Hypoxic Macrophage-Derived VEGF Promotes Proliferation and Invasion of Gastric Cancer Cells

  • Fei Ma
  • Bin Zhang
  • Sheqing Ji
  • Hongtao Hu
  • Ye Kong
  • Yawei Hua
  • Suxia LuoEmail author
Original Article



Gastric cancer (GC) is one of the most common causes of cancer death. Hypoxia is an important property of the tumor microenvironment of GC. Increasing evidence demonstrates that tumor-associated macrophages are related to the metastasis of GC, while the precise mechanism of how hypoxic macrophages affect tumor progression is still not fully understood.


To examine whether the mediators released from hypoxic macrophages contribute to the invasion and proliferation of GC cells.


Cell Counting Kit-8 was utilized to determine the proliferation of SGC7901 and MKN45 cells. The invasion of SGC7901 and MKN45 cells was measured by transwell invasion assay. Expression of VEGF mRNA in THP-1-derived macrophages was determined by RT-PCR, and protein level of VEGF in the culture medium was detected by ELISA.


The proliferation and invasion of SGC7901 and MKN45 cells were dramatically increased after treatment with conditioned medium (CM) collected from THP-1-derived macrophages under hypoxia (H-CM), and the phosphorylation of Akt and p38 in SGC7901 and MKN45 cells was also up-regulated by H-CM stimulation. Notably, blockage of PI3K-Akt or p38 MAP kinase abolished the effects of H-CM on the proliferation and invasion of SGC7901 and MKN45 cells. Furthermore, VEGF was increased in macrophages after hypoxia and administration with nintedanib, an inhibitor of VEGFR, significantly decreases the phosphorylation of Akt and p38, as well as the proliferation and invasion of SGC7901 and MKN45 cells in response to H-CM.


Our findings suggest that hypoxia-injured macrophages contribute to the proliferation and invasion of GC cells through the release of mediators such as VEGF.


Gastric cancer Hypoxia Macrophage VEGF 



This study was funded by the National Natural Science Foundation of China under Grant No. U1504816.

Author's contribution

FM, BZ, and SJ performed the experiments of this study. SL, HH, and YK participated in its design and coordination and interpretation of results and helped to draft the manuscript. YH and SL wrote the manuscript. All authors read and approved the final manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interests.


  1. 1.
    Suganuma M, Watanabe T, Yamaguchi K, et al. Human gastric cancer development with TNF-alpha-inducing protein secreted from Helicobacter pylori. Cancer Lett. 2012;322:133–138.CrossRefGoogle Scholar
  2. 2.
    Otto C, Kaemmerer U, Illert B, et al. Growth of human gastric cancer cells in nude mice is delayed by a ketogenic diet supplemented with omega-3 fatty acids and medium-chain triglycerides. BMC Cancer. 2008;8:122.CrossRefGoogle Scholar
  3. 3.
    Wessler S, Krisch LM, Elmer DP, Aberger F. From inflammation to gastric cancer—the importance of Hedgehog/GLI signaling in Helicobacter pylori-induced chronic inflammatory and neoplastic diseases. Cell Commun Signal. 2017;15:15.CrossRefGoogle Scholar
  4. 4.
    Cao Y, Bao S, Yang W, et al. Epigallocatechin gallate prevents inflammation by reducing macrophage infiltration and inhibiting tumor necrosis factor-alpha signaling in the pancreas of rats on a high-fat diet. Nutr Res. 2014;34:1066–1074.CrossRefGoogle Scholar
  5. 5.
    Kamate C, Baloul S, Grootenboer S, et al. Inflammation and cancer, the mastocytoma P815 tumor model revisited: triggering of macrophage activation in vivo with pro-tumorigenic consequences. Int J Cancer. 2002;100:571–579.CrossRefGoogle Scholar
  6. 6.
    Noda S, Yashiro M, Nshii T, Hirakawa K. Hypoxia upregulates adhesion ability to peritoneum through a transforming growth factor-beta-dependent mechanism in diffuse-type gastric cancer cells. Eur J Cancer. 2010;46:995–1005.CrossRefGoogle Scholar
  7. 7.
    Liu L, Sun L, Zhao P, et al. Hypoxia promotes metastasis in human gastric cancer by up-regulating the 67-kDa laminin receptor. Cancer Sci. 2010;101:1653–1660.CrossRefGoogle Scholar
  8. 8.
    Shen Z, Kauttu T, Seppanen H, et al. Both macrophages and hypoxia play critical role in regulating invasion of gastric cancer in vitro. Acta Oncol. 2013;52:852–860.CrossRefGoogle Scholar
  9. 9.
    Guleng B, Han J, Yang JQ, et al. TFF3 mediated induction of VEGF via hypoxia in human gastric cancer SGC-7901 cells. Mol Biol Rep. 2012;39:4127–4134.CrossRefGoogle Scholar
  10. 10.
    Bilgic CI, Tez M. Serum VEGF levels in gastric cancer patients: correlation with clinicopathological parameters. Turk J Med Sci. 2015;45:112–117.CrossRefGoogle Scholar
  11. 11.
    Ozdemir F, Akdogan R, Aydin F, et al. The effects of VEGF and VEGFR-2 on survival in patients with gastric cancer. J Exp Clin Cancer Res. 2006;25:83–88.Google Scholar
  12. 12.
    Roukos DH, Liakakos T, Karatzas G, Kappas AM. Can VEGF-D and VEGFR-3 be used as biomarkers for therapeutic decisions in patients with gastric cancer? Nat Clin Pract Oncol. 2006;3:418–419.CrossRefGoogle Scholar
  13. 13.
    Abdel-Rahman O. Targeting vascular endothelial growth factor (VEGF) pathway in gastric cancer: preclinical and clinical aspects. Crit Rev Oncol Hematol. 2015;93:18–27.CrossRefGoogle Scholar
  14. 14.
    Ayremlou N, Mozdarani H, Mowla SJ, Delavari A. Increased levels of serum and tissue miR-107 in human gastric cancer: correlation with tumor hypoxia. Cancer Biomark. 2015;15:851–860.CrossRefGoogle Scholar
  15. 15.
    Grimshaw MJ, Balkwill FR. Inhibition of monocyte and macrophage chemotaxis by hypoxia and inflammation—a potential mechanism. Eur J Immunol. 2001;31:480–489.CrossRefGoogle Scholar
  16. 16.
    Egners A, Erdem M, Cramer T. The response of macrophages and neutrophils to hypoxia in the context of cancer and other inflammatory diseases. Mediators Inflamm. 2016;2016:2053646.CrossRefGoogle Scholar
  17. 17.
    Griffiths L, Binley K, Iqball S, et al. The macrophage - a novel system to deliver gene therapy to pathological hypoxia. Gene Ther. 2000;7:255–262.CrossRefGoogle Scholar
  18. 18.
    Wu MH, Lee WJ, Hua KT, et al. Macrophage infiltration induces gastric cancer invasiveness by activating the beta-catenin pathway. PLoS ONE. 2015;10:e0134122.CrossRefGoogle Scholar
  19. 19.
    Zhang WJ, Chen C, Zhou ZH, et al. Hypoxia-inducible factor-1 alpha correlates with tumor-associated macrophages infiltration, influences survival of gastric cancer patients. J Cancer. 2017;8:1818–1825.CrossRefGoogle Scholar
  20. 20.
    Zhou H, Wu J, Wang T, et al. CXCL10/CXCR20 axis promotes the invasion of gastric cancer via PI3K/AKT pathway-dependent MMPs production. Biomed Pharmacother. 2016;82:479–488.CrossRefGoogle Scholar
  21. 21.
    Riquelme I, Tapia O, Espinoza JA, et al. The gene expression status of the PI3K/AKT/mTOR pathway in gastric cancer tissues and cell lines. Pathol Oncol Res. 2016;22:797–805.CrossRefGoogle Scholar
  22. 22.
    Yan X, Rui X, Zhang K. Baicalein inhibits the invasion of gastric cancer cells by suppressing the activity of the p38 signaling pathway. Oncol Rep. 2015;33:737–743.CrossRefGoogle Scholar
  23. 23.
    Deng W, Zhang Y, Gu L, et al. Heat shock protein 27 downstream of P38-PI3K/Akt signaling antagonizes melatonin-induced apoptosis of SGC-7901 gastric cancer cells. Cancer Cell Int. 2016;16:5.CrossRefGoogle Scholar
  24. 24.
    Yonemura Y, Fushida S, Bando E, et al. Lymphangiogenesis and the vascular endothelial growth factor receptor (VEGFR)-3 in gastric cancer. Eur J Cancer. 2001;37:918–923.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of General SurgeryAffiliated Cancer Hospital of Zhengzhou UniversityZhengzhouChina
  2. 2.Department of Intervention RadiologyAffiliated Cancer Hospital of Zhengzhou UniversityZhengzhouChina
  3. 3.Department of Medical OncologyAffiliated Cancer Hospital of Zhengzhou UniversityZhengzhouChina

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