Skip to main content
SpringerLink
Log in

TFF3 mediated induction of VEGF via hypoxia in human gastric cancer SGC-7901 cells

  • Published:
Molecular Biology Reports Aims and scope Submit manuscript

Abstract

Increasing evidence indicates that in gastric epithelial cells, induction of TFF3 by hypoxia is mediated by HIF-1. Since VEGF is one of the most important angiogenic factors on cancer progression, we have started to investigate the possible link among HIF-1α, VEGF, and TFF3 in gastric cancer cells. We induced the hypoxic condition in SGC-7901cells using hypoxia-mimetic agent of CoCI2. SGC7901 cells were transfected with pcPUR + U6 plasmid carrying RNAi targeted to human TFF3 and selected puromycin-resistant pools to establish the stable knockdown of TFF3 cells. Our results showed the induction of HIF-1a via hypoxia and consequences of increased expressions of the TFF3 and VEGF in gastric cancer SGC-7901 cells. Overexpression of TFF3 upregulated the mRNA expressions of VEGF and HIF-1a induced by hypoxia, and stable knockdown of TFF3 impaired the mRNA upregulations of VEGF and HIF-1a induced by hypoxia. Furthermore, knockdown of TFF3 reduced the VEGF protein secretion: as VEGF secretion was increased time dependent manner in response to the hypoxia induction in TFF3-WT cells; however, VEGF production was significantly decreased in TFF3-KD cells (621 ± 89 vs. 264 ± 73 at 6 h and 969 ± 97 vs. 508 ± 69 at 12 h, P < 0.05). Our data demonstrated the TFF3 mediated regulation of VEGF expression induced by hypoxia, and implicated that TFF3 might be applied as a potential anti-angiogenic target for treatment of gastric cancer.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Abbreviations

VEGF:

Vascular endothelial growth factor

HIF:

Hypoxia-inducible transcription factors

IGF:

Insulin-like growth factor

HRE:

Hypoxia-response promoter elements

TFF:

Trefoil factor

KD:

Knockdown

ANOVA:

Analysis of variance

References

  1. Lazar D, Raica M, Sporea I, Taban S, Goldis A (2006) Cornianu M: tumor angiogenesis in gastric cancer. Rom J Morphol Embryol 47(1):5–13

    PubMed  Google Scholar 

  2. Willett CG, Boucher Y, di Tomaso E, Duda DG, Munn LL, Tong RT, Chung DC, Sahani DV, Kalva SP, Kozin SV et al (2004) Direct evidence that the VEGF-specific antibody bevacizumab has antivascular effects in human rectal cancer. Nat Med 10(2):145–147

    Article  PubMed  CAS  Google Scholar 

  3. Lyden D, Hattori K, Dias S, Costa C, Blaikie P, Butros L, Chadburn A, Heissig B, Marks W, Witte L et al (2001) Impaired recruitment of bone-marrow-derived endothelial and hematopoietic precursor cells blocks tumor angiogenesis and growth. Nat Med 7(11):1194–1201

    Article  PubMed  CAS  Google Scholar 

  4. Guleng B, Tateishi K, Ohta M, Kanai F, Jazag A, Ijichi H, Tanaka Y, Washida M, Morikane K, Fukushima Y (2005) Blockade of the stromal cell-derived factor-1/CXCR4 axis attenuates in vivo tumor growth by inhibiting angiogenesis in a vascular endothelial growth factor-independent manner. Cancer Res 65(13):5864–5871

    Article  PubMed  CAS  Google Scholar 

  5. Carmeliet P (2000) Collen D: molecular basis of angiogenesis Role of VEGF and VE-cadherin. Ann NY Acad Sci 902:249–262 Discussion 262–244

    Article  PubMed  CAS  Google Scholar 

  6. Carmeliet P (2003) Angiogenesis in health and disease. Nat Med 9(6):653–660

    Article  PubMed  CAS  Google Scholar 

  7. Carmeliet P, Dor Y, Herbert JM, Fukumura D, Brusselmans K, Dewerchin M, Neeman M, Bono F, Abramovitch R, Maxwell P et al (1998) Role of HIF-1alpha in hypoxia-mediated apoptosis, cell proliferation and tumour angiogenesis. Nature 394(6692):485–490

    Article  PubMed  CAS  Google Scholar 

  8. Yanyan C, Guoxian Q, Yang G, Leting W (2008) Mechanism of hypoxia-induced factor 1alpha expression in endothelial cells of the human umbilical vein and its induction of apoptosis. Mol Biol Rep 35(3):285–290

    Article  PubMed  Google Scholar 

  9. Jiang BH, Semenza GL, Bauer C, Marti HH (1996) Hypoxia-inducible factor 1 levels vary exponentially over a physiologically relevant range of O2 tension. Am J Physiol 271(4 Pt 1):C1172–C1180

    PubMed  CAS  Google Scholar 

  10. Lemus-Varela ML, Flores-Soto ME, Cervantes-Munguia R, Torres-Mendoza BM, Gudino-Cabrera G, Chaparro-Huerta V, Ortuno-Sahagun D, Beas-Zarate C (2010) Expression of HIF-1 alpha, VEGF and EPO in peripheral blood from patients with two cardiac abnormalities associated with hypoxia. Clin Biochem 43(3):234–239

    Article  PubMed  CAS  Google Scholar 

  11. Meyer zum Buschenfelde D, Hoschutzky H, Tauber R, Huber O (2004) Molecular mechanisms involved in TFF3 peptide-mediated modulation of the E-cadherin/catenin cell adhesion complex. Peptides 25(5):873–883

    Article  PubMed  CAS  Google Scholar 

  12. Podolsky DK (2000) Mechanisms of regulatory peptide action in the gastrointestinal tract: trefoil peptides. J Gastroenterol 35(Suppl 12):69–74

    PubMed  CAS  Google Scholar 

  13. Emami S, Rodrigues S, Rodrigue CM, Le Floch N, Rivat C, Attoub S, Bruyneel E, Gespach C (2004) Trefoil factor family (TFF) peptides and cancer progression. Peptides 25(5):885–898

    Article  PubMed  CAS  Google Scholar 

  14. Rivat C, Rodrigues S, Bruyneel E, Pietu G, Robert A, Redeuilh G, Bracke M, Gespach C, Attoub S (2005) Implication of STAT3 signaling in human colonic cancer cells during intestinal trefoil factor 3 (TFF3)—and vascular endothelial growth factor-mediated cellular invasion and tumor growth. Cancer Res 65(1):195–202

    PubMed  Google Scholar 

  15. Furuta GT, Turner JR, Taylor CT, Hershberg RM, Comerford K, Narravula S, Podolsky DK, Colgan SP (2001) Hypoxia-inducible factor 1-dependent induction of intestinal trefoil factor protects barrier function during hypoxia. J Exp Med 193(9):1027–1034

    Article  PubMed  CAS  Google Scholar 

  16. Louis NA, Hamilton KE, Canny G, Shekels LL, Ho SB, Colgan SP (2006) Selective induction of mucin-3 by hypoxia in intestinal epithelia. J Cell Biochem 99(6):1616–1627

    Article  PubMed  CAS  Google Scholar 

  17. Hernandez C, Santamatilde E, McCreath KJ, Cervera AM, Diez I, Ortiz-Masia D, Martinez N, Calatayud S, Esplugues JV, Barrachina MD (2009) Induction of trefoil factor (TFF)1, TFF2 and TFF3 by hypoxia is mediated by hypoxia inducible factor-1: implications for gastric mucosal healing. Br J Pharmacol 156(2):262–272

    Article  PubMed  CAS  Google Scholar 

  18. Liu FQ, Liu Y, Lui VC, Lamb JR, Tam PK, Chen Y (2008) Hypoxia modulates lipopolysaccharide induced TNF-alpha expression in murine macrophages. Exp Cell Res 314(6):1327–1336

    Article  PubMed  CAS  Google Scholar 

  19. Miyagishi M, Taira K (2002) U6 promoter-driven siRNAs with four uridine 3′ overhangs efficiently suppress targeted gene expression in mammalian cells. Nat Biotechnol 20(5):497–500

    Article  PubMed  CAS  Google Scholar 

  20. Jazag A, Ijichi H, Kanai F, Imamura T, Guleng B, Ohta M, Imamura J, Tanaka Y, Tateishi K, Ikenoue T et al (2005) Smad4 silencing in pancreatic cancer cell lines using stable RNA interference and gene expression profiles induced by transforming growth factor-beta. Oncogene 24(4):662–671

    Article  PubMed  CAS  Google Scholar 

  21. Guleng B, Tateishi K, Kanai F, Jazag A, Ohta M, Asaoka Y, Ijichi H, Tanaka Y, Imamura J, Ikenoue T et al (2005) Cancer-derived VEGF plays no role in malignant ascites formation in the mouse. World J Gastroenterol 11(35):5455–5459

    PubMed  CAS  Google Scholar 

  22. Shirato K, Kizaki T, Sakurai T, Ogasawara JE, Ishibashi Y, Iijima T, Okada C, Noguchi I, Imaizumi K, Taniguchi N et al (2009) Hypoxia-inducible factor-1alpha suppresses the expression of macrophage scavenger receptor 1. Pflugers Arch 459(1):93–103

    Article  PubMed  CAS  Google Scholar 

  23. Emami S, Le Floch N, Bruyneel E, Thim L, May F, Westley B, Rio M, Mareel M, Gespach C (2001) Induction of scattering and cellular invasion by trefoil peptides in src- and RhoA-transformed kidney and colonic epithelial cells. Faseb J 15(2):351–361

    Article  PubMed  CAS  Google Scholar 

  24. Baus-Loncar M, Giraud AS (2005) Multiple regulatory pathways for trefoil factor (TFF) genes. Cell Mol Life Sci 62(24):2921–2931

    Article  PubMed  CAS  Google Scholar 

  25. Loboda A, Jozkowicz A, Dulak J (2010) HIF-1 and HIF-2 transcription factors—similar but not identical. Mol Cells 29(5):435–442

    Article  PubMed  CAS  Google Scholar 

  26. Cummins EP, Taylor CT (2005) Hypoxia-responsive transcription factors. Pflugers Arch 450(6):363–371

    Article  PubMed  CAS  Google Scholar 

  27. De Maria R, Zeuner A, Eramo A, Domenichelli C, Bonci D, Grignani F, Srinivasula SM, Alnemri ES, Testa U, Peschle C (1999) Negative regulation of erythropoiesis by caspase-mediated cleavage of GATA-1. Nature 401(6752):489–493

    Article  PubMed  Google Scholar 

  28. Kramer MF, Gunaratne P, Ferreira GC (2000) Transcriptional regulation of the murine erythroid-specific 5-aminolevulinate synthase gene. Gene 247(1–2):153–166

    Article  PubMed  CAS  Google Scholar 

  29. Tarumoto T, Imagawa S, Ohmine K, Nagai T, Higuchi M, Imai N, Suzuki N, Yamamoto M, Ozawa K (2000) N(G)-monomethyl-l-arginine inhibits erythropoietin gene expression by stimulating GATA-2. Blood 96(5):1716–1722

    PubMed  CAS  Google Scholar 

  30. Chu SH, Feng DF, Ma YB, Zhu ZA, Zhang H, Qiu JH (2009) Stabilization of hepatocyte growth factor mRNA by hypoxia-inducible factor 1. Mol Biol Rep 36(7):1967–1975

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgment

This work was supported by National Natural Science Foundation of China (No. 30971362 & 81072013); Key Projects for Technology Plan of Fujian Province in China (No. 2009D020); Fundamental Research Funds for the Central Universities in China (No. 2010111082); and Foundations of Health Bureau of Fujian and Xiamen in China (No. 2007CXB8 & No. 3502z20077046).

Author information

Authors and Affiliations

  1. Department of Gastroenterology, Zhongshan Hospital Affiliated to Xiamen University, 201 Hubin South Road, Xiamen, 361004, Fujian, China

    Bayasi Guleng, Jia Han, Xiao-Ning Yang, Jing-Jing Liu & Jian-Lin Ren

  2. Medical College of Xiamen University, 168 University Road, Xiamen, 361005, Fujian, China

    Jin-Qiu Yang

  3. Central Laboratory, Zhongshan Hospital Affiliated to Xiamen University, 201 Hubin South Road, Xiamen, 361004, Fujian, China

    Qing-Wen Huang & Jian-Kun Huang

Authors
  1. Bayasi Guleng
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jia Han
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jin-Qiu Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Qing-Wen Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jian-Kun Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Xiao-Ning Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jing-Jing Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Jian-Lin Ren
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Bayasi Guleng or Jian-Lin Ren.

Additional information

Bayasi Guleng and Jia Han contributed equally to this work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

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 39, 4127–4134 (2012). https://doi.org/10.1007/s11033-011-1195-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11033-011-1195-2

Keywords

Search

Navigation