Chemosensitization of Tumor Cells: Inactivation of Nuclear Factor-Kappa B Associated with Chemosensitivity in Melanoma Cells After Combination Treatment with E2F-1 and Doxorubicin

  • Hongying Hao
  • H. Sam Zhou
  • Kelly M. McMastersEmail author
Part of the Methods in Molecular Biology™ book series (MIMB, volume 542)


Combination chemotherapy has been shown to be more effective than single-agent therapy for many types of cancer, but both are known to induce drug resistance in cancer cells. Two major culprits in the development of this drug resistance are nuclear factor-κB (NF-κB) and the multidrug resistance (MDR) gene. For this reason, chemogene therapy is emerging as a viable alternative to conventional chemotherapy combinations. We have shown that transduction of the E2F-1 gene in melanoma cells markedly increases cell sensitivity to doxorubicin, thereby producing a synergistic effect on melanoma cell apoptosis. Our microarray results show that the NF-κB pathway and related genes undergo significant changes after the combined treatment of E2F-1 and doxorubicin. In fact, inactivation of NF-κB is associated with melanoma cell apoptosis induced by E2F-1 and doxorubicin, providing a link between the NF-κB signaling pathway and the chemosensitivity of melanoma cells after this treatment.

Key Words

Apoptosis doxorubicin E2F-1 electrophoretic mobility shift assay (EMSA) nuclear factor-κB (NF-κB) 



We are grateful to Mrs. Margaret Abby for her expert manuscript editing. Supported by NIH Grant R01CA90784 to KMM.


  1. 1.
    Verweij, J. and de Jonge, M.J. (2000) Achievements and future of chemotherapy. Eur. J. Cancer 36, 1479–1487.PubMedCrossRefGoogle Scholar
  2. 2.
    Liem, A.A., Chamberlain, M.P., Wolf. C.R., and Thompson, A.M. (2002) The role of signal transduction in cancer treatment and drug resistance. Eur. J. Surg. Oncol. 28, 679–684.PubMedCrossRefGoogle Scholar
  3. 3.
    Meng, R.D., Phillips, P., and El-Deiry, W.S. (1999) p53-independent increase in E2F-1 expression enhances the cytotoxic effects of etoposide and of adriamycin. Int. J. Oncol. 14, 5–14.PubMedGoogle Scholar
  4. 4.
    Cheon, J., Ko, S.C., Gardner, T.A., Shirakawa, T., Gotoh, A., Kao, C., and Chung L.W. (1997) Chemogene therapy: osteocalcin promoter-based suicide gene therapy in combination with methotrexate in a murine osteosarcoma model. Cancer Gene. Ther. 4, 359–365.PubMedGoogle Scholar
  5. 5.
    Nemunaitis, J., Swisher, S.G., Timmons, T., Connors, D., Mack, M., Doerksen, L., Weill, D., Wait, J., Lawrence, D.D., Kemp, B.L., Fossella, F., Glisson, B.S., Hong, W.K., Khuri, F.R., Kurie, J.M., Lee, J.J. Lee, J.S., Nguyen, D.M., Nesbitt, J.C., Perez-Soler, R., Pisters, K.M.W., Putnam, J.B., Richli, W.R., Shin, D.M., Walsh, G.L., Merritt, J., and Roth, J. (2000) Adenovirus-mediated p53 gene transfer in sequence with cisplatin to tumors of patients with non-small-cell lung cancer. J. Clin. Oncol. 18, 609–622.PubMedGoogle Scholar
  6. 6.
    Yeh, P.Y., Chuang, S.E., Yeh, K.H., Song, Y.C., Ea, C.K., and Cheng, A.L. (2002) Increase of the resistance of human cervical carcinoma cells to cisplatin by inhibition of the MEK to ERK signaling pathway partly via enhancement of anticancer drug-induced NF-κB activation. Biochem. Pharmacol. 63, 1423–1430.PubMedCrossRefGoogle Scholar
  7. 7.
    Yeh, P.Y., Chuang, S.E., Yeh, K.H., Song, Y.C., and Cheng, A.L. (2003) Involvement of nuclear transcription factor-κB in lowdose doxorubicin-induced drug resistance of cervical carcinoma cells. Biochem. Pharmacol. 66, 25–33.PubMedCrossRefGoogle Scholar
  8. 8.
    Karin, M. (2006) Nuclear factor-kappaB in cancer development and progression. Nature 441(7092), 431–436.PubMedCrossRefGoogle Scholar
  9. 9.
    Ghosh, S. and Karin, M. (2002) Missing pieces in the NF-κB puzzle. Cell, 109(Suppl.), S81–S96.PubMedCrossRefGoogle Scholar
  10. 10.
    Rothwarf, D.M. and Karin, M. (1999) The NF-κB activation pathway: a paradigm in information transfer from membrane to nucleus. Sci. STKE (October 26) 1999(5), RE1.Google Scholar
  11. 11.
    Karin, M. and Ben-Neriah, Y. (2000) Phosphorylation meets ubiquitination: the control of NF-κB activity. Annu. Rev. Immunol. 18, 621–663.PubMedCrossRefGoogle Scholar
  12. 12.
    Werner, S.L., Barken, D. and Hoffmann, A. (2005) Stimulus specificity of gene expression programs determined by temporal control of IKK activity. Science. 309, 1857–1861.PubMedCrossRefGoogle Scholar
  13. 13.
    Park, J.M., Greten, F.R., Wong, A., Westrick, R.J., Arthur, S.C., Otsu, K., Hoffmann, A., Montminy, M. and Karin, M. (2005) Signaling pathways and genes that inhibit pathogen-induced macrophage apoptosis: CREB and NF-κB as key regulators. Immunity 23, 319–329.PubMedCrossRefGoogle Scholar
  14. 14.
    Covert, M.W., Leung, T.H., Gaston, J.E. and Baltimore, D. (2005) Achieving stability of lipopolysaccharide-induced NF-κB activation. Science 309, 1854–1857.PubMedCrossRefGoogle Scholar
  15. 15.
    Aggarwal, B.B. (2004) Nuclear factor-κB: the enemy within. Cancer Cell 6, 203–208.PubMedCrossRefGoogle Scholar
  16. 16.
    Hao, H., Dong, Y.B., Bowling, M.T., Zhou, H.S. and McMasters, K.M. (2006) Alteration of gene expression in melanoma cells following combined treatment with E2F-1 and adriamycin. Anticancer Res. 26(3A), 1947–1956.PubMedGoogle Scholar

Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  1. 1.Division of Surgical Oncology, Department of SurgeryUniversity of Louisville School of MedicineLouisvilleUSA

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