SpringerLink
Log in

Cytochrome P450 ω-hydroxylase promotes angiogenesis and metastasis by upregulation of VEGF and MMP-9 in non-small cell lung cancer

  • Original Article
  • Published:
Cancer Chemotherapy and Pharmacology Aims and scope Submit manuscript

Abstract

Purpose

Cytochrome P450 (CYP) ω-hydroxylase, mainly consisting of CYP4A and CYP4F, converts arachidonic acid to 20-hydroxyeicosatetraenoic acid (20-HETE) that induces angiogenic responses in vivo and in vitro. The present study examined the role of CYP ω-hydroxylase in angiogenesis and metastasis of human non-small cell lung cancer (NSCLC).

Methods

The effect of WIT003, a stable 20-HETE analog, on invasion was evaluated using a modified Boyden chamber in three NSCLC cell lines. A549 cells were transfected with CYP4A11 expression vector or exposed to CYP ω-hydroxylase inhibitor (HET0016) or 20-HETE antagonist (WIT002), and then ω-hydroxylation activity toward arachidonic acid and the levels of matrix metalloproteinases (MMPs) and VEGF were detected. The in vivo effects of CYP ω-hydroxylase were tested in established tumor xenografts and an experimental metastasis model in athymic mice.

Results

Addition of WIT003 or overexpression of CYP4A11 with an associated increase in 20-HETE production significantly induced invasion and expression of VEGF and MMP-9. Treatment of A549 cells with HET0016 or WIT002 inhibited invasion with reduction in VEGF and MMP-9. The PI3 K or ERK inhibitors also attenuated expression of VEGF and MMP-9. Compared with control, CYP4A11 transfection significantly increased tumor weight, microvessel density (MVD), and lung metastasis by 2.5-fold, 2-fold, and 3-fold, respectively. In contrast, WIT002 or HET0016 decreased tumor volume, MVD, and spontaneous pulmonary metastasis occurrences.

Conclusion

CYP ω-hydroxylase promotes tumor angiogenesis and metastasis by upregulation of VEGF and MMP-9 via PI3 K and ERK1/2 signaling in human NSCLC cells.

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

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

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
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Granville CA, Dennis PA (2005) An overview of lung cancer genomics and proteomics. Am J Respir Cell Mol Biol 32:169

    Article  PubMed  CAS  Google Scholar 

  2. Miyata N, Roman RJ (2005) Role of 20-hydroxyeicosatetraenoic acid (20-HETE) in vascular system. J Smooth Muscle Res 41:175

    Article  PubMed  Google Scholar 

  3. Sun CW, Falck JR, Harder DR, Roman RJ (1999) Role of tyrosine kinase and PKC in the vasoconstrictor response to 20-HETE in renal arterioles. Hypertension 33:414

    PubMed  CAS  Google Scholar 

  4. Stec DE, Gannon KP, Beaird JS, Drummond HA (2007) 20-Hydroxyeicosatetraenoic acid (20-HETE) stimulates migration of vascular smooth muscle cells. Cell Physiol Biochem 19:121

    Article  PubMed  CAS  Google Scholar 

  5. Miyata N, Taniguchi K, Seki T, Ishimoto T, Sato-Watanabe M, Yasuda Y, Doi M, Kametani S, Tomishima Y, Ueki T, Sato MKameo K (2001) HET0016, a potent and selective inhibitor of 20-HETE synthesizing enzyme. Br J Pharmacol 133:325

    Article  PubMed  CAS  Google Scholar 

  6. Seki T, Wang MH, Miyata N, Laniado-Schwartzman M (2005) Cytochrome P450 4A isoform inhibitory profile of N-hydroxy-N’-(4-butyl-2-methylphenyl)-formamidine (HET0016), a selective inhibitor of 20-HETE synthesis. Biol Pharm Bull 28:1651

    Article  PubMed  CAS  Google Scholar 

  7. Chen P, Guo M, Wygle D, Edwards PA, Falck JR, Roman RJScicli AG (2005) Inhibitors of cytochrome P450 4A suppress angiogenic responses. Am J Pathol 166:615

    Article  PubMed  CAS  Google Scholar 

  8. Sandra LA, Kristopher GM, Daniela NS, Richard JR, Andrew SG (2003) CYP4A metabolites of arachidonic acid and VEGF are mediators of skeletal muscle angiogenesis. Am J Physiol Heart Circ Physiol 284:1528

    Google Scholar 

  9. Guo M, Roman RJ, Falck JR, Edwards PAScicli AG (2005) Human U251 glioma cell proliferation is suppressed by HET0016 [N-hydroxy-N’-(4-butyl-2-methylphenyl)formamidine], a selective inhibitor of CYP4A. J Pharmacol Exp Ther 315:526

    Article  PubMed  CAS  Google Scholar 

  10. Alexanian A, Rufanova VA, Miller B, Flasch A, Roman RJSorokin A (2009) Down-regulation of 20-HETE synthesis and signaling inhibits renal adenocarcinoma cell proliferation and tumor growth. Anticancer Res 29:3819

    PubMed  CAS  Google Scholar 

  11. Guo AM, Sheng J, Scicli GM, Arbab AS, Lehman NL, Edwards PA, Falck JR, Roman RJScicli AG (2008) Expression of CYP4A1 in U251 human glioma cell induces hyperproliferative phenotype in vitro and rapidly growing tumors in vivo. J Pharmacol Exp Ther 327:10

    Article  PubMed  CAS  Google Scholar 

  12. Guo AM, Arbab AS, Falck JR, Chen P, Edwards PA, Roman RJScicli AG (2007) Activation of vascular endothelial growth factor through reactive oxygen species mediates 20-hydroxyeicosatetraenoic acid-induced endothelial cell proliferation. J Pharmacol Exp Ther 321:18

    Article  PubMed  CAS  Google Scholar 

  13. Yamane M, Abe A (2000) Omega-hydroxylation activity toward leukotriene B(4) and polyunsaturated fatty acids in the human hepatoblastoma cell line, HepG2, and human lung adenocarcinoma cell line, A549. J Biochem 128:827

    PubMed  CAS  Google Scholar 

  14. Kadota K, Huang CL, Liu D, Ueno M, Kushida Y, Haba RYokomise H (2008) The clinical significance of lymphangiogenesis and angiogenesis in non-small cell lung cancer patients. Eur J Cancer 44:1057

    Article  PubMed  CAS  Google Scholar 

  15. Rasheed SA, Efferth T, Asangani IA, Allgayer H (2010) First evidence that the anti-malarial drug Artesunate inhibits invasion and in vivo metastasis in lung cancer by targeting essential extracellular proteases. Int J Cancer 127:1475

    Article  PubMed  CAS  Google Scholar 

  16. Shah SA, Spinale FG, Ikonomidis JS, Stroud RE, Chang EI, Reed CE (2010) Differential matrix metalloproteinase levels in adenocarcinoma and squamous cell carcinoma of the lung. J Thorac Cardiovasc Surg 139:984

    Article  PubMed  CAS  Google Scholar 

  17. Iniesta P, Moran A, De Juan C, Gomez A, Hernando F, Garcia-Aranda C, Frias C, Diaz-Lopez A, Rodriguez-Jimenez FJ, Balibrea JLBenito M (2007) Biological and clinical significance of MMP-2, MMP-9, TIMP-1 and TIMP-2 in non-small cell lung cancer. Oncol Rep 17:217

    PubMed  CAS  Google Scholar 

  18. Safranek J, Pesta M, Holubec L, Kulda V, Dreslerova J, Vrzalova J, Topolcan O, Pesek M, Finek J, Treska V (2009) Expression of MMP-7, MMP-9, TIMP-1 and TIMP-2 mRNA in lung tissue of patients with non-small cell lung cancer (NSCLC) and benign pulmonary disease. Anticancer Res 29:2513

    PubMed  CAS  Google Scholar 

  19. Hicklin DJ, Ellis LM (2005) Role of the vascular endothelial growth factor pathway in tumor growth and angiogenesis. J Clin Oncol 23:1011

    Article  PubMed  CAS  Google Scholar 

  20. Lee S, Jilani SM, Nikolova GV, Carpizo D, Iruela-Arispe ML (2005) Processing of VEGF-A by matrix metalloproteinases regulates bioavailability and vascular patterning in tumors. J Cell Biol 169:681

    Article  PubMed  CAS  Google Scholar 

  21. Houck KA, Leung DW, Rowland AM, Winer J, Ferrara N (1992) Dual regulation of vascular endothelial growth factor bioavailability by genetic and proteolytic mechanisms. J Biol Chem 267:26031

    PubMed  CAS  Google Scholar 

  22. Ortega N, L’Faqihi FE, Plouet J (1998) Control of vascular endothelial growth factor angiogenic activity by the extracellular matrix. Biol Cell 90:381

    Article  PubMed  CAS  Google Scholar 

  23. Deryugina EI, Quigley JP (2006) Matrix metalloproteinases and tumor metastasis. Cancer Metastasis Rev 25:9

    Article  PubMed  CAS  Google Scholar 

  24. Carmeliet P, Jain RK (2000) Angiogenesis in cancer and other diseases. Nature 407:249

    Article  PubMed  CAS  Google Scholar 

  25. Wang DS, Miller R, Shaw R, Shaw G (1996) The Pleckstrin homology domain of human βI∑II spectrin is targeted to the plasma membrane in vivo. Biochem Biophys Res Commun 225:420

    Article  PubMed  CAS  Google Scholar 

  26. Cai KX, Tse LY, Leung C, Tam PK, Xu R, Sham MH (2008) Suppression of lung tumor growth and metastasis in mice by adeno-associated virus-mediated expression of vasostatin. Clin Cancer Res 14:939

    Article  PubMed  CAS  Google Scholar 

  27. Doki Y, Murakami K, Yamaura T, Sugiyama S, Misaki T, Saiki I (1999) Mediastinal lymph node metastasis model by orthotopic intrapulmonary implantation of Lewis lung carcinoma cells in mice. Br J Cancer 79:1121

    Article  PubMed  CAS  Google Scholar 

  28. Lv X, Wan J, Yang J, Cheng H, Li Y, Ao Y, Peng R (2008) Cytochrome P450 omega- hydroxylase inhibition reduces cardiomyocyte apoptosis via activation of ERK1/2 signaling in rat myocardial ischemia-reperfusion. Eur J Pharmacol 596:118

    Article  PubMed  CAS  Google Scholar 

  29. Duffy MJ, Duggan C (2004) The urokinase plasminogen activator system: a rich source of tumour markers for the individualised management of patients with cancer. Clin Biochem 37:541

    Article  PubMed  CAS  Google Scholar 

  30. Rofstad EK, Mathiesen B, Kindem K, Galappathi K (2006) Acidic extracellular pH promotes experimental metastasis of human melanoma cells in athymic nude mice. Cancer Res 66:6699

    Article  PubMed  CAS  Google Scholar 

  31. Fidler IJ, Ellis LM (1994) The implications of angiogenesis for the biology and therapy of cancer metastasis. Cell 79:185

    Article  PubMed  CAS  Google Scholar 

  32. Lamoreaux WJ, Fitzgerald ME, Reiner A, Hasty KA, Charles ST (1998) Vascular endothelial growth factor increases release of gelatinase A and decreases release of tissue inhibitor of metalloproteinases by microvascular endothelial cells in vitro. Microvasc Res 55:29

    Article  PubMed  CAS  Google Scholar 

  33. Kaplan RN, Psaila B, Lyden D (2007) Niche-to-niche migration of bone-marrow-derived cells. Trends Mol Med 13:72

    Article  PubMed  CAS  Google Scholar 

  34. Guo M, Roman RJ, Fenstermacher JD, Brown SL, Falck JR, Arbab AS, Edwards PA, Scicli AG (2006) 9L gliosarcoma cell proliferation and tumor growth in rats are suppressed by N-hydroxy-N’-(4-butyl-2-methylphenol) formamidine (HET0016), a selective inhibitor of CYP4A. J Pharmacol Exp Ther 317:97

    Article  PubMed  CAS  Google Scholar 

  35. Hukkanen J, Lassila A, Päivärinta K, Valanne S, Sarpo S, Hakkola J, Pelkonen O, Raunio H (2000) Induction and regulation of xenobiotic-metabolizing cytochrome P450 s in the human A549 lung adenocarcinoma cell line. Am J Respir Cell Mol Biol 22:360

    PubMed  CAS  Google Scholar 

  36. Jiang BH, Liu LZ (2008) AKT signaling in regulating angiogenesis. Curr Cancer Drug Targets 8:19

    Article  PubMed  CAS  Google Scholar 

  37. Qiao M, Sheng S, Pardee AB (2008) Metastasis and AKT activation. Cell Cycle 7:2991

    Article  PubMed  CAS  Google Scholar 

  38. Reddy KB, Nabha SM, Atanaskova N (2003) Role of MAP kinase in tumor progression and invasion. Cancer Metastasis Rev 22:395

    Article  PubMed  CAS  Google Scholar 

  39. Dhanasekaran A, Bodiga S, Gruenloh S, Gao Y, Dunn L, Falck JR, Buonaccorsi JN, Medhora M, Jacobs ER (2009) 20-HETE increases survival and decreases apoptosis in pulmonary arteries and pulmonary artery endothelial cells. Am J Physiol Heart Circ Physiol 296:H777

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We greatly thank Dr. Jiancheng Tu and Dr. **e xianfei for their helpful discussions and reading of this manuscript. Grant support: National Nature Science Foundation Committee of China (No. 301162662), Science Foundation of Health Department of Hubei Province (No. JX3B14), National institutes of health Grant GM31278 (to J.R.F.), and the Robert A.Welch Foundation (to J.R.F.).

Author information

Authors and Affiliations

  1. Department of Pharmacology, School of Medicine, Wuhan University, Donghu Road, Wuhan, 430071, China

    Wei Yu, Li Chen, Ying Li & **g Yang

  2. School of Pharmacy, China Pharmaceutical University, Nan**g, 210009, China

    Yu-Qing Yang

  3. University of Texas Southwestern Medical Center, Dallas, TX, USA

    John R. Falck

  4. Department of Women’s Health Services, Gynecologic Oncology, Henry Ford Health System, Detroit, MI, USA

    Austin M. Guo

Authors
  1. Wei Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Li Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yu-Qing Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. John R. Falck
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Austin M. Guo
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ying Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. **g Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to **g Yang.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Yu, W., Chen, L., Yang, YQ. et al. Cytochrome P450 ω-hydroxylase promotes angiogenesis and metastasis by upregulation of VEGF and MMP-9 in non-small cell lung cancer. Cancer Chemother Pharmacol 68, 619–629 (2011). https://doi.org/10.1007/s00280-010-1521-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00280-010-1521-8

Keywords

Search

Navigation