切换至 "中华医学电子期刊资源库"

中华肺部疾病杂志(电子版) ›› 2023, Vol. 16 ›› Issue (01) : 31 -38. doi: 10.3877/cma.j.issn.1674-6902.2023.01.007

论著

EGLN3在肺腺癌中表达的Meta分析
杨硕1, 马洪明2, 关晓婷1, 陈正贤3,()   
  1. 1. 511300 广州,前海人寿广州总医院呼吸与危重症医学科
    2. 510630 广州,暨南大学附属第一医院呼吸与危重症医学科
    3. 511300 广州,前海人寿广州总医院呼吸与危重症医学科;510665 广州,中山大学附属第六医院呼吸与危重症医学科
  • 收稿日期:2022-11-09 出版日期:2023-02-25
  • 通信作者: 陈正贤
  • 基金资助:
    天河区科技研究资助项目(201404KW010)

Meta-analysis of EGLN3 expression in lung adenocarcinoma

Shuo Yang1, Hongming Ma2, Xiaoting Guan1, Zhengxian Chen3,()   

  1. 1. Foresea Life Insurance Guangzhou General Hospital Pulmonary and Critical Care Medicine, Guangzhou 511300, China
    2. The First Affiliated Hospital of Jinan University Pulmonary and Critical Care Medicine, Guangzhou 511300, China
    3. Foresea Life Insurance Guangzhou General Hospital Pulmonary and Critical Care Medicine, Guangzhou 511300, China; The sixth affiliated Hospital of Sun Yat-sen University Pulmonary and Critical Care Medicine, Guangzhou 510665, China
  • Received:2022-11-09 Published:2023-02-25
  • Corresponding author: Zhengxian Chen
引用本文:

杨硕, 马洪明, 关晓婷, 陈正贤. EGLN3在肺腺癌中表达的Meta分析[J]. 中华肺部疾病杂志(电子版), 2023, 16(01): 31-38.

Shuo Yang, Hongming Ma, Xiaoting Guan, Zhengxian Chen. Meta-analysis of EGLN3 expression in lung adenocarcinoma[J]. Chinese Journal of Lung Diseases(Electronic Edition), 2023, 16(01): 31-38.

目的

分析EGLN3在肺腺癌(lung adenocarcinoma, LUAD)中的作用。

方法

(1)通过癌症基因组图谱(the cancer genome atlas, TCGA)数据库下载535例肺腺癌样本和59例正常样本,分析EGLN3在肺腺癌和正常肺组织中的表达差异;(2)使用R语言分析EGLN3的表达水平及其在肺腺癌中的临床意义;(3)通过Kaplan-Meier方法、单因素和多因素COX回归分析和生存预测列线图确定EGLN3的表达水平与肺腺癌预后的关系;(4)通过基因集富集分析(GSEA)筛选EGLN3相关的生物学途径;(5)细胞实验:通过蛋白质印迹法(WB)验证EGLN3在肺腺癌细胞中是否存在差异表达;对EGLN3基因进行过表达和干扰表达,采用CCK-8增殖实验、划痕实验和Transwell侵袭实验,明确表达水平对肺腺癌细胞增殖、迁移和侵袭的影响。

结果

(1)EGLN3在肺腺癌组织中表达水平高于正常肺组织细胞;(2)EGLN3表达水平与肺腺癌病理分期相关,病理分期越高,对应EGLN3表达水平越高;(3)EGLN3表达水平与肺腺癌总生存期(OS)独立预后相关,EGLN3高表达肺腺癌较EGLN3低表达肺腺癌OS、疾病特异性生存期(DSS)和无进展间隔(PFI)差;(4)基因集富集分析预测EGLN3可能参与的信号通路包括激活MYC基因、上皮细胞-间充质转化(EMT)、DNA复制、细胞周期、mTORC1通路、G2/M检查点、E2F转录过程、P53信号通路等;(5)细胞实验:EGLN3在肺腺癌细胞中表达水平高于人支气管上皮样细胞;CCK-8增殖实验、划痕实验和Transwell侵袭实验显示,EGLN3过表达促进肺腺癌细胞增殖、侵袭和迁移,EGLN3干扰表达抑制肺腺癌细胞增殖、侵袭和迁移。

结论

EGLN3在肺腺癌细胞中存在高表达,提示肺腺癌预后不良,可作为肺腺癌预后的预测因素。EGLN3表达可促进肺腺癌细胞增殖、侵袭和迁移。

Objective

EGLN3 is differentially expressed in many kinds of tumors. However, the role of EGLN3 in lung adenocarcinoma (LUAD) is not clear. The purpose of this study is to explore the role of EGLN3 in lung adenocarcinoma.

Methods

(1)535 LUAD samples and 59 normal samples were downloaded from Cancer Genome Map (TCGA) database to analyze the difference of EGLN3 expression between LUAD and normal lung tissues. (2)The expression level of EGLN3 and its clinical significance in LUAD were analyzed by R language. (3)The relationship between the expression of EGLN3 and the prognosis of LUAD was determined by Kaplan-Meier method, univariate and multivariate COX regression analysis and survival prediction diagram. (4)The biological pathways related to EGLN3 were screened by gene sets enrichment analysis (GSEA). (5)Cell experiment: Western blotting (WB) was used to verify the differential expression of EGLN3 in lung adenocarcinoma cells, and then EGLN3 was overexpressed and interfered, and CCK-8 proliferation test, scratch test and Transwell invasion test were carried out to determine the effect of its expression level on the proliferation, migration and invasion of lung adenocarcinoma cells.

Results

The expression level of EGLN3 in LUAD tissue was significantly higher than that in normal lung tissue. (2)The expression level of EGLN3 is related to the pathological stage of lung adenocarcinoma. The higher the pathological stage, the higher the expression level of EGLN3. (3)The expression of EGLN3 is an independent prognostic factor related to the overall survival (OS) of lung adenocarcinoma. The patients with high expression of EGLN3 had worse OS, disease specific survival (DSS) and progress free interval (PFI) than those with low expression of EGLN3. (4)Gene sets enrichment analysis predicted the possible signal pathways involved in EGLN3, including activation of MYC gene, epithelial mesenchymal transformation (EMT), DNA replication, cell cycle, mTORC1 pathway, G2/M checkpoint, E2F transcription process, p53 signal pathway, etc. (5) Cell experiment: The expression level of EGLN3 in LUAD cells was significantly higher than that in human bronchial epithelioid cells. CCK-8 proliferation test, scratch test and Transwell invasion test: the overexpression of EGLN3 promoted the proliferation, invasion and migration of lung adenocarcinoma cells, while the interference expression of EGLN3 inhibited the proliferation, invasion and migration of lung adenocarcinoma cells.

Conclusions

EGLN3 is highly expressed in lung adenocarcinoma cells, and its high expression indicates a poor prognosis of lung adenocarcinoma, which can be used as an independent factor to predict the prognosis of lung adenocarcinoma. The expression of EGLN3 can promote the proliferation, invasion and migration of lung adenocarcinoma cells.

表1 EGLN3表达水平与肺腺癌临床特征的相关性[n(%)]
图1 基于TCGA数据库EGLN3基因mRNA表达水平。注:A:肺腺癌组织和正常组织间EGLN3表达水平;B:59个匹配的肺腺癌组织和相应正常组织中表达水平
图2 EGLN3 mRNA表达与肺腺癌临床特征关系。注:A:临床分期;B:T分期;C:N分期;D:M分期;E:吸烟时间;F:初次治疗结果
图3 EGLN3高表达、低表达的总生存率、疾病特异性生存率和无进展间隔率。注:A:总生存期(OS);B:疾病特异性生存期(DSS);C:无进展间隔(PFI);D:接受者操作特征曲线(ROC)
图4 EGLN3在预测肺腺癌患者预后中的作用;注:A:生存概率预测的列线图;B:校准曲线
图5 EGLN3在肺腺癌细胞中高表达
图6 EGLN3基因过表达后PC9细胞增殖、侵袭和迁移能力增强。注:A:CCK-8增殖实验:EGLN3基因过表达组PC9细胞增殖能力大于对照组;B:划痕实验:EGLN3基因过表达组PC9细胞迁移能力大于对照组;C:Transwell侵袭实验:EGLN3基因过表达组PC9细胞侵袭能力大于对照组
图7 EGLN3基因干扰表达后SPCA-1细胞增殖、侵袭和迁移能力减弱。注:A:CCK-8增殖实验:EGLN3基因干扰表达后SPCA -1细胞增殖能力低于对照组;B:划痕实验:EGLN3基因干扰表达后SPCA -1细胞迁移能力低于对照组;C:Transwell侵袭实验:EGLN3基因干扰表达后SPCA -1细胞侵袭能力低于对照组
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, 2020[J]. CA: A Cancer J Clin, 2020, 70(1): 7-30.
3
Jiang F, Liang M, Huang X, et al. High expression of PIMREG predicts poor survival outcomes and is correlated with immune infiltrates in lung adenocarcinoma[J]. Peer J, 2017, 9: e11697.
4
陈艳丽,王媛媛,张 勇,等. 中晚期非小细胞肺癌患者化疗前后T淋巴细胞亚群表达差异分析及临床意义[J/CD]. 中华肺部疾病杂志(电子版), 2020, 13(1): 13-17.
5
中华医学会肿瘤学分会,中华医学会杂志社. 中华医学会肿瘤学分会肺癌临床诊疗指南(2021版)[J]. 中华肿瘤杂志2021, 43(6): 591-621.
6
Torre LA, Siegel RL, Jemal A. Lung cancer statistics.[J]. Adv Exper Med Biol, 2016, 893: 1-19.
7
Yue Y, Cui J, Zhao Y, et al. Circ_101341 deteriorates the progression of clear cell renal cell carcinoma through the miR- 411/EGLN3 axis[J]. Cancer Manag Res, 2020, 12: 13513-13525.
8
Xie L, Xiao K, Whalen EJ, et al. Oxygen-regulated beta(2)-adrenergic receptor hydroxylation by EGLN3 and ubiquitylation by pVHL[J]. Sci Signal, 2009, 2(78): ra33.
9
Luo W, Hu H, Chang R, et al. Pyruvate kinase M2 is a PHD3-stimulated coactivator for hypoxia-inducible factor 1[J]. Cell, 2011, 145(5): 732-744.
10
Chen N, Rinner O, Czernik D, et al. The oxygen sensor PHD3 limits glycolysis under hypoxia via direct binding to pyruvate kinase[J]. Cell Res, 2011, 21(6): 983-986.
11
Xie L, Pi X, Mishra A, et al. PHD3-dependent hydroxylation of HCLK2 promotes the DNA damage response[J]. J Clin Invest, 2012122(8): 2827-2836.
12
Yan B, Jiao S, Zhang HS, et al. Prolyl hydroxylase domain protein 3 targets Pax2 for destruction[J]. Biochem Biophys Res Commun, 2011, 409(2): 315-320.
13
Su Y, Loos M, Giese N, et al. PHD3 regulates differentiation, tumour growth and angiogenesis in pancreatic cancer[J]. Br J Cancer, 2010, 103(10): 1571-1579.
14
Schlisio S, Kenchappa RS, Vredeveld L, et al. The kinesin KIF1Bβ acts downstream from EglN3 to induce apoptosis and is a potential 1p36 tumor suppressor[J]. Genes Development, 2008, 22(7): 884-893.
15
Jr. WGK, Bommi-Reddy A, Schlisio S. Inhibitors of egln3 activity for the treatment of neurodegenerative disorders: EP, EP1912637 A2[P]. 2010.
16
Li S, Rodriguez J, Li W, et al. EglN3 hydroxylase stabilizes BIM-EL linking VHL type 2C mutations to pheochromocytoma pathogenesis and chemotherapy resistance[J]. Proc Natl Acad Sci U S A, 2019, 116(34): 16997-17006.
17
Wang Y, Li X, Liu W, et al. MicroRNA-1205, encoded on chromosome 8q24, targets EGLN3 to induce cell growth and contributes to risk of castration-resistant prostate cancer[J]. Oncogene, 2019, 38(24): 4820-4834.
18
Pescador N, Cuevas Y, Naranjo S, et al. Identification of a functional hypoxia-responsive element that regulates the expression of the egl nine homologue 3 (egln3/phd3) gene[J]. Biochem J, 2005, 390(1): 189-197.
19
蔡凤琳,邓靖宇. EGLN3在肿瘤发生发展中作用的研究进展[J]. 天津医科大学学报2020, 26(6): 5.
20
Kim KH, Lee HH, Yoon YE, et al. Prolyl hydroxylase-3 is a novel renal cell carcinoma biomarker[J]. Investig Clin Urol, 2019, 60(6): 425-431.
21
Subramanian A, Tamayo P, Mootha VK, et al. Gene set enrichment analysis: A knowledge-based approach for interpreting genome-wide expression profiles[J]. Proc Natl Acad Sci U S A, 2005, 102(43): 15545-15550.
22
Canzler S, Hackermüller J. multiGSEA: a GSEA-based pathway enrichment analysis for multi-omics data[J]. BMC Bioinformatics, 2020, 21(1): 561.
23
Gatenby RA, Gillies RJ. Why do cancers have high aerobic glycolysis?[J]. Nat Rev Cancer, 2004, 4(11): 891-899.
24
Harris AL. Hypoxia-a key regulatory factor in tumour growth[J]. Nat Rev Cancer, 2002, 2(1): 38-47.
25
Semenza GL. Targeting HIF-1 for cancer therapy[J]. Nat Rev Cancer2003, 3(10): 721-732.
26
Frey L, Klümper N, Schmidt D, et al. CircEHD2, CircNETO2 and CircEGLN3 as diagnostic and prognostic biomarkers for patients with renal cell carcinoma[J]. Cancers, 2021, 13(9): 2177.
27
Melillo G. Targeting hypoxia cell signaling for cancer therapy[J]. Cancer Metastasis Rev, 2007, 26(2): 341-352.
28
Semenza GL. Defining the role of hypoxia-inducible factor 1 in cancer biology and therapeutics[J]. Oncogene, 2010, 29(5): 625-634.
29
Wheaton WW, Chandel NS. Hypoxia.2.Hypoxia regulates cellular metabolism[J]. Am J Physiol Cell Physiol, 2011, 300(3): C385-C393.
30
Epstein AC, Gleadle JM, McNeill LA, et al. C.elegans EGL-9 and mammalian homologs define a family of dioxygenases that regulate HIF by prolyl hydroxylation[J]. Cell, 2001, 107(1): 43-54.
31
Huang J, Zhao Q, Mooney SM, et al. Sequence determinants in hypoxia-inducible factor-1alpha for hydroxylation by the prolyl hydroxylases PHD1,PHD2,and PHD3[J]. J Biol Chem, 2002, 277(42): 39792-39800.
32
Lee S, Nakamura E, Yang H, et al. Neuronal apoptosis linked to EglN3 prolyl hydroxylase and familial pheochromocytoma genes: developmental culling and cancer[J]. Cancer Cell, 2005, 8(2): 155-167.
33
Lin L, Lin L, Cai J. Circular RNA circ-EGLN3 promotes renal cell carcinoma proliferation and aggressiveness via miR-1299-mediated IRF7 activation[J]. J Cell Biochem, 2020, 121(11): 4377-4385.
34
Xia YJ, Jiang XT, Jiang SB, et al. PHD3 affects gastric cancer progression by negatively regulating HIF1A[J]. Mol Med Rep, 2017, 16(5): 6882.
35
Mao K, You C, Lei D, et al. Potential regulation of glioma through the induction of apoptosis signaling via Egl-9 family hypoxia-inducible factor 3[J]. Oncol Lett, 2017, 13(2): 893-897.
36
Li J, Chen C, Li C, et al. Genome-wide knockout screen identifies EGLN3 involving in ammonia neurotoxicity[J]. Front Cell Dev Biol, 2022, 10: 820692.
37
Fu J. Catalytic-independent inhibition of cIAP1-mediated RIP1 ubiquitination by EGLN3[J]. Cell Signal, 2016, 28(2): 72-80.
38
Zhong C, Li S, Li J, et al. Polymorphisms in the Egl nine homolog 3 (EGLN3) and Peroxisome proliferator activated receptor-alpha (PPARα) genes and their correlation with hypoxia adaptation in Tibetan chickens[J]. PLoS One, 2018, 13(3): e0194156.
[1] 张思平, 刘伟, 马鹏程. 全膝关节置换术后下肢轻度内翻对线对疗效的影响[J]. 中华关节外科杂志(电子版), 2023, 17(06): 808-817.
[2] 罗旺林, 杨传军, 许国星, 俞建国, 孙伟东, 颜文娟, 冯志. 开放性楔形胫骨高位截骨术不同植入材料的Meta分析[J]. 中华关节外科杂志(电子版), 2023, 17(06): 818-826.
[3] 马鹏程, 刘伟, 张思平. 股骨髋臼撞击综合征关节镜手术中闭合关节囊的疗效影响[J]. 中华关节外科杂志(电子版), 2023, 17(05): 653-662.
[4] 陈宏兴, 张立军, 张勇, 李虎, 周驰, 凡一诺. 膝骨关节炎关节镜清理术后中药外用疗效的Meta分析[J]. 中华关节外科杂志(电子版), 2023, 17(05): 663-672.
[5] 李越洲, 张孔玺, 李小红, 商中华. 基于生物信息学分析胃癌中PUM的预后意义[J]. 中华普通外科学文献(电子版), 2023, 17(06): 426-432.
[6] 李雄雄, 周灿, 徐婷, 任予, 尚进. 初诊导管原位癌伴微浸润腋窝淋巴结转移率的Meta分析[J]. 中华普通外科学文献(电子版), 2023, 17(06): 466-474.
[7] 张再博, 王冰雨, 焦志凯, 檀碧波. 胃癌术后下肢深静脉血栓危险因素的Meta分析[J]. 中华普通外科学文献(电子版), 2023, 17(06): 475-480.
[8] 武慧铭, 郭仁凯, 李辉宇. 机器人辅助下经自然腔道取标本手术治疗结直肠癌安全性和有效性的Meta分析[J]. 中华普通外科学文献(电子版), 2023, 17(05): 395-400.
[9] 莫闲, 杨闯. 肝硬化患者并发门静脉血栓危险因素的Meta分析[J]. 中华普外科手术学杂志(电子版), 2023, 17(06): 678-683.
[10] 张圣平, 邓琼, 张颖, 张建文, 梁辉, 王铸. 孤儿核受体HNF4α在肾透明细胞癌中的表达及意义[J]. 中华腔镜泌尿外科杂志(电子版), 2023, 17(06): 627-632.
[11] 段文忠, 白延霞, 徐文亭, 祁虹霞, 吕志坚. 七氟烷和丙泊酚在肝切除术中麻醉效果比较Meta分析[J]. 中华肝脏外科手术学电子杂志, 2023, 12(06): 640-645.
[12] 张维志, 刘连新. 基于生物信息学分析IPO7在肝癌中的表达及意义[J]. 中华肝脏外科手术学电子杂志, 2023, 12(06): 694-701.
[13] 陈安, 冯娟, 杨振宇, 杜锡林, 柏强善, 阴继凯, 臧莉, 鲁建国. 基于生物信息学分析CCN4在肝细胞癌中表达及其临床意义[J]. 中华肝脏外科手术学电子杂志, 2023, 12(06): 702-707.
[14] 杨海龙, 邓满军, 樊羿辰, 徐梦钰, 陈芳德, 吴威浩, 张生元. 腹腔镜胆总管探查术一期缝合术后胆漏危险因素Meta分析[J]. 中华肝脏外科手术学电子杂志, 2023, 12(05): 545-550.
[15] 王苏贵, 皇立媛, 姜福金, 吴自余, 张先云, 李强, 严大理. 异质性细胞核核糖蛋白A2B1在前列腺癌中的作用及其靶向中药活性成分筛选研究[J]. 中华临床医师杂志(电子版), 2023, 17(06): 731-736.
阅读次数
全文


摘要