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《中华肺部疾病杂志》[ISSN:1006-6977/CN:61-1281/TN]

卷:

期数:

2025年02期

页码:

273-278

栏目:

论著

出版日期:

2025-04-25

文章信息/Info

Title:

Regulation of 1-methylhistidine and dimethylglycine in lung adenocarcinoma by circUBAP2L via hsa-let-7c/GAD1 axis

作者:

夏祯祎¹; 刘 懿¹; 丁俞成¹; 袁宏霞²; 李晓燕²; 陈 田²

404000 重庆,重庆大学附属三峡医院胸外科¹；404000 重庆,重庆大学附属三峡医院健康管理中心²

Author(s):

Xia Zhenyi¹;  Liu Yi¹;  Ding Yucheng¹;  Yuan Hongxia²;  Li Xiaoyan²;  Chen Tian².

¹Department of Thoracic Surgery, Chongqing University Three Gorges Hospital, Chongqing 404000, China; ²Health Management Center, Chongqing University Three Gorges Hospital, Chongqing 404000, China

关键词:

肺腺癌;  环状泛素相关蛋白样因子2;  hsa-let-7c;  谷氨酸脱羧酶1;  肿瘤代谢

Keywords:

Lung adenocarcinoma;  Circular ubiquitin associated protein 2-like;  Hsa-let-7c;  Glutamatedecarboxylase1;  tumor metabolism

分类号:

R734.2

DOI:

10.3877/cma.j.issn.1674-6902.2025.02.013

摘要:

目的 分析环状泛素相关蛋白样因子2(circular ubiquitin associated protein 2-like, circUBAP2L)、hsa-let-7c及靶基因谷氨酸脱羧酶1(glutamatedecarboxylase1, GAD1)在肺腺癌中的作用。方法 采用small-RNA测序发现hsa-let-7c差异表达,通过生物信息学、实时聚合酶链反应(polymerase chain reaction, PCR)分析hsa-let-7c上游基因circUBAP2L、下游靶基因GAD1的差异表达。采用双荧光素酶报告基因实验印证circUBAP2L与hsa-let-7c、hsa-let-7c与GAD1的调控关系。通过慢病毒构建A549肺癌细胞GAD1过表达模型,转录组学RNA测序(RNA sequencing, RNA-seq)和代谢组学液相色谱-质谱联用(liquid chromatography-mass spectroscopy, LC-MS)分析GAD1的差异基因及代谢物,通过多组学联合分析GAD1对肺腺癌细胞的氨基酸代谢影响。结果 实时PCR实验印证circUBAP2L在肺腺癌中表达上调(P<0.01); TCGA和GTEx数据库分析结果显示,肺腺癌组织中hsa-let-7c表达降低(P<0.0001),Small-RNA测序结果印证hsa-let-7c在肺腺癌中表达降低(P<0.01); 转录组测序显示与正常组织相比,GAD1在肺腺癌组织中表达升高(P=0.032)。双荧光素酶报告基因发现circUBAP2L序列上位点(CTACCTC-CGACAACCTATACCT)与hsa-let-7c结合,代谢组学(LC-MS)hsa-let-7c启动子序列上GAD1基因位点(AACAUGU)与GAD1结合。转录组学和代谢组学分析结果显示,肺腺癌细胞GAD1过表达增加1-甲基组氨酸(log2FC=2.37,P=0.00023)、L-赤式-4-羟基谷氨酸(log2FC=2.02,P=0.00055)、二甲基甘氨酸(log2FC=2.45,P=0.00129)水平,影响甘氨酸、丝氨酸和苏氨酸代谢通路(P=0.00059)及癌症通路(P=0.00541)。结论 circUBAP2L通过hsa-let-7c/GAD1轴调控肺腺癌细胞中1-甲基组氨酸和二甲基甘氨酸代谢。

Abstract:

Objective To investigate the roles of circular ubiquitin-associated protein 2-like(circUBAP2L), hsa-let-7c, and its target gene glutamate decarboxylase 1(GAD1)in lung adenocarcinoma. Methods Small-RNA sequencing was used to identify differential expression of hsa-let-7c. Bioinformatics analysis and real-time polymerase chain reaction(PCR)were used to assess differential expression of circUBAP2L(the upstream gene of hsa-let-7c)and GAD1(the downstream target gene). Dual-luciferase reporter assays were conducted to validate the regulatory relationships between circUBAP2L and hsa-let-7c, as well as hsa-let-7c and GAD1. A lentivirus was used to construct a GAD1 overexpression model in A549 lung cancer cells. Transcriptomic RNA sequencing(RNA-seq)and metabolomic liquid chromatography-mass spectrometry(LC-MS)were performed to analyze GAD1-associated differential genes and metabolites. Multi-omics integration was applied to explore the impact of GAD1 on amino acid metabolism in lung adenocarcinoma cells. Results Real-time PCR confirmed upregulated expression of circUBAP2L in lung adenocarcinoma(P<0.01). Analysis of TCGA and GTEx databases revealed reduced expression of hsa-let-7c in lung adenocarcinoma tissues(P<0.0001), which was further validated by small-RNA sequencing(P<0.01). Transcriptomic sequencing showed elevated GAD1 expression in lung adenocarcinoma compared to normal tissues(P=0.032). Dual-luciferase reporter assays identified a binding site(CTACCTC-CGACAACCTATACCT)between circUBAP2L and hsa-let-7c, while LC-MS metabolomics revealed that the GAD1 gene locus(AACAUGU)on the hsa-let-7c promoter binds to GAD1. Transcriptomic and metabolomic analyses demonstrated that GAD1 overexpression in lung adenocarcinoma cells increased levels of 1-methylhistidine(log2FC=2.37, P=0.00023), L-erythro-4-hydroxyglutamate(log2FC=2.02, P=0.00055), and dimethylglycine(log2FC=2.45, P=0.00129), affecting glycine, serine, and threonine metabolism pathways(P=0.00059)and cancer-related pathways(P=0.00541). Conclusion circUBAP2L regulates 1-methylhistidine and dimethylglycine metabolism in lung adenocarcinoma cells through the hsa-let-7c/GAD1 axis.

参考文献/References:

1 吴国明, 钱桂生. 非小细胞肺癌靶向治疗研究进展及新理念[J/CD]. 中华肺部疾病杂志(电子版), 2019, 12(4): 405-408.
2 Siegel RL, Miller KD, Jemal A. Cancer statistics[J]. CA Cancer J Clin, 2018, 68(1): 7-30.
3 Cao M, Li H, Sun D, et al. Cancer burden of major cancers in China: A need for sustainable actions[J]. Cancer Commun(Lond), 2020, 40(5): 205-210.
4 Jurisic V, Vukovic V, Obradovic J, et al. EGFR Polymorphism and survival of NSCLC patients treated with TKIs: a systematic review and meta-analysis[J]. J Oncol, 2020, 2020: 1973241.
5 Li J, Zhen L, Zhang Y, et al. Circ-104916 is downregulated in gastric cancer and suppresses migration and invasion of gastric cancer cells[J]. Onco Targets Ther, 2017, 10: 3521-3529.
6 Jeck WR, Sorrentino JA, Wang K, et al. Circular RNAs are abundant, conserved, and associated with ALU repeats[J]. Rna, 2013, 19(2): 141-157.
7 Su H, Tao T, Yang Z, et al. Circular RNA cTFRC acts as the sponge of MicroRNA-107 to promote bladder carcinoma progression[J]. Mol Cancer, 2019, 18(1): 27.
8 Rong D, Lu C, Zhang B, et al. CircPSMC3 suppresses the proliferation and metastasis of gastric cancer by acting as a competitive endogenous RNA through sponging miR-296-5p[J]. Mol Cancer, 2019, 18(1): 25.
9 Xu L, Feng X, Hao X, et al. CircSETD3(Hsa_circ_0000567)acts as a sponge for microRNA-421 inhibiting hepatocellular carcinoma growth[J]. J Exp Clin Cancer Res, 2019, 38(1): 98.
10 Feng J, Chen K, Dong X, et al. Genome-wide identification of cancer-specific alternative splicing in circRNA[J]. Mol Cancer, 2019, 18(1): 35.
11 Subramanian A, Kuehn H, Gould J, et al. GSEA-P: a desktop application for Gene Set Enrichment Analysis[J].Bioinformatics, 2007, 23(23): 3251-3253.
12 Zhao B, Han H, Chen J, et al. MicroRNA let-7c inhibits migration and invasion of human non-small cell lung cancer by targeting ITGB3 and MAP4K3[J]. Cancer Lett, 2014, 342(1): 43-51.
13 Dou Z, Li M, Shen Z, et al. GAD1-mediated GABA elicits aggressive characteristics of human oral cancer cells[J]. Biochem Biophys Res Commun, 2023, 681: 80-89.
14 Tsuboi M, Kondo K, Masuda K, et al. Prognostic significance of GAD1 overexpression in patients with resected lung adenocarcinoma[J]. Cancer Med, 2019, 8(9): 4189-4199.
15 Withanage MHH, Liang H, Zeng E. RNA-seq experiment and data analysis[J]. Methods Mol Biol, 2022, 2418: 405-424.
16 徐 瑜, 白 莉. 广泛期小细胞肺癌免疫治疗新理念[J/CD]. 中华肺部疾病杂志(电子版), 2021, 14(4): 407-411.
17 王洪武, 金发光. 晚期非小细胞肺癌多域整合治疗策略[J/CD]. 中华肺部疾病杂志(电子版), 2022, 15(4): 457-461.
18 Bauermeister A, Mannochio-Russo H, Costa-Lotufo L.V, et al. Mass spectrometry-based metabolomics in microbiome investigations[J]. Nat Rev Microbiol, 2022, 20(3): 143-160.
19 Nie M, Yao K, Zhu X, et al. Evolutionary metabolic landscape from preneoplasia to invasive lung adenocarcinoma[J]. Nat Commun, 2021, 12(1): 6479.
20 Sun X, Nong M, Meng F, et al. Architecting the metabolic reprogramming survival risk framework in LUAD through single-cell landscape analysis: three-stage ensemble learning with genetic algorithm optimization[J]. J Transl Med, 2024, 22(1): 353.
21 Herlihy AE, Boeing S, Weems JC, et al. UBAP2/UBAP2L regulate UV-induced ubiquitylation of RNA polymerase Ⅱ and are the human orthologues of yeast Def1[J]. DNA Repair(Amst), 2022, 115: 103343.
22 Guerber L, Pangou E, Sumara I. Ubiquitin Binding Protein 2-Like(UBAP2L): is it so NICE After All?[J]. Front Cell Dev Biol, 2022, 10: 931115.
23 He J, Chen Y, Cai L, et al. UBAP2L silencing inhibits cell proliferation and G2/M phase transition in breast cancer[J]. Breast Cancer, 2018, 25(2): 224-232.
24 Hansen TB, Venø MT, Damgaard CK, et al. Comparison of circular RNA prediction tools[J]. Nucleic Acids Res, 2016, 44(6): e58.
25 Huang D, Wang Y, Thompson JW, et al. Cancer-cell-derived GABA promotes β-catenin-mediated tumour growth and immunosuppression[J]. Nat Cell Biol, 2022, 24(2): 230-241.
26 Dong Y, Wang G, Nie D, et al. Tumor-derived GABA promotes lung cancer progression by influencing TAMs polarization and neovascularization[J]. Int Immunopharmacol, 2024, 126: 111217.
27 Li Z., Zhang H. Reprogramming of glucose, fatty acid and amino acid metabolism for cancer progression[J]. Cell Mol Life Sci, 2016, 73(2): 377-392.
28 Xu M, Chen J, Peng C, et al. Non-targeted metabolomics analysis of indoleamine 2,3-dioxygenase inhibitor treatment in a mouse model of early-stage lung adenocarcinoma[J]. Transl Cancer Res, 2024, 13(2): 900-915.
29 Gao H, Lu Q, Liu X, et al. Application of 1H NMR-based metabonomics in the study of metabolic profiling of human hepatocellular carcinoma and liver cirrhosis[J]. Cancer Sci, 2009, 100(4): 782-785.
30 Yu M, Wen W, Wang Y, et al. Plasma metabolomics reveals risk factors for lung adenocarcinoma[J]. Front Oncol, 2024, 14: 1277206.
31 Heger Z, Gumulec J, Cernei N, et al. Relation of exposure to amino acids involved in sarcosine metabolic pathway on behavior of non-tumor and malignant prostatic cell lines[J]. Prostate, 2016, 76(7): 679-90.
32 Guo K, Cao Y, Li Z, et al. Glycine metabolomic changes induced by anticancer agents in A549 cells[J]. Amino Acids, 2020, 52(5): 793-809.

备注/Memo

备注/Memo:

基金项目: 2021重庆市自然科学基金面上项目(cstc2021jcyj-msxmX0996)
2023重庆市万州区科卫联合医学科研项目(wzstc-kw2023010)
通信作者: 陈 田, Email: c13883643151@126.com

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更新日期/Last Update: 2025-04-25