgalactosaminyltransferase 3 (GALNT3) gene, a member of the GalNAc-transferases (GalNAc-Ts) gene family, as hypomethylated and overexpressed in high-grade serous EOC tumors, compared to low malignant potential EOC tumors and normal ovarian tissues. This data also suggested for a role of GALNT3 in aberrant EOC glycosylation, possibly implicated in disease progression. To evaluate differential glycosylation in EOC caused by modulations in GALNT3 expression, we used a metabolic labeling strategy for enrichment and mass spectrometry-based characterization of glycoproteins following GALNT3 gene knockdown (KD) in A2780s EOC cells. A total of 589 differentially expressed glycoproteins were identified upon GALNT3 KD. Most identified proteins were involved in mechanisms of cellular metabolic functions, post-translational modifications, and some have been reported to be implicated in EOC etiology. The GALNT3-dependent glycoproteins identified by this metabolic labeling approach support the oncogenic role of GALNT3 in EOC dissemination and may be pursued as novel EOC biomarkers and/or therapeutic targets. Biological significance: Knowledge of the O-glycoproteome has been relatively elusive, and the functions of the individual polypeptide GalNAc-Ts have been poorly characterized. Alterations in GalNAc-Ts expression were shown to provide huge variability in the O-glycoproteome in various pathologies, including cancer. The application of a chemical biology approach for the metabolic labeling and subsequent characterization of O-glycoproteins in EOC using the Ac4GalNAz metabolite has provided a strategy allowing for proteomic discovery of GalNAc-Ts specific functions. Our study supports an essential role of one of the GalNAc-Ts — GALNT3, in EOC dissemination, including its implication in modulating PTMs and EOC metabolism. Our approach validates the use of the applied metabolic strategy to identify important functions of GalNAc-Ts in normal and pathological conditions.

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“…The N-glycans in solution were purified and desalted using a Porous Graphic Carbon Solid-Phase Extraction (PGC-SPE) as previously described [21]. The PGC-SPE microcolumn was a GELoader tip filled with porous graphic carbon powder.…”

Section: Methodsmentioning

confidence: 99%

Integrated glycomic analysis of ovarian cancer side population cells

et al. 2016

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BackgroundOvarian cancer is the most lethal gynecological malignancy due to its frequent recurrence and drug resistance even after successful initial treatment. Accumulating scientific evidence indicates that subpopulations of cancer cells with stem cell-like properties, such as so-called side population (SP) cells, are primarily responsible for these recurrences. A better understanding of SP cells may provide new clues for detecting and targeting these cancer-initiating cells and ultimately help to eradicate cancer. Changes in glycosylation patterns are remarkable features of SP cells. Here, we isolated SP cells from ovarian cancer cell lines and analyzed their glycosylation patterns using multiple glycomic strategies.MethodsSix high-grade serous ovarian cancer cell lines were used for SP cell isolation. Among them, HO8910 pm, which contained the highest proportion of SP cells, was used for glycomic analysis of SP cells. Cell lysate of SP cells and main population cells was applied to lectin microarray and mass spectrometry for glycan profiling. Differently expressed glycan structures were further verified by lectin blot, flow cytometry, and real-time PCR analysis of their relevant enzymes.ResultsExpression of core fucosylated N-glycan and tumor-associated Tn, T and sT antigens were increased in SP cells. By contrast, SP cells exhibited decreased hybrid glycan, α2,3-linked sialic glycan and multivalent sialyl-glycan.ConclusionsGlycan structures, such as Tn, T, sT antigens, and core fucosylation may serve as biomarkers of ovarian cancer stem cells.Electronic supplementary materialThe online version of this article (doi:10.1186/s12014-016-9131-z) contains supplementary material, which is available to authorized users.

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Discovery of Specific Metastasis-Related N-Glycan Alterations in Epithelial Ovarian Cancer Based on Quantitative Glycomics

Cited by 45 publications

References 79 publications

Epigenetic activation of MGAT3 and corresponding bisecting GlcNAc shortens the survival of cancer patients

Epigenetic activation of MGAT3 and corresponding bisecting GlcNAc shortens the survival of cancer patients

A metabolic labeling approach for glycoproteomic analysis reveals altered glycoprotein expression upon GALNT3 knockdown in ovarian cancer cells

Integrated glycomic analysis of ovarian cancer side population cells

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