Dynamic analysis of N-glycomic and transcriptomic changes in the development of ovarian cancer cell line A2780 to its three cisplatin-resistant variants
Dynamic analysis of N-glycomic and transcriptomic changes in the development of ovarian cancer cell line A2780 to its three cisplatin-resistant variants
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卵巢癌细胞系 A2780 向其三种顺铂耐药变体发展过程中 N 糖组和转录组变化的动态分析。
DOI：
10.21037/atm.2020.03.12
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发表时间：
2020-03-01
影响因子：
--
通讯作者：
Xu, Congjian
中科院分区：
文献类型：
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作者：
Lin, Guiling;Zhao, Ran;Xu, Congjian
背景：铂类耐药的发展是连续化疗过程中发生的动态过程，导致卵巢癌的高死亡率。据报道，铂耐药性存在异常糖基化。已经进行了许多关于铂耐药性的研究，但很少有研究基于 N-糖组学研究与铂耐药性相关的聚糖。此外，卵巢癌铂类耐药发展过程中的血糖变化很少有报道。因此，本研究的目的是确定连续暴露于顺铂期间卵巢癌细胞中与铂耐药相关的 N-聚糖。这些聚糖可能参与铂耐药机制，并作为监测其发展的生物标志物。方法：本研究通过将A2780细胞持续暴露于顺铂来模拟卵巢癌铂耐药的发展。与 A2780 细胞相比，顺铂耐药变体通过较高的半数抑制浓度 (IC50) 值和增加的 P-糖蛋白 (ABCB1、P-gp) 表达得到证实。使用 MALDI 飞行时间 (TOF)-MS 结合乙酯化衍生化分析顺铂耐药变体在铂耐药性发展过程中的动态 N-糖组变化，用于区分 α 2,3- 和 α 2,6- 连接 N-乙酰神经氨酸。通过转录组测序和实时 PCR (RT-PCR) 在糖转移酶水平上进一步验证了 N-聚糖的改变。结果：与 A2780 细胞相比，MS 分析表明，在三种顺铂耐药变体中，α2,3-连接唾液酸结构和 N-聚糖 gal 比率显着较高，而岩藻糖基化聚糖较低。转录组测序和RT-PCR显示，三种顺铂耐药变异体中ST3GAL6和MGAT4A的基因表达增加，而FUT11、FUT1、GMDS和B4GALT5的基因表达减少。结论：N-聚糖和糖原表达分析表明，α2,3-连接的唾液酸结构可以作为监测铂类耐药发展的生物标志物，并指导卵巢癌患者的个体化治疗。
Background: Platinum resistance development is a dynamic process that occurs during continuous chemotherapy and contributes to high mortality in ovarian cancer. Abnormal glycosylation has been reported in platinum resistance. Many studies on platinum resistance have been performed, but few of them have investigated platinum resistance-associated glycans based on N-glycomics. Moreover, glycomic alterations during platinum resistance development in ovarian cancer are rarely reported. Therefore, the objective of this study was to determine platinum resistance-related N-glycans in ovarian cancer cells during continuous exposure to cisplatin. These glycans might be involved in the mechanism of platinum resistance and serve as biomarkers to monitor its development.Methods: This study mimicked the development of platinum resistance in ovarian cancer by continuously exposing A2780 cells to cisplatin. Cisplatin-resistant variants were confirmed by higher half maximal inhibitory concentration (IC50) values and increased P-glycoprotein (ABCB1, P-gp) expression compared to A2780 cells. Analysis of dynamic N-glycomic changes during the development of platinum resistance in cisplatin-resistant variants was performed with MALDI-time-of-flight (TOF)-MS combined with ethyl esterification derivatization, which were used to discriminate between alpha 2,3- and alpha 2,6-linkage N-acetylneuraminic acid. N-glycan alterations were further validated on a glycotransferase level via transcriptome sequencing and real-time PCR (RT-PCR).Results: Compared to the A2780 cells, MS analysis indicated that alpha 2,3-linked sialic structures and N-glycan gal-ratios were significantly higher, while fucosylated glycans were lower in three cisplatin-resistant variants. Transcriptome sequencing and RT-PCR showed that gene expression of ST3GAL6 and MGAT4A increased, while gene expression of FUT11, FUT1, GMDS, and B4GALT5 decreased in three cisplatin-resistant variants.Conclusions: Analysis of N-glycans and glycogene expression showed that alpha 2,3-linked sialic structures might serve as biomarkers to monitor the development of platinum resistance and to guide individualized treatment of ovarian cancer patients.