Annals of Surgical Oncology

, Volume 16, Issue 5, pp 1403–1411 | Cite as

Combined IFN-γ–Endostatin Gene Therapy and Radiotherapy Attenuates Primary Breast Tumor Growth and Lung Metastases via Enhanced CTL and NK Cell Activation and Attenuated Tumor Angiogenesis in a Murine Model

  • Lin Lin Liu
  • Myles J. Smith
  • Bao Sheng Sun
  • Guan Jun Wang
  • H. Paul Redmond
  • Jiang Huai Wang
Breast Oncology

Abstract

Background

Gene–radiotherapy, a combination of gene therapy and radiotherapy, is a new paradigm for cancer treatment, with the potential to simultaneously improve local and systemic breast cancer control. The aim of this study was to evaluate antitumor effect of interferon (IFN)-γ-endostatin-based gene–radiotherapy in a murine metastatic breast tumor model, and to elucidate possible mechanisms involved.

Methods

Murine mammary adenocarcinoma 4T1 cells transfected with pEgr-IFN-γ and pEgr-endostatin plasmids were irradiated (2–20 Gy). IFN-γ and endostatin levels in the culture supernatants were measured. In vivo female BALB/c mice were inoculated with 1 × 105 4T1 cells by mammary fat pad injection and divided into control, empty vector, gene therapy (pEgr-IFN-γ and pEgr-endostatin), radiotherapy, and combined gene–radiotherapy groups. Tumor growth, tumor/body weight ratio, lung metastases, and survival of the tumor-bearing mice were observed. Splenic cytotoxic T-lymphocyte (CTL) and natural killer (NK) cell activity and intratumor microvessel density were also assessed.

Results

Irradiation significantly enhanced IFN-γ and endostatin secretion from the transfected 4T1 cells. In vivo mice that received combined gene–radiotherapy showed maximal attenuation in tumor growth rate and lung metastases with increased survival compared with mice that received gene therapy or radiotherapy alone. This was associated with significantly enhanced CTL and NK cell activity and reduced intratumor microvessel density.

Conclusion

These results demonstrate that IFN-γ-endostatin-based gene–radiotherapy provide a potent antitumor effect in a murine metastatic breast tumor model, which may relate to IFN-γ-stimulated CTL and NK cell activation, and endostatin-induced antiangiogenic activity. Thus, gene–radiotherapy may represent a useful addition to neoadjuvant management of locally advanced breast cancer.

References

  1. 1.
    Buchholz TA, Hill BS, Tucker SL, Frye DK, Kuerer HM, Buzdar AU, et al. Factors predictive of outcome in patients with breast cancer refractory to neoadjuvant chemotherapy. Cancer J. 2001;7:413–20.PubMedGoogle Scholar
  2. 2.
    Yang J, Jin G, Liu X, Liu S. Therapeutic effect of pEgr-IL18-B7.2 gene radiotherapy in B16 melanoma-bearing mice. Hum Gene Ther. 2007;18:323–32.PubMedCrossRefGoogle Scholar
  3. 3.
    Jin GH, Jin SZ, Liu Y, Xu RM, Yang JZ, Pan XN, et al. Therapeutic effect of gene-therapy in combination with local X-irradiation in a mouse malignant melanoma model. Biochem Biophys Res Commun. 2005;330:975–81.PubMedCrossRefGoogle Scholar
  4. 4.
    Cioca DP, Deak E, Cioca F, Paunescu V. Monoclonal antibodies targeted against melanoma and ovarian tumors enhance dendritic cell-mediated cross-presentation of tumor-associated antigens and efficiently cross-prime CD8+ T cells. J Immunother. 2006;29:41–52.PubMedCrossRefGoogle Scholar
  5. 5.
    Toh U, Fujii T, Seki N, Niiya F, Shirouzu K, Yamana H. Characterization of IL-2-activated TILs and their use in intrapericardial immunotherapy in malignant pericardial effusion. Cancer Immunol Immunother. 2006;55;1219–27.PubMedCrossRefGoogle Scholar
  6. 6.
    Shilling DA, Smith MJ, Tyther R, Sheehan D, England K, Kavanagh EG, et al. Salmonella typhimurium stimulation combined with tumour-derived heat shock proteins induces potent dendritic cell anti-tumour responses in a murine model. Clin Exp Immunol. 2007;149:109–16.PubMedGoogle Scholar
  7. 7.
    Smith MJ, Culhane AC, Killeen S, Kelly MA, Wang JH, Cotter TG, et al. Mechanisms driving local breast cancer recurrence in a model of breast-conserving surgery. Ann Surg Oncol. 2008;15:2954–64.PubMedCrossRefGoogle Scholar
  8. 8.
    Hunder NN, Wallen H, Cao J, Hendricks DW, Reilly JZ, Rodmyre R, et al. Treatment of metastatic melanoma with autologous CD4+ T cells against NY-ESO-1. N Engl J Med. 2008;358:2698–703.PubMedCrossRefGoogle Scholar
  9. 9.
    Boehm U, Klamp T, Groot M, Howard JC. Cellular responses to interferon-gamma. Annu Rev Immunol. 1997;15:749–95 (Review).PubMedCrossRefGoogle Scholar
  10. 10.
    Lee SH, Aggarwal BB, Rinderknecht E, Assisi F, Chiu H. The synergistic anti-proliferative effect of gamma-interferon and human lymphotoxin. J Immunol. 1984;133:1083–86.PubMedGoogle Scholar
  11. 11.
    Bernstein W, Zou ZQ, Black RJ, Pirollo KF, Chang EH. Association of interferon-gamma induced growth inhibition and modulation of epidermal growth factor receptor gene expression in squamous cell carcinoma cell lines. J Biol Regul Homeost Agents. 1988;2:186–92.PubMedGoogle Scholar
  12. 12.
    Shi W, Siemann DW. Inhibition of renal cell carcinoma angiogenesis and growth by antisense oligonucleotides targeting vascular endothelia growth factor. Br J Cancer. 2002;87:119–26.PubMedCrossRefGoogle Scholar
  13. 13.
    Solorzano CC, Baker CH, Bruns CJ, Killion JJ, Ellis LM, Wood J, et al. Inhibition of growth and metastasis of human pancreatic cancer growing in nude mice by PTK787/ZK222584, an inhibitor of the vascular endothelia growth factor receptor tyrosine kinases. Cancer Biother Radiopharm. 2001;16:359–70.PubMedCrossRefGoogle Scholar
  14. 14.
    Zheng AQ, Song XR, Yu JM, Wei L, Wang XW. Liposome transfected to plasmid-encoding endostatin gene combined with radiotherapy inhibits liver cancer growth in nude mice. World J Gastroenterol. 2005;11:4439–42.PubMedGoogle Scholar
  15. 15.
    Shi W, Teschendorf C, Muzyczka N, Siemann DW. Adeno-associated virus-mediated gene transfer of endostatin inhibits angiogenesis and tumor growth in vivo. Cancer Gene Ther. 2002;9:513–21.PubMedCrossRefGoogle Scholar
  16. 16.
    Liekens S, De Clercq E, Neyts J. Angiogenesis: regulators and clinical applications. Biochem Pharmacol. 2001;61:253–70 (Review).PubMedCrossRefGoogle Scholar
  17. 17.
    Blezinger P, Wang J, Gondo M, Quezada A, Mehrens D, French M, et al. Systemic inhibition of tumor growth and tumor metastases by intramuscular administration of the endostatin gene. Nat Biotechnol. 1999;17:343–8.PubMedCrossRefGoogle Scholar
  18. 18.
    O’Reilly MS, Boehm T, Shing Y, Fukai N, Vasios G, Lane WS, et al. Endostatin: an endogenous inhibitor of angiogenesis and tumor growth. Cell. 1997;88:277–85.PubMedCrossRefGoogle Scholar
  19. 19.
    Folkman J. Angiogenesis in cancer, vascular, rheumatoid and other disease. Nat Med. 1995;1:27–31 (Review).PubMedCrossRefGoogle Scholar
  20. 20.
    Folkman J. Antiangiogenic gene therapy. Proc Natl Acad Sci USA. 1998;95:9064–6.PubMedCrossRefGoogle Scholar
  21. 21.
    Burke PA, DeNardo SJ. Antiangiogenic agents and their promising potential in combined therapy. Crit Rev Oncol Hematol. 2001;39:155–71 (Review).PubMedCrossRefGoogle Scholar
  22. 22.
    Sukhatme VP. Early transcriptional events in cell growth: the Egr family. J Am Soc Nephrol. 1990;1:859–66 (Review).PubMedGoogle Scholar
  23. 23.
    Cao XM, Koski RA, Gashler A, McKiernan M, Morris CF, Gaffney R, et al. Identification and characterization of the Egr-1 gene product, a DNA-binding zinc finger protein induced by differentiation and growth signals. Mol Cell Biol. 1990;10:1931–9.PubMedGoogle Scholar
  24. 24.
    Yang W, Li X. Anti-tumor effect of pEgr-interferon-gamma-endostatin gene-radiotherapy in mice bearing Lewis lung carcinoma and its mechanism. Chin Med J. 2005;118:296–301.PubMedGoogle Scholar
  25. 25.
    Weidner N, Gasparini G. Determination of epidermal growth factor receptor provides additional prognostic information to measuring tumor angiogenesis in breast carcinoma patients. Breast Cancer Res Treat. 1994;29: 97–107.PubMedCrossRefGoogle Scholar
  26. 26.
    Kwong DL, Sham JS, Leung LH, Cheng AC, Ng WM, Kwong PW, et al. Preliminary results of radiation dose escalation for locally advanced nasopharyngeal carcinoma. Int J Radiat Oncol Biol Phys. 2006;64:374–81.PubMedGoogle Scholar
  27. 27.
    Harari PM, Huang SM. Head and neck cancer as a clinical model for molecular targeting of therapy: combining EGFR blockade with radiation. Int J Radiat Oncol Biol Phys. 2001;49:427–33.PubMedCrossRefGoogle Scholar
  28. 28.
    Garnett KE, Chapman P, Chambers JA, Waddell ID, Boam DS. Differential gene expression between Zucker Fatty rats and Zucker Diabetic Fatty rats: a potential role for the immediate-early gene Egr–1 in regulation of beta cell proliferation. J Mol Endocrinol. 2005;35:13–25.PubMedCrossRefGoogle Scholar
  29. 29.
    Min FL, Zhang H, Li WJ. Current status of tumor radiogenic therapy. World J Gastroenterol. 2005;11:3014–9 (Review).PubMedGoogle Scholar
  30. 30.
    Lopez CA, Kimchi ET, Mauceri HJ, Park JO, Mehta N, Murphy KT, et al. Chemoinducible gene therapy: a strategy to enhance doxorubicin antitumor activity. Mol Cancer Ther. 2004;3:1167–75.PubMedCrossRefGoogle Scholar
  31. 31.
    Hahnfeldt P, Panigrahy D, Folkman J, Hlatky L. Tumor development under angiogenic signaling: a dynamical theory of tumor growth, treatment response, and postvascular dormancy. Cancer Res. 1999;59:4770–5.PubMedGoogle Scholar
  32. 32.
    Holmgren L, O’Reilly MS, Folkman J. Dormancy of micrometastases: balance proliferation and apoptosis in the presence of angiogenesis suppression. Nat Med. 1995;1:149–53.PubMedCrossRefGoogle Scholar
  33. 33.
    Ding I, Sun JZ, Fenton B, Liu WM, Kimsely P, Okunieff P, et al. Intratumoral administration of endostatin plasmid inhibits vascular growth and perfusion in MCa-4 murine mammary carcinomas. Cancer Res. 2001;61:526–31.PubMedGoogle Scholar
  34. 34.
    Nakashima Y, Yano M, Kobayashi Y, Moriyama S, Sasaki H, Toyama T, et al. Endostatin gene therapy on murine lung metastases model utilizing cationic vector-mediated intravenous gene delivery. Gene Ther. 2003;10:123–30.PubMedCrossRefGoogle Scholar
  35. 35.
    Chen QR, Kumar D, Stass SA, Mixson AJ. Liposomes complexed to plasmids encoding angiostatin and endostatin inhibit breast cancer in nude mice. Cancer Res. 1999;59:3308–12.PubMedGoogle Scholar
  36. 36.
    Nayak SK, McCallister T, Han LJ, Gangavalli R, Barber J, Dillman RO. Transduction of human renal carcinoma cells with human gamma-interferon gene via retroviral vector. Cancer Gene Ther. 1996;3:143–50.PubMedGoogle Scholar

Copyright information

© Society of Surgical Oncology 2009

Authors and Affiliations

  • Lin Lin Liu
    • 1
    • 2
  • Myles J. Smith
    • 1
  • Bao Sheng Sun
    • 1
    • 2
  • Guan Jun Wang
    • 2
  • H. Paul Redmond
    • 1
  • Jiang Huai Wang
    • 1
  1. 1.Department of Academic SurgeryUniversity College Cork (UCC)/National University of Ireland (NUI), Cork University HospitalCorkIreland
  2. 2.Department of Radiation Oncology, The Second Affiliated HospitalJilin UniversityChangchunPeople’s Republic of China

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