Apoptosis

, Volume 22, Issue 12, pp 1553–1563 | Cite as

Bromoethylindole (BEI-9) redirects NF-κB signaling induced by camptothecin and TNFα to promote cell death in colon cancer cells

  • Rupak Chowdhury
  • Dominique Gales
  • Paloma Valenzuela
  • Sonni Miller
  • Teshome Yehualaeshet
  • Upender Manne
  • Giulio Francia
  • Temesgen Samuel
Original Paper
  • 57 Downloads

Abstract

Chemotherapeutic regimens containing camptothecin (CPT), 5-fluorouracil, and oxaliplatin are used to treat advanced colorectal cancer. We previously reported that an indole derivative, 3-(2-bromoethyl)indole (BEI-9), inhibited the proliferation of colon cancer cells and suppressed NF-κB activation. Here, we show that a combination of BEI-9 with either CPT or tumor necrosis factor alpha (TNFα) enhances cell death. Using colorectal cancer cells, we examined the activation of NF-κB by drugs, the potential of BEI-9 for inhibiting drug-induced NF-κB activation, and the enhancement of cell death by combination treatments. Cells were treated with the chemotherapeutic drugs alone or in combination with BEI-9. NF-κB activation, cell cycle profiles, DNA-damage response, markers of cell death signaling and targets of NF-κB were evaluated to determine the effects of single and co-treatments. The combination of BEI-9 with CPT or TNFα inhibited NF-κB activation and reduced the expression of NF-κB-responsive genes, Bcl-xL and COX2. Compared to CPT or BEI-9 alone, sequential treatment of the cells with CPT and BEI-9 significantly enhanced caspase activation and cell death. Co-treatment with TNFα and BEI-9 also caused more cytotoxicity than TNFα or BEI-9 alone. Combined BEI-9 and TNFα enhanced cell death through caspase activation and cleavage of the switch-protein, RIP1 kinase. BEI-9 reduced the expression of COX2 both alone and in combination with CPT or TNF. We postulate that BEI-9 enhances the effects of these drugs on cancer cells by turning off or redirecting NF-κB signaling. Therefore, the combination of BEI-9 with drugs that activate NF-κB needs to be evaluated for clinical applications.

Keywords

Bromoethylindole Apoptosis NF-kB Camptothecin Colorectal cancer 

Supplementary material

10495_2017_1427_MOESM1_ESM.pptx (17.6 mb)
Supplementary Fig. 1: A. Photomicrographs of SW620 cells treated with BEI (1, 2, or 5 µM), TNFα, or with BEI plus TNF combinations as shown. Arrows point to cells showing membrane blebbing phenotypes suggestive of apoptosis. B. Phase contrast (upper row) and fluorescent images (lower row) of SW620 cells treated as shown. The spots in the lower row show pseudo-colored images of caspase-positive (FITC-VAD-FMK reactive) cells, which fluoresce green. In this experiment, CPT+BEI treatment was simultaneous, and did not result in enhanced cell death. Supplementary Fig. 2: A. Morphology of LoVo cells treated for 24 hr with vehicle, BEI, TNF, or a combination of the two drugs. B. A caspase 3/7 activation fluorescent assay that shows the caspase-activating effect of the TNFα – BEI combination. C. Morphology of Colo205 cells treated as shown. D. Cell cycle analysis of Colo205 cells treated as in panel C. The bars indicate the percentages of cells at each phase of the cell cycle shown on the X-axis. The percentage of sub-G1 cells increases with BEI and with the combination BEI and TNFα. Supplemental Fig. 3: Annexin -V/PI staining profiles of HCT116 cells treated as shown for 24 hr. The percentages of Annexin V-positive or Annexin -V/PI double-positive cells are increased by combination treatment compared to single agent treatments. (PPTX 18005 KB)

References

  1. 1.
    Deng Y, Cai Y, Lin J, Jiang L, Hu H (2015) Survival of patients with KRAS wild-type metastatic colorectal cancer is identical after sequential treatment with cetuximab and bevacizumab regardless of the sequence—a retrospective single-center study. Gastroenterol Rep 3:339–343Google Scholar
  2. 2.
    Kirstein MM, Lange A, Prenzler A, Manns MP, Kubicka S, Vogel A (2014) Targeted therapies in metastatic colorectal cancer: a systematic review and assessment of currently available data. Oncologist 19:1156–1168CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Samuel T, Fadlalla K, Gales DN, Putcha BD, Manne U (2014) Variable NF-kappaB pathway responses in colon cancer cells treated with chemotherapeutic drugs. BMC Cancer 14:599CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Huang TT, Wuerzberger-Davis SM, Seufzer BJ et al (2000) NF-kappaB activation by camptothecin. A linkage between nuclear DNA damage and cytoplasmic signaling events. J Biol Chem 275:9501–9509CrossRefPubMedGoogle Scholar
  5. 5.
    McCool KW, Miyamoto S (2012) DNA damage-dependent NF-kappaB activation: NEMO turns nuclear signaling inside out. Immunol Rev 246:311–326CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Brach MA, Hass R, Sherman ML, Gunji H, Weichselbaum R, Kufe D (1991) Ionizing radiation induces expression and binding activity of the nuclear factor kappa B. J Clin Invest 88:691–695CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Criswell T, Leskov K, Miyamoto S, Luo G, Boothman DA (2003) Transcription factors activated in mammalian cells after clinically relevant doses of ionizing radiation. Oncogene 22:5813–5827CrossRefPubMedGoogle Scholar
  8. 8.
    Prasad AV, Mohan N, Chandrasekar B, Meltz ML (1994) Activation of nuclear factor kappa B in human lymphoblastoid cells by low-dose ionizing radiation. Radiat Res 138:367–372CrossRefPubMedGoogle Scholar
  9. 9.
    Fadlalla K, Elgendy R, Gilbreath E et al (2015) 3-(2-Bromoethyl)-indole inhibits the growth of cancer cells and NF-kappaB activation. Oncol Rep 34:495–503CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Varfolomeev E, Vucic D. (2016) Intracellular regulation of TNF activity in health and disease. CytokineGoogle Scholar
  11. 11.
    Sun J, Han J, Zhu Q, Li Z, Hu J (2012) Camptothecin fails to induce apoptosis in tumor necrosis factor-alpha-treated HaCaT cells. Pharmacology 89:58–63CrossRefPubMedGoogle Scholar
  12. 12.
    Wu ZH, Shi Y, Tibbetts RS, Miyamoto S (2006) Molecular linkage between the kinase ATM and NF-kappaB signaling in response to genotoxic stimuli. Science 311:1141–1146CrossRefPubMedGoogle Scholar
  13. 13.
    Grivennikov SI, Kuprash DV, Liu ZG, Nedospasov SA (2006) Intracellular signals and events activated by cytokines of the tumor necrosis factor superfamily: from simple paradigms to complex mechanisms. Int Rev Cytol 252:129–161CrossRefPubMedGoogle Scholar
  14. 14.
    Baud V, Karin M (2001) Signal transduction by tumor necrosis factor and its relatives. Trends Cell Biol 11:372–377CrossRefPubMedGoogle Scholar
  15. 15.
    Aggarwal BB (2000) Tumour necrosis factors receptor associated signalling molecules and their role in activation of apoptosis, JNK and NF-kappaB. Ann Rheum Dis 59(Suppl 1):i6-16PubMedGoogle Scholar
  16. 16.
    Ray Chaudhuri A, Hashimoto Y, Herrador R et al (2012) Topoisomerase I poisoning results in PARP-mediated replication fork reversal. Nat Struct Mol Biol 19:417–423CrossRefPubMedGoogle Scholar
  17. 17.
    Dabkeviciene D, Jonusiene V, Zitkute V et al (2015) The role of interleukin-8 (CXCL8) and CXCR2 in acquired chemoresistance of human colorectal carcinoma cells HCT116. Medical Oncol 32:258CrossRefGoogle Scholar
  18. 18.
    Jamieson T, Clarke M, Steele CW et al (2012) Inhibition of CXCR2 profoundly suppresses inflammation-driven and spontaneous tumorigenesis. J Clin Invest 122:3127–3144CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Lee YS, Choi I, Ning Y et al (2012) Interleukin-8 and its receptor CXCR2 in the tumour microenvironment promote colon cancer growth, progression and metastasis. Br J Cancer 106:1833–1841CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Li L, Xu L, Yan J et al (2015) CXCR2-CXCL1 axis is correlated with neutrophil infiltration and predicts a poor prognosis in hepatocellular carcinoma. J Exp Clin Cancer Res 34:129CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Saintigny P, Massarelli E, Lin S et al (2013) CXCR2 expression in tumor cells is a poor prognostic factor and promotes invasion and metastasis in lung adenocarcinoma. Cancer Res 73:571–582CrossRefPubMedGoogle Scholar
  22. 22.
    Magnusson C, Vaux DL (1999) Signalling by CD95 and TNF receptors: not only life and death. Immunol Cell Biol 77:41–46CrossRefPubMedGoogle Scholar
  23. 23.
    Chu WM (2013) Tumor necrosis factor. Cancer Lett 328:222–225CrossRefPubMedGoogle Scholar
  24. 24.
    Lin Y, Devin A, Rodriguez Y, Liu ZG (1999) Cleavage of the death domain kinase RIP by caspase-8 prompts TNF-induced apoptosis. Genes Dev 13:2514–2526CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    He L, Kim BY, Kim KA et al (2007) NF-kappaB inhibition enhances caspase-3 degradation of Akt1 and apoptosis in response to camptothecin. Cell Signal 19:1713–1721CrossRefPubMedGoogle Scholar
  26. 26.
    Togano T, Sasaki M, Watanabe M et al (2009) Induction of oncogene addiction shift to NF-kappaB by camptothecin in solid tumor cells. Biochem Biophys Res Commun 390:60–64CrossRefPubMedGoogle Scholar
  27. 27.
    Capone ML, Tacconelli S, Sciulli MG, Patrignani P (2003) Clinical pharmacology of selective COX-2 inhibitors. Int J Immunopathol Pharmacol 16:49–58CrossRefPubMedGoogle Scholar
  28. 28.
    Patrignani P, Patrono C (2015) Cyclooxygenase inhibitors: from pharmacology to clinical read-outs. Biochim Biophys Acta 1851:422–432CrossRefPubMedGoogle Scholar
  29. 29.
    Pordanjani SM, Hosseinimehr SJ (2016) The Role of NF-kB Inhibitors in cell response to radiation. Curr Med Chem 23:3951–3963CrossRefPubMedGoogle Scholar
  30. 30.
    Pilones KA, Vanpouille-Box C, Demaria S (2015) Combination of radiotherapy and immune checkpoint inhibitors. Semin Radiat Oncol 25:28–33CrossRefPubMedGoogle Scholar
  31. 31.
    Esposito A, Criscitiello C, Curigliano G (2015) Immune checkpoint inhibitors with radiotherapy and locoregional treatment: synergism and potential clinical implications. Curr Opin Oncol 27:445–451CrossRefPubMedGoogle Scholar
  32. 32.
    Derer A, Frey B, Fietkau R, Gaipl US (2016) Immune-modulating properties of ionizing radiation: rationale for the treatment of cancer by combination radiotherapy and immune checkpoint inhibitors. Cancer Immunol Immunother 65:779–786CrossRefPubMedGoogle Scholar
  33. 33.
    Demaria S, Coleman CN, Formenti SC (2016) Radiotherapy: changing the game in immunotherapy. Trends Cancer 2:286–294CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2017

Authors and Affiliations

  1. 1.College of Veterinary MedicineTuskegee UniversityTuskegeeUSA
  2. 2.University of Texas El PasoEl PasoUSA
  3. 3.Wallace Tumor InstituteUniversity of Alabama BirminghamBirminghamUSA

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