Cellular Oncology

, Volume 35, Issue 6, pp 461–471 | Cite as

High expression of heme oxygenase-1 is associated with tumor invasiveness and poor clinical outcome in non-small cell lung cancer patients

  • Jong-Rung Tsai
  • Hui-Min Wang
  • Po-Len Liu
  • Yung-Hsiang Chen
  • Ming-Chan Yang
  • Shah-Hwa Chou
  • Yu-Jen Cheng
  • Wei-Hsian Yin
  • Jhi-Jhu Hwang
  • Inn-Wen Chong
Original Papers

Abstract

Background

Heme oxygenase-1 (HO-1), a rate-limiting enzyme in heme catabolism, is known to play a role in the protection of cells against oxidative stress, inflammation, anomalous proliferation and apoptosis. As yet, the role of HO-1 expression in non-small cell lung cancer (NSCLC) development and metastasis remains unclear and insufficient data are available regarding its impact on the prognosis of NSCLC patients.

Methods

Seventy NSCLC patients who underwent surgical resection were included in this HO-1 expression study and, concomitantly, clinical parameters were collected. Two lung adenocarcinoma cell lines (A549 and H441) were used to assess both invasive and migratory parameters in vitro.

Results

NSCLC patients with a high HO-1 expression ratio (tumor tissue/normal tissue) (> 1) exhibited a significantly poorer prognosis and a higher metastatic rate compared to those with a low HO-1 expression ratio (p < 0.05). The invasive and migratory abilities of A549 and H441 cells significantly increased after exogenous HO-1 over-expression and significantly decreased after siRNA-mediated HO-1 expression silencing. HO-1 up- and down-regulation also positively correlated with the expression of metastasis-associated proteins EGFR, CD147 and MMP-9. In addition, we found that HO-1 expression can be inhibited by PI3K and AKT inhibitors, but not by MAPK inhibitors.

Conclusions

HO-1 is a poor prognostic NSCLC predictor and its over-expression may increase the metastatic potential of NSCLC. Based on our findings and those of others, HO-1 may be considered as a novel NSCLC therapeutic target.

Keywords

Heme Oxygenase-1 Metastasis Prognosis Non-small cell lung cancer 

Notes

Acknowledgements

This study was supported by grants from the National Science Council, grants NSC NSC99-2314-B-037-058-MY2 and NSC 100-2320-B-039-008-MY2, Kaohsiung Medical University Hospital grant KMUH99-9 M64, and Kaohsiung Medical University grant KMU-Q098022, Taiwan.

Disclosure/state of interest

The authors declare that they have no competing interests.

References

  1. 1.
    M.D. Maines, Heme oxygenase: Function, multiplicity, regulatory mechanisms, and clinical applications. FASEB J. 2, 2557–2568 (1988)PubMedGoogle Scholar
  2. 2.
    J.C. Becker, H. Fukui, Y. Imai, A. Sekikawa, T. Kimura, H. Yamagishi et al., Colonic expression of heme oxygenase-1 is associated with a better long-term survival in patients with colorectal cancer. Scand. J. Gastroenterol. 42, 852–858 (2007)PubMedCrossRefGoogle Scholar
  3. 3.
    M.D. Maines, The heme oxygenase system: A regulator of second messenger gases. Annu. Rev. Pharmacol. Toxicol. 37, 517–554 (1997)PubMedCrossRefGoogle Scholar
  4. 4.
    W. Durante, Heme oxygenase-1 in growth control and its clinical application to vascular disease. J. Cell. Physiol. 195, 373–382 (2003)PubMedCrossRefGoogle Scholar
  5. 5.
    D. Morse, A.M. Choi, Heme oxygenase-1: The “emerging molecule” has arrived. American journal of respiratory cell and molecular biology 27, 8–16 (2002)PubMedGoogle Scholar
  6. 6.
    S.S. Lee, S.F. Yang, C.H. Tsai, M.C. Chou, M.Y. Chou, Y.C. Chang, Upregulation of heme oxygenase-1 expression in areca-quid-chewing-associated oral squamous cell carcinoma. Journal of the Formosan Medical Association Taiwan yi zhi. 107, 355–363 (2008)PubMedCrossRefGoogle Scholar
  7. 7.
    S. Ishikawa, S. Tamaki, M. Ohata, K. Arihara, M. Itoh, Heme induces DNA damage and hyperproliferation of colonic epithelial cells via hydrogen peroxide produced by heme oxygenase: A possible mechanism of heme-induced colon cancer. Mol. Nutr. Food. Res. 54, 1182–1191 (2010)PubMedGoogle Scholar
  8. 8.
    T. Ishikawa, N. Yoshida, H. Higashihara, M. Inoue, K. Uchiyama, T. Takagi et al., Different effects of constitutive nitric oxide synthase and heme oxygenase on pulmonary or liver metastasis of colon cancer in mice. Clinical & experimental metastasis. 20, 445–450 (2003)CrossRefGoogle Scholar
  9. 9.
    G. Gueron, A. De Siervi, M. Ferrando, M. Salierno, P. De Luca, B. Elguero et al., Critical role of endogenous heme oxygenase 1 as a tuner of the invasive potential of prostate cancer cells. Mol. Cancer Res. 7, 1745–1755 (2009)PubMedCrossRefGoogle Scholar
  10. 10.
    M.A. Alaoui-Jamali, T.A. Bismar, A. Gupta, W.A. Szarek, J. Su, W. Song et al., A novel experimental heme oxygenase-1-targeted therapy for hormone-refractory prostate cancer. Cancer Res. 69, 8017–8024 (2009)PubMedCrossRefGoogle Scholar
  11. 11.
    H.R. Kim, S. Kim, E.J. Kim, J.H. Park, S.H. Yang, E.T. Jeong et al., Suppression of Nrf2-driven heme oxygenase-1 enhances the chemosensitivity of lung cancer A549 cells toward cisplatin. Lung Cancer 60, 47–56 (2008)PubMedCrossRefGoogle Scholar
  12. 12.
    K. Hirai, T. Sasahira, H. Ohmori, K. Fujii, H. Kuniyasu, Inhibition of heme oxygenase-1 by zinc protoporphyrin IX reduces tumor growth of LL/2 lung cancer in C57BL mice. Int. J. Cancer 120, 500–505 (2007)PubMedCrossRefGoogle Scholar
  13. 13.
    P. Boschetto, E. Zeni, L. Mazzetti, D. Miotto, N. Lo Cascio, P. Maestrelli et al., Decreased heme-oxygenase (HO)-1 in the macrophages of non-small cell lung cancer. Lung Cancer 59, 192–197 (2008)PubMedCrossRefGoogle Scholar
  14. 14.
    M.S. Degese, J.E. Mendizabal, N.A. Gandini, J.S. Gutkind, A. Molinolo, S.M. Hewitt et al., Expression of heme oxygenase-1 in non-small cell lung cancer (NSCLC) and its correlation with clinical data. Lung Cancer 77, 168–175 (2012)PubMedCrossRefGoogle Scholar
  15. 15.
    A. Jozkowicz, H. Was, J. Dulak, Heme oxygenase-1 in tumors: Is it a false friend? Antioxid. Redox. Signal. 9, 2099–2117 (2007)PubMedCrossRefGoogle Scholar
  16. 16.
    R.S. Herbst, Review of epidermal growth factor receptor biology. Int. J. Radiat. Oncol. Biol. Phys. 59, 21–26 (2004)PubMedCrossRefGoogle Scholar
  17. 17.
    H. Sasaki, H. Yukiue, K. Mizuno, A. Sekimura, A. Konishi, M. Yano et al., Elevated serum epidermal growth factor receptor level is correlated with lymph node metastasis in lung cancer. International journal of clinical oncology/Japan Society of Clinical Oncology. 8, 79–82 (2003)PubMedCrossRefGoogle Scholar
  18. 18.
    T. Kanekura, X. Chen, CD147/basigin promotes progression of malignant melanoma and other cancers. J. Dermatol. Sci. 57, 149–154 (2010)PubMedCrossRefGoogle Scholar
  19. 19.
    J. Xu, H.Y. Xu, Q. Zhang, F. Song, J.L. Jiang, X.M. Yang et al., HAb18G/CD147 functions in invasion and metastasis of hepatocellular carcinoma. Mol. Cancer Res. 5, 605–614 (2007)PubMedCrossRefGoogle Scholar
  20. 20.
    H. Voigt, C.S. Vetter-Kauczok, D. Schrama, U.B. Hofmann, J.C. Becker, R. Houben, CD147 impacts angiogenesis and metastasis formation. Cancer investigation. 27, 329–333 (2009)PubMedCrossRefGoogle Scholar
  21. 21.
    H. Was, J. Dulak, A. Jozkowicz, Heme oxygenase-1 in tumor biology and therapy. Current drug targets. 11, 1551–1570 (2010)PubMedCrossRefGoogle Scholar
  22. 22.
    H. Was, T. Cichon, R. Smolarczyk, D. Rudnicka, M. Stopa, C. Chevalier et al., Overexpression of heme oxygenase-1 in murine melanoma: Increased proliferation and viability of tumor cells, decreased survival of mice. Am. J. Pathol. 169, 2181–2198 (2006)PubMedCrossRefGoogle Scholar
  23. 23.
    M.H. Tsuji, T. Yanagawa, S. Iwasa, K. Tabuchi, K. Onizawa, S. Bannai et al., Heme oxygenase-1 expression in oral squamous cell carcinoma as involved in lymph node metastasis. Cancer Lett. 138, 53–59 (1999)PubMedCrossRefGoogle Scholar
  24. 24.
    M. Sunamura, D.G. Duda, M.H. Ghattas, L. Lozonschi, F. Motoi, J. Yamauchi et al., Heme oxygenase-1 accelerates tumor angiogenesis of human pancreatic cancer. Angiogenesis 6, 15–24 (2003)PubMedCrossRefGoogle Scholar
  25. 25.
    P. Boschetto, E. Zeni, L. Mazzetti, D. Miotto, N.L. Cascio, P. Maestrelli et al., Decreased heme-oxygenase (HO)-1 in the macrophages of non-small cell lung cancer. Lung Cancer 59, 192–197 (2008)PubMedCrossRefGoogle Scholar
  26. 26.
    C.K. Andreadi, L.M. Howells, P.A. Atherfold, M.M. Manson, Involvement of Nrf2, p38, B-Raf, and nuclear factor-kappaB, but not phosphatidylinositol 3-kinase, in induction of hemeoxygenase-1 by dietary polyphenols. Mol. Pharmacol. 69, 1033–1040 (2006)PubMedGoogle Scholar
  27. 27.
    S. Kocanova, E. Buytaert, J.Y. Matroule, J. Piette, J. Golab, P. de Witte et al., Induction of heme-oxygenase 1 requires the p38MAPK and PI3K pathways and suppresses apoptotic cell death following hypericin-mediated photodynamic therapy. Apoptosis 12, 731–741 (2007)PubMedCrossRefGoogle Scholar
  28. 28.
    E.J. Joung, M.H. Li, H.G. Lee, N. Somparn, Y.S. Jung, H.K. Na et al., Capsaicin induces heme oxygenase-1 expression in HepG2 cells via activation of PI3K-Nrf2 signaling: NAD(P)H:quinone oxidoreductase as a potential target. Antioxid. Redox. Signal. 9, 2087–2098 (2007)PubMedCrossRefGoogle Scholar
  29. 29.
    C.F. Mountain, Revisions in the International System for Staging Lung Cancer. Chest 111, 1710–1717 (1997)PubMedCrossRefGoogle Scholar
  30. 30.
    V.K. Sarhadi, H. Wikman, K. Salmenkivi, E. Kuosma, T. Sioris, J. Salo et al., Increased expression of high mobility group A proteins in lung cancer. J. Pathol. 209, 206–212 (2006)PubMedCrossRefGoogle Scholar
  31. 31.
    H. Wikman, E. Kettunen, J.K. Seppanen, A. Karjalainen, J. Hollmen, S. Anttila et al., Identification of differentially expressed genes in pulmonary adenocarcinoma by using cDNA array. Oncogene 21, 5804–5813 (2002)PubMedCrossRefGoogle Scholar
  32. 32.
    T.C. Wu, Y.H. Chen, H.B. Leu, Y.L. Chen, F.Y. Lin, S.J. Lin et al., Carvedilol, a pharmacological antioxidant, inhibits neointimal matrix metalloproteinase-2 and −9 in experimental atherosclerosis. Free Radic. Biol. Med. 43, 1508–1522 (2007)PubMedCrossRefGoogle Scholar
  33. 33.
    S.J. Lin, I.T. Lee, Y.H. Chen, F.Y. Lin, L.M. Sheu, H.H. Ku et al., Salvianolic acid B attenuates MMP-2 and MMP-9 expression in vivo in apolipoprotein-E-deficient mouse aorta and in vitro in LPS-treated human aortic smooth muscle cells. J. Cell. Biochem. 100, 372–384 (2007)PubMedCrossRefGoogle Scholar
  34. 34.
    H.M. Wang, C.C. Chiu, P.F. Wu, C.Y. Chen, Subamolide E from Cinnamomum subavenium induces sub-G1 cell-cycle arrest and caspase-dependent apoptosis and reduces the migration ability of human melanoma cells. J. Agric. Food Chem. 59, 8187–8192 (2011)PubMedCrossRefGoogle Scholar
  35. 35.
    F. Shiraishi, L.M. Curtis, L. Truong, K. Poss, G.A. Visner, K. Madsen, H.S. Nick, A. Agarwal, Heme oxygenase-1 gene ablation or expression modulates cisplatin-induced renal tubular apoptosis. Am J Physiol Renal Physiol 278, F726–F736 (2000)PubMedGoogle Scholar
  36. 36.
    A. Albini, R. Benelli, The chemoinvasion assay: A method to assess tumor and endothelial cell invasion and its modulation. Nat. Protoc. 2(504–11) (2007)Google Scholar
  37. 37.
    S. Tang, K.G. Morgan, C. Parker, J.A. Ware, Requirement for protein kinase C theta for cell cycle progression and formation of actin stress fibers and filopodia in vascular endothelial cells. J. Biol. Chem. 272, 28704–28711 (1997)PubMedCrossRefGoogle Scholar
  38. 38.
    G. Wang, E. Reed, Q.Q. Li, Molecular basis of cellular response to cisplatin chemotherapy in non-small cell lung cancer (Review). Oncol. Rep. 12, 955–965 (2004)PubMedGoogle Scholar
  39. 39.
    N. Grabinski, K. Bartkowiak, K. Grupp, B. Brandt, K. Pantel, M. Jucker, Distinct functional roles of Akt isoforms for proliferation, survival, migration and EGF-mediated signalling in lung cancer derived disseminated tumor cells. Cell. Signal. 23, 1952–1960 (2011)PubMedCrossRefGoogle Scholar
  40. 40.
    L. Mansi, E. Viel, E. Curtit, J. Medioni, C. Le Tourneau, Targeting the RAS signalling pathway in cancer therapy. Nat. Rev. Cancer 3, 11–22 (2003)CrossRefGoogle Scholar
  41. 41.
    R. Tenhunen, H.S. Marver, R. Schmid, The enzymatic conversion of heme to bilirubin by microsomal heme oxygenase. Proc. Natl. Acad. Sci. U. S. A. 61, 748–755 (1968)PubMedCrossRefGoogle Scholar
  42. 42.
    K. Doi, T. Akaike, S. Fujii, S. Tanaka, N. Ikebe, T. Beppu et al., Induction of haem oxygenase-1 nitric oxide and ischaemia in experimental solid tumours and implications for tumour growth. Br. J. Cancer 80, 1945–1954 (1999)PubMedCrossRefGoogle Scholar
  43. 43.
    E. Hara, K. Takahashi, T. Tominaga, T. Kumabe, T. Kayama, H. Suzuki et al., Expression of heme oxygenase and inducible nitric oxide synthase mRNA in human brain tumors. Biochem. Biophys. Res. Commun. 224, 153–158 (1996)PubMedCrossRefGoogle Scholar
  44. 44.
    M.Y. Li, J. Yip, M.K. Hsin, T.S. Mok, Y. Wu, M.J. Underwood et al., Haem oxygenase-1 plays a central role in NNK-mediated lung carcinogenesis. Eur. Respir. J. 32, 911–923 (2008)PubMedCrossRefGoogle Scholar
  45. 45.
    H. Kuroda, M. Takeno, S. Murakami, N. Miyazawa, T. Kaneko, Y. Ishigatsubo, Inhibition of heme oxygenase-1 with an epidermal growth factor receptor inhibitor and cisplatin decreases proliferation of lung cancer A549 cells. Lung Cancer 67, 31–36 (2010)PubMedCrossRefGoogle Scholar
  46. 46.
    P.L. Liu, J.R. Tsai, A.L. Charles, J.J. Hwang, S.H. Chou, Y.H. Ping et al., Resveratrol inhibits human lung adenocarcinoma cell metastasis by suppressing heme oxygenase 1-mediated nuclear factor-kappaB pathway and subsequently downregulating expression of matrix metalloproteinases. Mol. Nutr. Food Res. 54(2), S196–S204 (2010)PubMedCrossRefGoogle Scholar
  47. 47.
    T. Yanagawa, K. Omura, H. Harada, K. Nakaso, S. Iwasa, Y. Koyama et al., Heme oxygenase-1 expression predicts cervical lymph node metastasis of tongue squamous cell carcinomas. Oral oncology. 40, 21–27 (2004)PubMedCrossRefGoogle Scholar
  48. 48.
    G. De Palma, P. Mozzoni, O. Acampa, E. Internullo, P. Carbognani, M. Rusca et al., Expression levels of some antioxidant and epidermal growth factor receptor genes in patients with early-stage non-small cell lung cancer. Journal of nucleic acids (2010).Google Scholar
  49. 49.
    M. Mareel, I. Madani, Tumour-associated host cells participating at invasion and metastasis: Targets for therapy? Acta. chirurgica. Belgica. 106, 635–640 (2006)PubMedGoogle Scholar
  50. 50.
    A. Prawan, J.K. Kundu, Y.J. Surh, Molecular basis of heme oxygenase-1 induction: Implications for chemoprevention and chemoprotection. Antioxid. Redox. Signal. 7, 1688–1703 (2005)PubMedCrossRefGoogle Scholar
  51. 51.
    J. Alam, C. Wicks, D. Stewart, P. Gong, C. Touchard, S. Otterbein et al., Mechanism of heme oxygenase-1 gene activation by cadmium in MCF-7 mammary epithelial cells. Role of p38 kinase and Nrf2 transcription factor. J. Biol. Chem. 275, 27694–27702 (2000)PubMedGoogle Scholar
  52. 52.
    E.H. Kim, D.H. Kim, H.K. Na, Y.J. Surh, Effects of cyclopentenone prostaglandins on the expression of heme oxygenase-1 in MCF-7 cells. Ann. N. Y. Acad. Sci. 1030, 493–500 (2004)PubMedCrossRefGoogle Scholar

Copyright information

© International Society for Cellular Oncology 2012

Authors and Affiliations

  • Jong-Rung Tsai
    • 1
    • 2
  • Hui-Min Wang
    • 3
  • Po-Len Liu
    • 1
  • Yung-Hsiang Chen
    • 4
  • Ming-Chan Yang
    • 5
  • Shah-Hwa Chou
    • 1
    • 6
  • Yu-Jen Cheng
    • 7
  • Wei-Hsian Yin
    • 8
  • Jhi-Jhu Hwang
    • 1
    • 2
  • Inn-Wen Chong
    • 1
    • 2
  1. 1.Department of Respiratory TherapyCollege of Medicine, Kaohsiung Medical UniversityKaohsiungTaiwan
  2. 2.Department of Internal MedicineKaohsiung Medical University HospitalKaohsiungTaiwan
  3. 3.Department of Fragrance and Cosmetic ScienceKaohsiung Medical UniversityKaohsiungTaiwan
  4. 4.Graduate Institute of Integrated Medicine, College of Chinese Medicine, Department of UrologyChina Medical University and HospitalTaichungTaiwan
  5. 5.Department of SurgeryYuan’s General HospitalKaohsiungTaiwan
  6. 6.Department of Surgery, Division of Chest SurgeryKaohsiung Medical University HospitalKaohsiungTaiwan
  7. 7.Department of Health Management, Division of Thoracic SurgeryDepartment of Surgery, E-Da Hospital, I-Shou UniversityKaohsiungTaiwan
  8. 8.Division of CardiologyCheng-Hsin General Hospital, Faculty of Medicine, and Cardiovascular Research Centre, School of Medicine, National Yang-Ming UniversityTaipeiTaiwan

Personalised recommendations