Glycome Profiling of Cancer Cell Lines Cultivated in Physiological and Commercial Media

Wang, Junyao; Peng, Wenjing; Yu, Aiying; Fokar, Mohamed; Mechref, Yehia

doi:10.3390/biom12060743

Cited by 4 publications

(3 citation statements)

References 56 publications

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“…46 Tandem MS (MS/MS) is frequently employed for the complete and accurate characterization of glycans and glycoproteins. Thus, LC-MS/MS-based glycoproteomic and glycomics analyses have gained significant attention in clinical research and have broadened our understanding of disease progression in various cancer forms, including lung, 47 brain, 48 and breast cancer. 17 In this study, O-glycans derived from five breast cancer and one brain cancer cell lines were chemoenzymatically released and analyzed by LC-MS/MS.…”

Section: Introductionmentioning

confidence: 99%

O-Glycome Profiling of Breast Cancer Cell Lines to Understand Breast Cancer Brain Metastasis

Onigbinde,

Peng,

Reddy

et al. 2024

J. Proteome Res.

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Breast cancer is the second leading cause of cancer-related death among women and a major source of brain metastases. Despite the increasing incidence of brain metastasis from breast cancer, the underlying mechanisms remain poorly understood. Altered glycosylation is known to play a role in various diseases including cancer metastasis. However, profiling studies of O-glycans and their isomers in breast cancer brain metastasis (BCBM) are scarce. This study analyzed the expression of O-glycans and their isomers in human breast cancer cell lines (MDA-MB-231, MDA-MB-361, HTB131, and HTB22), a brain cancer cell line (CRL-1620), and a brain metastatic breast cancer cell line (MDA-MB-231BR) using nanoLC-MS/MS, identifying 27 O-glycan compositions. We observed significant upregulation in the expression of HexNAc 1 Hex 1 NeuAc 2 and HexNAc 2 Hex 3 , whereas the expression of HexNAc 1 Hex 1 NeuAc 1 was downregulated in MDA-MB-231BR compared to other cell lines. In our isomeric analysis, we observed notable alterations in the isomeric forms of the O-glycan structure HexNAc 1 Hex 1 NeuAc 1 in a comparison of different cell lines. Our analysis of Oglycans and their isomers in cancer cells demonstrated that changes in their distribution can be related to the metastatic process. We believe that our investigation will contribute to an enhanced comprehension of the significance of O-glycans and their isomers in BCBM.

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Section: Introductionmentioning

confidence: 99%

O-Glycome Profiling of Breast Cancer Cell Lines to Understand Breast Cancer Brain Metastasis

Onigbinde,

Peng,

Reddy

et al. 2024

J. Proteome Res.

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“…The analytical strategy for glycome profiling usually includes the extraction of glycans followed by instrumental analyses [23,24]. Enzymatic cleavage has been commonly used by researchers to remove glycans from glycoproteins.…”

Section: Introductionmentioning

confidence: 99%

An Efficient and Economical N-Glycome Sample Preparation Using Acetone Precipitation

et al. 2022

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Due to the critical role of the glycome in organisms and its close connections with various diseases, much time and effort have been dedicated to glycomics-related studies in the past decade. To achieve accurate and reliable identification and quantification of glycans extracted from biological samples, several analysis methods have been well-developed. One commonly used methodology for the sample preparation of N-glycomics usually involves enzymatic cleavage by PNGase F, followed by sample purification using C18 cartridges to remove proteins. PNGase F and C18 cartridges are very efficient both for cleaving N-glycans and for protein removal. However, this method is most suitable for a limited quantity of samples. In this study, we developed a sample preparation method focusing on N-glycome extraction and purification from large-scale biological samples using acetone precipitation. The N-glycan yield was first tested on standard glycoprotein samples, bovine fetuin and complex biological samples, and human serum. Compared to C18 cartridges, most of the sialylated N-glycans from human serum were detected with higher abundance after acetone precipitation. However, C18 showed a slightly higher efficiency for protein removal. Using the unfiltered human serum as the baseline, around 97.7% of the proteins were removed by acetone precipitation, while more than 99.9% of the proteins were removed by C18 cartridges. Lastly, the acetone precipitation was applied to N-glycome extraction from egg yolks to demonstrate large-scale glycomics sample preparation.

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“…In addition, the transcriptional and metabolic phenotypes of cancer cells cultivated by Plasmax were found independent of the proliferation rate, which suggests that the high concentrations of some nutrients in commercial media is not required for cell growth, and it misdirects cell line metabolism. Our previous study also suggests that Plasmax can lead to alterations in glycosylation [ 17 ]. However, a comprehensive investigation of gene and protein expression changes in Plasmax medium is necessary to guide the medium selection when performing in vitro cell line experiments.…”

Section: Introductionmentioning

confidence: 99%

Comparative Transcriptomics and Proteomics of Cancer Cell Lines Cultivated by Physiological and Commercial Media

Wang

Peng

et al. 2022

Biomolecules

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Aiming to reduce the gap between in vitro and in vivo environment, a complex culture medium, Plasmax, was introduced recently, which includes nutrients and metabolites with concentrations normally found in human plasma. Herein, to study the influence of this medium on cellular behaviors, we utilized Plasmax to cultivate two cancer cell lines, including one breast cancer cell line, MDA-MB-231BR, and one brain cancer cell line, CRL-1620. Cancer cells were harvested and prepared for transcriptomics and proteomics analyses to assess the discrepancies caused by the different nutritional environments of Plasmax and two commercial media: DMEM, and EMEM. Total RNAs of cells were extracted using mammalian total RNA extract kits and analyzed by next-generation RNA sequencing; proteomics analyses were performed using LC-MS/MS. Gene oncology and pathway analysis were employed to study the affected functions. The cellular invasion and cell death were inhibited in MDA-MB-231BR cell line when cultured in Plasmax compared to DMEM and EMEM, whereas the invasion, migration and protein synthesis of CRL-1620 cell line were activated in Plasmax in relative to both commercial media. The expression changes of some proteins were more significant compared to their corresponding transcripts, indicating that Plasmax has more influence upon regulatory processes of proteins after translation. This work provides complementary information to the original study of Plasmax, aiming to facilitate the selection of appropriate media for in vitro cancer cell studies.

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Glycome Profiling of Cancer Cell Lines Cultivated in Physiological and Commercial Media

Cited by 4 publications

References 56 publications

O-Glycome Profiling of Breast Cancer Cell Lines to Understand Breast Cancer Brain Metastasis

O-Glycome Profiling of Breast Cancer Cell Lines to Understand Breast Cancer Brain Metastasis

An Efficient and Economical N-Glycome Sample Preparation Using Acetone Precipitation

Comparative Transcriptomics and Proteomics of Cancer Cell Lines Cultivated by Physiological and Commercial Media

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