基于转录组与代谢组学联合分析谷氨酰胺合成酶在慢性髓系白血病耐药中的作用机制

doi:10.11958/20252219

天津医药 ›› 2026, Vol. 54 ›› Issue (1): 1-7.doi: 10.11958/20252219

• 细胞与分子生物学 • 下一篇

基于转录组与代谢组学联合分析谷氨酰胺合成酶在慢性髓系白血病耐药中的作用机制

庄瑞玲(), 杨杰, 陈艳欣, 张雲^△()

中国人民解放军联勤保障部队第九〇〇医院血液科（邮编350000）

收稿日期:2025-06-06 修回日期:2025-09-21 出版日期:2026-01-15 发布日期:2026-01-19
通讯作者: ^△ E-mail：306005288@qq.com
作者简介:庄瑞玲（1994），女，主治医师，主要从事中西医结合治疗血液病方面研究。E-mail：18016721917@163.com
基金资助:
福建省自然科学基金资助项目(2024J01583)

Based on the combined transcriptomic and metabolomic analysis of the regulatory role of GLUL in drug resistance in chronic myeloid leukemia

ZHUANG Ruiling(), YANG Jie, CHEN Yanxin, ZHANG Yun^△()

Department of Hematology, 900th Hospital of PLA Joint Logistic Support Force, Fuzhou 350000, China

Received:2025-06-06 Revised:2025-09-21 Published:2026-01-15 Online:2026-01-19
Contact: ^△ E-mail：306005288@qq.com

庄瑞玲, 杨杰, 陈艳欣, 张雲. 基于转录组与代谢组学联合分析谷氨酰胺合成酶在慢性髓系白血病耐药中的作用机制[J]. 天津医药, 2026, 54(1): 1-7.

ZHUANG Ruiling, YANG Jie, CHEN Yanxin, ZHANG Yun. Based on the combined transcriptomic and metabolomic analysis of the regulatory role of GLUL in drug resistance in chronic myeloid leukemia[J]. Tianjin Medical Journal, 2026, 54(1): 1-7.

摘要/Abstract

摘要：

目的探讨白血病K562细胞与耐药K562-G01细胞在基因表达和代谢组学上的差异，并分析谷氨酰胺合成酶（GLUL）在调控K562-G01耐药性中的潜在作用。方法体外培养K562与耐药K562-G01细胞，通过转录组学和代谢组学筛选耐药细胞中的差异表达基因、差异代谢物及相关通路，通过关联分析确定转录组与代谢组共同富集的信号通路。通过qPCR和Western blot检测差异基因的表达，流式细胞术评估GLUL基因干扰对细胞凋亡的影响。结果转录组分析结果显示，K562-G01细胞中有2 423个基因表达上调，3 321个基因表达下调。代谢组学分析显示，K562-G01中570个代谢物表达上调，779个表达下调。转录组与代谢组关联分析表明，丙氨酸、天冬氨酸和谷氨酸代谢通路参与了K562细胞耐药性的产生。qPCR和Western blot显示，在K562-G01细胞中GLUL的mRNA和蛋白表达上调（P<0.05），进一步发现，干扰GLUL的表达促进了K562和K562-G01细胞的凋亡，并增强了伊马替尼的药物敏感性（P<0.05）。结论丙氨酸、天冬氨酸和谷氨酸代谢通路的激活可能参与了K562-G01耐药，干扰GLUL可显著促进耐药细胞凋亡并增强化疗药物的敏感性。

关键词: 白血病, 髓系, 慢性, BCR-ABL阳性；抗药性, 肿瘤；计算生物学；谷氨酰胺合成酶

Abstract:

Objective To investigate the differences of gene expression and metabolomics between K562 cells and their resistant counterpart, K562-G01 cells, and analyze the potential role of glutamate-ammonia ligase (GLUL) in regulating the drug resistance of K562-G01 cells. Methods K562 and resistant K562-G01 cells were cultured in vitro. Differentially expressed genes, metabolites and related pathways in the resistant cells were identified through transcriptomics and metabolomics. The association between transcriptomic and metabolomic pathways was analyzed to identify common signaling pathways. Differential gene expression was detected by qPCR and Western blot assay. Flow cytometry was employed to evaluate the effect of GLUL gene interference on cell apoptosis. Results Transcriptomic analysis showed that 2 423 genes were upregulated, and 3 321 genes were downregulated in K562-G01 cells. Metabolomic analysis revealed that 570 metabolites were upregulated and 779 were downregulated in K562-G01 cells. Transcriptomic and metabolomic association analysis indicated that alanine, aspartate and glutamate metabolism pathway were involved in the development of drug resistance in K562 cells. qPCR and Western blot results demonstrated that GLUL expression was upregulated in K562-G01 cells (P<0.05). Further investigation showed that interfering with GLUL expression promoted apoptosis in both K562 and K562-G01 cells and enhanced the drug sensitivity to imatinib (P<0.05). Conclusion Activation of alanine, aspartate and glutamate metabolism pathways may contribute to the drug resistance of K562-G01 cells. Interfering with GLUL expression significantly promotes apoptosis in drug resistant cells and enhances the sensitivity to chemotherapy drugs.

Key words: leukemia, myelogenous, chronic, BCR-ABL positive, drug resistance, neoplasm, computational biology, glutamate-ammonia ligase

中图分类号:

R733.722

图/表 9

表1 引物序列和干扰序列

Tab.1 Primer sequences and interference sequences

基因名称	引物/干扰序列（5′→3′）	产物大小/bp
Homo β-actin	上游：CCCTGGAGAAGAGCTACGAG	180
Homo β-actin	下游：CGTACAGGTCTTTGCGGATG	180
Homo GLUL	上游：ATTACTGTGGTGTGGGAGCA	200
Homo GLUL	下游：ACGATGCAAGATGAAACGGG	200
NC	UUCUCCGAACGUGUCACGUTT-
NC	ACGUGACACGUUCGGAGAATT
si-GLUL-1	ATTCCATGAAACCTCCAACATT-
si-GLUL-1	TGTTGGAGGTTTCATGGAATTT
si-GLUL-2	GCTGCCATGTTTCGGGACCCCTTTT-
si-GLUL-2	TGTTGGAGGTTTCATGGAATTT
si-GLUL-3	CTGCGACCCCTTTTCGGTGACTT-
si-GLUL-3	GTCACCGAAAAGGGGTCGCAGTT

图1 K562和K562-G01细胞的DEGs筛选 A：火山图展示2组的DEGs；B：热图展示2组差异显著的基因。

Fig.1 Differential gene analysis between K562 and K562-G01 cells

图2 DEGs的通路富集分析 A：DEGs的GO功能注释；B：DEGs的KEGG信号通路注释。

Fig.2 Pathway enrichment analysis of differentially expressed genes

图3 差异代谢物的筛选及通路富集分析 A：火山图展示差异显著的代谢物；B：KEGG通路富集分析展示差异代谢物富集的信号通路。

Fig.3 Screening of differential metabolites and pathway enrichment analysis

图4 转录组和代谢组学结果关联分析 A：转录组和代谢组中差异基因和代谢物富集相同的信号通路；B：CCA图，横、纵坐标分别表示不同组学物质与典型变量U1和典型变量U2之间的简单相关系数，紫色、红色点分别表示代谢物和基因。图中以十字区分出4个区域，在同一个区域内，距原点越远，表示与典型变量关联性越强；相互距离越近，表示两者可能参与同一个生物学过程。

Fig.4 Correlation analysis of transcriptomic and metabolomic results

表2 K562和K562-G01细胞中的差异基因和代谢物比较

Tab.2 Differentially expressed genes and metabolites between K562 and K562 G01 cells ($\bar{x}±s$)

细胞	n	基因FPKM
细胞	n	ALDH4A1	GLUL	ADSS2	GFPT2
K562	3	15.71±4.08	74.28±4.56	7.96±2.16	0.04±0.02
K562-G01	3	24.68±0.24	110.71±2.72	19.45±3.32	0.19±0.09
t		4.509^＊	3.699^＊	2.884^＊	2.823^＊
细胞	n	代谢物丰度
细胞	n	琥珀酸半醛	N-乙酰天冬氨酰谷氨酸	γ-氨基丁酸	5-磷酸核糖胺
K562	6	6 455.00±1 081.87	68 900.00±141.42	514.00±1.41	6 495.00±742.46
K562-G01	6	32 000.00±21 071.78	22 400.00±27 152.90	8 950.00±3 747.67	46 500.00±24 041.63
t		3.976^＊＊	3.987^＊＊	3.204^＊＊	4.188^＊＊

图5 Western blot检测K562和K562-G01细胞中GLUL的表达水平以及干扰效率 A：Western blot检测GLUL在K562和K562-G01细胞中的表达水平；B：Western blot检测si-GLUL在K562中的干扰效率；C：Western blot检测si-GLUL在K562-G01细胞中的干扰效率。

Fig.5 Western blot detection of GLUL expression level in K562 and K562 drug-resistant cells and interference efficiency

表3 GLUL的干扰效率检测结果

Tab.3 Detection results of interference efficiency of GLUL （n=3，$\bar{x}±s$）

组别	GLUL基因		GLUL蛋白
组别	K562	K562-G01	K562	K562-G01
对照组	1.00±0.00	1.00±0.00	0.66±0.07	0.88±0.24
si-GLUL-1组	0.81±0.19	0.83±0.21	0.71±0.03	0.86±0.01
si-GLUL-2组	0.44±0.12^ab	0.39±0.13^ab	0.34±0.07^ab	0.49±0.05^ab
si-GLUL-3组	0.64±0.03^ac	0.62±0.08^ac	0.52±0.20^ac	0.73±0.03^ac
F	24.112^＊＊	18.443^＊＊	19.695^＊＊	87.524^＊＊

表4 各组细胞凋亡率比较

Tab.4 Comparison of apoptosis rates between the five groups of cells （n=3，%，$\bar{x}±s$）

组别	K562细胞	K562-G01细胞
NC组	10.17±1.16	10.90±0.61
si-GLUL组	26.86±1.40^a	17.13±1.22^a
伊马替尼组	20.30±0.84^a	15.15±1.37^a
si-GLUL+伊马替尼组	43.71±2.23^bc	32.16±1.94^bc
F	265.300^＊＊	137.000^＊＊

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基于转录组与代谢组学联合分析谷氨酰胺合成酶在慢性髓系白血病耐药中的作用机制

Based on the combined transcriptomic and metabolomic analysis of the regulatory role of GLUL in drug resistance in chronic myeloid leukemia

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