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代谢组学分析揭示了致瘤性和非致瘤性 Madin-Darby 犬肾(MDCK)细胞增殖之间的差异。

Metabolomics profiling reveals differences in proliferation between tumorigenic and non-tumorigenic Madin-Darby canine kidney (MDCK) cells.

机构信息

Gansu Technology Innovation Center of Animal Cell, Biomedical Research Center, Northwest Minzu University, Lanzhou, China.

Engineering Research Center of Key Technology and Industrialization of Cell-based Vaccine, Ministry of Education, Lanzhou, China.

出版信息

PeerJ. 2023 Sep 20;11:e16077. doi: 10.7717/peerj.16077. eCollection 2023.

Abstract

BACKGROUND

Madin-Darby canine kidney (MDCK) cells are a cellular matrix in the production of influenza vaccines. The proliferation rate of MDCK cells is one of the critical factors that determine the vaccine production cycle. It is yet to be determined if there is a correlation between cell proliferation and alterations in metabolic levels. This study aimed to explore the metabolic differences between MDCK cells with varying proliferative capabilities through the use of both untargeted and targeted metabolomics.

METHODS

To investigate the metabolic discrepancies between adherent cell groups (MDCK-M60 and MDCK-CL23) and suspension cell groups (MDCK-XF04 and MDCK-XF06), untargeted and targeted metabolomics were used. Utilizing RT-qPCR analysis, the mRNA expressions of key metabolites enzymes were identified.

RESULTS

An untargeted metabolomics study demonstrated the presence of 81 metabolites between MDCK-M60 and MDCK-CL23 cells, which were mainly affected by six pathways. An analysis of MDCK-XF04 and MDCK-XF06 cells revealed a total of 113 potential metabolites, the majority of which were impacted by ten pathways. Targeted metabolomics revealed a decrease in the levels of choline, tryptophan, and tyrosine in MDCK-CL23 cells, which was in accordance with the results of untargeted metabolomics. Additionally, MDCK-XF06 cells experienced a decrease in 5'-methylthioadenosine and tryptophan, while S-adenosylhomocysteine, kynurenine, 11Z-eicosenoic acid, 3-phosphoglycerate, glucose 6-phosphate, and phosphoenolpyruvic acid concentrations were increased. The mRNA levels of MAT1A, MAT2B, IDO1, and IDO2 in the two cell groups were all increased, suggesting that S-adenosylmethionine and tryptophan may have a significant role in cell metabolism.

CONCLUSIONS

This research examines the effect of metabolite fluctuations on cell proliferation, thus offering a potential way to improve the rate of MDCK cell growth.

摘要

背景

MDCK 细胞(Madin-Darby 犬肾细胞)是流感疫苗生产中的细胞基质。MDCK 细胞的增殖率是决定疫苗生产周期的关键因素之一。目前尚不清楚细胞增殖与代谢水平变化之间是否存在相关性。本研究旨在通过非靶向和靶向代谢组学来探讨具有不同增殖能力的 MDCK 细胞之间的代谢差异。

方法

为了研究贴壁细胞群(MDCK-M60 和 MDCK-CL23)和悬浮细胞群(MDCK-XF04 和 MDCK-XF06)之间的代谢差异,我们使用了非靶向和靶向代谢组学。利用 RT-qPCR 分析,鉴定了关键代谢物酶的 mRNA 表达。

结果

非靶向代谢组学研究表明,MDCK-M60 和 MDCK-CL23 细胞之间存在 81 种代谢物,主要受 6 条途径影响。对 MDCK-XF04 和 MDCK-XF06 细胞的分析显示,共有 113 种潜在代谢物,其中大多数受 10 条途径影响。靶向代谢组学显示 MDCK-CL23 细胞中的胆碱、色氨酸和酪氨酸水平降低,这与非靶向代谢组学的结果一致。此外,MDCK-XF06 细胞中的 5'-甲基硫代腺苷和色氨酸水平降低,而 S-腺苷同型半胱氨酸、犬尿氨酸、11Z-二十碳烯酸、3-磷酸甘油酸、葡萄糖 6-磷酸和磷酸烯醇丙酮酸浓度升高。两组细胞中 MAT1A、MAT2B、IDO1 和 IDO2 的 mRNA 水平均升高,提示 S-腺苷甲硫氨酸和色氨酸可能在细胞代谢中起重要作用。

结论

本研究考察了代谢物波动对细胞增殖的影响,从而为提高 MDCK 细胞生长速度提供了一种潜在途径。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/126e/10517658/78957df621ad/peerj-11-16077-g001.jpg

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