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[1]翁晓芹,秦嘉阳,张 扬,等.HIF-1α/VEGF-VEGFR2/Nrp-1表达对NSCLC患者Treg增殖的影响[J].中华肺部疾病杂志,2021,(06):717-722.[doi:10.3877/cma.j.issn.1674-6902.2021.06.003]
　Weng Xiaoqin,Qin Jiayang,Zhang Yang,et al.Effect of HIF-1α/VEGF-VEGFR2/Nrp-1 on Treg proliferation in NSCLC patients[J].,2021,(06):717-722.[doi:10.3877/cma.j.issn.1674-6902.2021.06.003]
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HIF-1α/VEGF-VEGFR2/Nrp-1表达对NSCLC患者Treg增殖的影响(PDF)

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

卷:
期数:
2021年06期
页码:
717-722
栏目:
论著
出版日期:
2021-12-20

文章信息/Info

Title:
Effect of HIF-1α/VEGF-VEGFR2/Nrp-1 on Treg proliferation in NSCLC patients
作者:
翁晓芹; 秦嘉阳; 张 扬; 崔 进; 沈 红
210000 南京,南京医科大学第二附属医院呼吸内科
Author(s):
Weng Xiaoqin;  Qin Jiayang;  Zhang Yang;  Cui Jin;  Shen Hong.
Department of Respiratory, the Second Affiliated Hospital of Nanjing Medical University, Nanjing 210000,China
关键词:
非小细胞肺癌;  调节性T细胞;  缺氧诱导因子-1α;  血管内皮生长因子;  神经纤毛蛋白-1
Keywords:
Non-small cell lung cancer;  Regulatory T Cells;  Hypoxia inducible factor-1α;  Vascular endothelial growth factor;  Neuropilin-1
分类号:
R734.2
DOI:
10.3877/cma.j.issn.1674-6902.2021.06.003
摘要:
目的 分析缺氧诱导因子-1α(HIF-1α)、血管内皮生长因子(VEGF)、血管内皮生长因子受体2(VEGFR2)、神经纤毛蛋白-1(Nrp-1)表达对非小细胞肺癌(NSCLC)患者调节性T细胞( regulatory T Cells, Treg)增殖的影响。方法 纳入60例NSCLC患者及20名健康人群,提取外周血单个核细胞(PBMC),采用流式细胞学检测核内CD4+CD25+FOXP3+Treg细胞含量。使用酶联免疫吸附试验(ELISA)法检测PBMC细胞上清液中HIF-1α、VEGF、转化生长因子-β(TGF-β)、白细胞介素-2(IL-2)、白细胞介素-10(IL-10)的表达水平。Western blotting方法检测Treg细胞上清液中VEGFR2、Nrp-1蛋白表达水平,Transwell迁移实验检测VEGF对Treg细胞的趋化作用。结果 NSCLC患者外周血中CD4+CD25+FOXP3+Treg 细胞比例明显高于健康对照组(P<0.01); 与对照相比,NSCLC组患者HIF-1α、VEGF、IL-10、TGF-β的表达水平明显升高(P<0.01),IL-2的表达水平明显降低(P<0.01); Person相关性分析提示NSCLC患者PBMC中Treg含量与PBMC细胞上清液中HIF-1α、IL-10、TGF-β、VEGF表达均呈正相关(r=0.74; r=0.73; r=0.68; r=0.58,P均<0.01),与IL-2表达呈负相关(r=-0.59,P<0.01)。NSCLC患者Treg细胞表面VEGFR2、Nrp-1受体表达增加(P<0.01); VEGF对Treg细胞有趋化作用,缺氧状态下趋化作用更为强(P<0.01)。结论 NSCLC微环境缺氧,HIF-1α表达上调,肿瘤细胞和内皮细胞分泌VEGF增多,可通过招募和细胞转化促进Treg增多,其表面受体VEGFR2/Nrp-1表达上升,从而Treg细胞积聚活化,促进IL-2、IL-10、TGF-β 细胞因子分泌,进而抑制效应细胞功能,肺癌细胞免疫逃逸,病情进展。
Abstract:
Objective To investigate the effects of hypoxia inducible factor-1α(HIF-1α), vascular endothelial growth factor(VEGF), vascular endothelial growth factor receptor 2(VEGFR2)、neuropilin -1(Nrp-1)expression on the proliferation of regulatory T Cells(Treg)in patients with non-small cell lung cancer(NSCLC). Methods All of 60 non-small cell patients(NSCLC)and 20 healthy people were enrolled and peripheral blood mononuclear cell(PBMC)were extracted. The content of CD4+CD25+FOXP3+Treg cells in PBMC was detected by flow cytometry. The expression levels of HIF-1α, VEGF, transforming growth factor-β(TGF-β), Interleukin-2(IL-2)and interleukin-10(IL-10)in the supernatant of PBMC cells were detected by ELISA. The expression levels of VEGFR2 and Nrp-1 proteins in the supernatant of Treg cells were detected by Westernblotting, and the chemotaxis of VEGF to Treg cells was detected by transwell migration assay. Results The proportion of CD4+CD25+FOXP3+Treg cells in peripheral blood of patients with NSCLC was significantly higher than that of healthy control group(P<0.01). Compared with the normal group, the expression levels of HIF-1α, IL-10, TGF-β and VEGF in the NSCLC group were significantly increased(P<0.01), and the expression level of IL-2 was significantly decreased(P<0.01). The analysis of person correlation suggested that the content of Treg in PBMCs of patients with NSCLC was positively correlated with the expressions of HIF-1α, IL-10, TGF-β, and VEGF in the supernatant of PBMCs(r=0.74; r=0.73; r=0.68; r=0.58, all P<0.01), and negatively correlated with IL-2 expression(r=-0.59, P<0.01). The expression levels of VEGFR2 and Nrp-1 on the surface of Treg cells in patients with NSCLC were increased(P<0.01). VEGF showed a chemotactic effect on Treg cells, especially in the hypoxic state(P<0.01). Conclusion The hypoxia in NSCLC microenvironment induces the up-regulation of HIF-1α expression and the increased secretion of VEGF by tumor cells and endothelial cells. The recruitment and cell transformation promote the increase of Treg and the expression of surface receptor VEGFR2/Nrp-1, which leads to the accumulation and activation of Treg cells, Treg cells can stimulate the secretion of IL-2, IL-10, TGF-β, and then inhibit the effector cell function, leading to immune escape of lung cancer cells and progression of the disease.

参考文献/References:

1 Bade BC, Dela Cruz CS. Lung Cancer 2020: Epidemiology, etiology, and prevention[J]. Clin Chest Med, 2020, 41(1): 1-24.
2 Senbabaoglu Y, Gejman RS, Winer AG, et al. Tumor immune microenvironment characterization in clear cell renal cell carcinoma identifies prognostic and immunotherapeutically relevant messenger RNA signatures [J]. Genome Biol, 2016, 17(1): 231.
3 胡晓雨, 刘 超, 岳金波. Ⅳ期非小细胞肺癌患者外周血Treg细胞与预后相关性分析[J]. 中华肿瘤防治杂志, 2020, 27(9): 720-724,729.
4 马维江, 王 平, 杨立民, 等. 辅助性T细胞17和调节性T细胞的免疫功能及其对非小细胞肺癌免疫调节作用的研究进展[J]. 肿瘤研究与临床, 2020,(1): 66-69.
5 马建梅, 陈 承, 张星星, 等. PEDF下调HIF-1α表达抑制缺氧状态下NSCLC细胞增殖、迁移的作用机制[J]. 现代肿瘤医学, 2019, 27(17): 3005-3009.
6 He LY, Li L, Guo ML, et al. Relationship between CD4+CD25+Treg and expression of HIF-1αand Ki-67 in NSCLC patients[J]. Eur Rev Med Pharmacol Sci, 2015, 19(8): 1351-1355.
7 Long Y, Tao H, Karachi A, et al. Dysregulation of glutamate transport enhances treg function that promotes VEGF blockade resistance in glioblastoma[J]. Cancer Res, 2020, 80(3): 499-509.
8 Voron T, Colussi O, Marcheteau E, et al. VEGF-A modulates expression of inhibitory checkpoints on CD8+ T cells in tumors[J]. J Exp Med, 2015, 212(2): 139-148.
9 Suzuki H, Onishi H, Wada J, et al. VEGFR2 is selectively expressed by FOXP3high CD4+ Treg[J]. Eur J Immunol, 2010, 40(1): 197-203.
10 Leclerc M, Voilin E, Gros G, et al. Regulation of antitumour CD8 T-cell immunity and checkpoint blockade immunotherapy by Neuropilin-1[J]. Nat Commun, 2019, 10(1): 3345.
11 Otsuki Y, Saya H, Arima Y. Prospects for new lung cancer treatments that target EMT signaling[J]. Dev Dyn, 2018, 247(3): 462-472.
12 Valcourt U, Carthy J, Okita Y, et al. Analysis of epithelialmesenchymal transition induced by transforming growth factor beta[J]. Methods Mol Biol, 2016, 1344: 147-181.
13 钱桂生. 肺癌不同病理类型发病率的变化情况及其原因[J/CD]. 中华肺部疾病杂志(电子版), 2011, 4(1): 1-5.
14 Noman MZ, Hasmim M, Messai Y, et al. Hypoxia: a key player in antitumor immune response. A Review in the Theme: Cellular Responses to Hypoxia[J]. Am J Physiol Cell Physiol, 2015, 309(9): C569-C579.
15 Balamurugan K. HIF-1 at the crossroads of hypoxia, inflammation, and cancer[J]. Int J Cancer, 2016, 138(5): 1058-1066.
16 Apte RS, Chen DS, Ferrara N. VEGF in signaling and disease: Beyond discovery and development[J]. Cell, 2019, 176(6): 1248-1264.
17 Yang J, Yan J, Liu B. Targeting VEGF/VEGFR to modulate antitumor immunity[J]. Front Immunol, 2018, 9: 978.
18 Yang J, Yan J, Shao J, et al. Immune-mediated antitumor effect by VEGFR2 selective inhibitor for gastric cancer[J]. Onco Targets Ther, 2019, 12: 9757-9765.
19 Hu C, Jiang X. Role of NRP-1 in VEGF-VEGFR2-independent tumorigenesis[J]. Target Oncol, 2016, 11(4): 501-505.
20 卜晓红. VEGFR2、NRP-1在非小细胞肺癌中的表达及意义分析[J]. 中国现代药物应用, 2016, 10(20): 36-37.
21 施星月, 陈 翀, 刘峰林, 等. Neuropilin-1在非小细胞肺癌患者外周血Treg、PDC细胞中的表达及临床意义[J]. 肿瘤防治研究, 2020, 47(7): 531-535.
22 王洪建, 刘小慧, 刘燕飞, 等. Th1与Th2细胞因子及血管内皮生长因子联合检测对肺癌患者发生发展及临床诊断价值分析[J]. 实验与检验医学, 2019, 37(6): 1063-1066.
23 李晓英, 谢启超. 不同分期非小细胞肺癌与Treg细胞、细胞因子相关研究[J]. 安徽医科大学学报, 2017, 52(1): 113-116.
24 Yan X, Jiao SC, Zhang GQ, et al. Tumor-associated immune factors are associated with recurrence and metastasis in non-small cell lung cancer[J]. Cancer Gene Ther, 2017, 24(2): 57-63.
25 Xu Y, Pan Y, Zhou Z. Recombinant human thrombopoietin combined with interleukin-2 improves the effects of chemosensitivity and thrombocytopenia on a basic gemcitabine and carboplatin combination therapy for non-small cell lung cancer in a nude mouse model[J]. J Thorac Dis, 2019, 11(11): 4671-4681.
26 Ohue Y, Nishikawa H. Regulatory T(Treg)cells in cancer: Can treg cells be a new therapeutic target?[J]. Cancer Sci, 2019, 110(7): 2080-2089.
27 Zhao Y, Chen S, Shen F, et al. In vitro neutralization of autocrine IL10 affects Op18/stathmin signaling in nonsmall cell lung cancer cells[J]. Oncol Rep, 2019, 41(1): 501-511.
28 Yang L, Dong Y, Li Y, et al. IL-10 derived from M2 macrophage promotes cancer stemness via JAK1/STAT1/NF-κB/Notch1 pathway in non-small cell lung cancer[J]. Int J Cancer, 2019, 145(4): 1099-1110.
29 刘瑞敏, 张爱红, 晁玮霞, 等. 高表达Foxp3的肺癌细胞抑制人CD4+ T细胞免疫活性[J]. 中国免疫学杂志, 2017, 33(8): 1141-1145.
30 梁 晶. 肺癌患者外周血Treg细胞与IL-10、TGF-β检测及其临床意义[J]. 临床肺科杂志, 2015, 20(11): 1980-1983.
31 Oh E, Hong J, Yun CO. Regulatory T Cells induce metastasis by increasing TGF-β and enhancing the epithelial-mesenchymal transition[J]. Cells, 2019, 8(11): 1387.
32 Wang D, Yang L, Yue D, et al. Macrophage-derived CCL22 promotes an immunosuppressive tumor microenvironment via IL-8 in malignant pleural effusion[J]. Cancer Lett, 2019, 452: 244-253.

备注/Memo

备注/Memo:
基金项目: 南京市医学科技发展资金(YKK16226)
通信作者: 沈 红, Email: shenhong603@163.com
更新日期/Last Update: 2021-12-20