Department of Chemistry and Biochemistry, Texas Tech University, Lubbock, TX 79409, USA.
Center of Biotechnology and Genomics, Texas Tech University, Lubbock, TX 79409, USA.
Biomolecules. 2022 May 24;12(6):743. doi: 10.3390/biom12060743.
A complex physiological culture medium (Plasmax) was introduced recently, composed of nutrients and metabolites at concentrations normally found in human plasma to mimic the in vivo environment for cell line cultivation. As glycosylation has been proved to be involved in cancer development, it is necessary to investigate the glycan expression changes in media with different nutrients. In this study, a breast cancer cell line, MDA-MB-231BR, and a brain cancer cell line, CRL-1620, were cultivated in Plasmax and commercial media to reveal cell line glycosylation discrepancies prompted by nutritional environments. Glycomics analyses of cell lines were performed using LC-MS/MS. The expressions of multiple fucosylated -glycans, such as HexNAcHexDeoxyHex and HexNAcHexDeoxyHex, derived from both cell lines exhibited a significant increase in Plasmax. Among the -glycans, significant differences were also observed. Both cell lines cultivated in EMEM had the lowest amounts of -glycans expressed. The original work described the development of Plasmax, which improves colony formation, and resulted in transcriptomic and metabolomic alterations of cancer cell lines, while our results indicate that Plasmax can significantly impact protein glycosylation. This study also provides information to guide the selection of media for in vitro cancer cell glycomics studies.
最近引入了一种复杂的生理培养基（Plasmax），它由人类血浆中正常存在的营养物质和代谢物组成，浓度与体内环境相似，用于模拟细胞系的培养环境。由于糖基化已被证明与癌症的发展有关，因此有必要研究不同营养物质的培养基中聚糖表达的变化。在这项研究中，我们培养了乳腺癌细胞系 MDA-MB-231BR 和脑癌细胞系 CRL-1620，在 Plasmax 和商业培养基中培养，以揭示营养环境引起的细胞系糖基化差异。使用 LC-MS/MS 对细胞系进行了糖组学分析。来自两种细胞系的多个岩藻糖基化的聚糖，如 HexNAcHexDeoxyHex 和 HexNAcHexDeoxyHex 的表达在 Plasmax 中显著增加。在这些聚糖中，也观察到了显著的差异。在 EMEM 中培养的两种细胞系表达的聚糖量最低。原始工作描述了 Plasmax 的发展，它提高了集落形成能力，并导致癌细胞系的转录组和代谢组发生改变，而我们的结果表明 Plasmax 可以显著影响蛋白质糖基化。本研究还为指导体外癌症细胞糖组学研究中培养基的选择提供了信息。
