过表达或抑制细胞周期蛋白依赖性激酶2对A375细胞代谢组的影响

doi:10.35541/cjd.20240442

中华皮肤科杂志 ›› 2025, e20240442.doi: 10.35541/cjd.20240442

过表达或抑制细胞周期蛋白依赖性激酶2对A375细胞代谢组的影响

刘厚广¹ 申宜² 魏琼玲¹ 李峥¹ 霍姗姗¹ 谢广成² 刘英文²

¹厦门市第三医院皮肤科，厦门 361100；²承德医学院基础医学院，承德 067000

收稿日期:2024-08-19 修回日期:2024-11-29 发布日期:2025-05-20
通讯作者: 刘英文 E-mail:LLY1989218@126.com
基金资助:
福建省卫生健康科技计划项目（2021CXB027）

Effect of overexpression or inhibition of cyclin-dependent kinase 2 on the metabolome of A375 cells

Liu Houguang¹, Shen Yi², Wei Qiongling¹, Li Zheng¹, Huo Shanshan¹, Xie Guangcheng², Liu Yingwen²

¹Department of Dermatology, the Third Hospital of Xiamen, Xiamen 361100, China; ²College of Basic Medicine, Chengde Medical University, Chengde 067000, China

Received:2024-08-19 Revised:2024-11-29 Published:2025-05-20
Contact: Liu Yingwen E-mail:LLY1989218@126.com
Supported by:
Fujian Provincial Health Technology Project（2021CXB027）

摘要/Abstract

摘要： 【摘要】目的研究过表达或抑制细胞周期蛋白依赖性激酶2（CDK2）对人黑色素瘤细胞代谢组的影响。方法将重组真核表达质粒pcDNA3.1（+）-CDK2+myc转染至人黑色素瘤A375细胞进行CDK2过表达；使用0（对照）、0.9和1.8 μmol/L的CDK2抑制剂Roscovitine处理A375细胞24 h；免疫印迹检测CDK2蛋白表达情况。采用非靶向代谢组学检测过表达或抑制CDK2后A375细胞的代谢组改变情况，确定差异显著代谢产物（DSM）的数量和类型，并进行DSM的相关性、京都基因与基因组百科全书（KEGG）通路和代表性代谢通路的聚类分析。结果转染重组真核表达质粒pcDNA3.1（+）-CDK2+myc能够提升A375细胞CDK2表达量；0.9和1.8 μmol/L Roscovitine能降低CDK2的表达量。过表达CDK2时，共检测出483个DSM；而抑制CDK2时，共检测出306个DSM；DSM以下调表达为主。有机酸及其衍生物、有机杂环化合物、苯环型化合物、核苷类核苷酸及其类似物等DSM之间以及与未定义DSM之间存在相关性。DSM主要涉及到氨基酸代谢、脂质代谢等代谢途径。氨基酸的生物合成、蛋白质的消化和吸收和氨酰-tRNA生物合成途径为过表达或抑制CDK2共同调控代谢途径，且涉及到的DSM有所不同。结论过表达或抑制CDK2能显著改变A375细胞的代谢组。

关键词: 黑色素瘤, 实验性, 细胞周期蛋白质依赖激酶类, 代谢组学, A375细胞

Abstract: 【Abstract】 Objective To investigate the effect of overexpression or inhibition of cyclin-dependent kinase 2 （CDK2） on the metabolome of human melanoma cells. Methods The recombinant eukaryotic expression plasmid pcDNA3.1（+）-CDK2+myc was transfected into A375 human melanoma cells for CDK2 overexpression; A375 cells were divided into 3 groups to be treated with the CDK2 inhibitor Roscovitine at final concentrations of 0 （control）, 0.9 and 1.8 μmol/L for 24 hours; Western blot analysis was performed to determine the CDK2 protein expression. An untargeted metabolomics approach was used to investigate metabolomic changes in A375 cells after overexpression or inhibition of CDK2. The number and type of differentially significant metabolites （DSMs）were determined. Correlation analysis, Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway analysis and clustering analysis of representative metabolic pathways were carried out. Results The CDK2 expression level was significantly up-regulated in A375 cells after transfection with the pcDNA3.1（+）-CDK2+myc plasmid, whereas the treatment with 0.9 and 1.8 μmol/L Roscovitine down-regulated its expression. A total of 483 DSMs were identified in CDK2-overexpressed A375 cells, and 306 DSMs were identified in CDK2-inhibited A375 cells; down-regulated DSMs constituted the majority among all DSMs. There were correlations among DSMs such as organic acids and their derivatives, organoheterocyclic compounds, benzenoids, nucleosides/nucleotides and their analogues, as well as correlations between defined and undefined DSMs. The DSMs were mainly involved in metabolic pathways, such as amino acid metabolism and lipid metabolism. The biosynthesis of amino acids, protein digestion and absorption, and aminoacyl-tRNA biosynthesis were common metabolic pathways regulated by overexpression or inhibition of CDK2. However, the types of DSMs involved in these pathways were different. Conclusion Overexpression or inhibition of CDK2 can significantly change the metabolome of A375 cells.

Key words: Melanoma, experimental, Cyclin-dependent kinases, Metabolomics, A375 cells

刘厚广申宜魏琼玲李峥霍姗姗谢广成刘英文. 过表达或抑制细胞周期蛋白依赖性激酶2对A375细胞代谢组的影响[J]. 中华皮肤科杂志, 2025,e20240442. doi:10.35541/cjd.20240442

Liu Houguang, Shen Yi, Wei Qiongling, Li Zheng, Huo Shanshan, Xie Guangcheng, Liu Yingwen. Effect of overexpression or inhibition of cyclin-dependent kinase 2 on the metabolome of A375 cells[J]. Chinese Journal of Dermatology,2025,e20240442. doi:10.35541/cjd.20240442

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过表达或抑制细胞周期蛋白依赖性激酶2对A375细胞代谢组的影响

Effect of overexpression or inhibition of cyclin-dependent kinase 2 on the metabolome of A375 cells

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