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Synergistic antitumor activity of TRAIL combined with chemotherapeutic agents in A549 cell lines in vitro and in vivo

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Abstract

Purpose

To investigate the synergistic cytotoxicity of TRAIL in combination with chemotherapeutic agents in A549 cell lines, we systematically evaluated the cytotoxicity of TRAIL alone and TRAIL in combination with cisplatin, paclitaxel (Taxol) or actinomycin D in A549 cell lines in vitro and in vivo, and whether the sensitivity was correlated with the expression level of TRAIL receptors.

Methods

We investigated the cytotoxicity of TRAIL alone and the synergistic antitumor effects of TRAIL in combination with chemotherapeutic agents in A549 cells by crystal violet staining and FACS in vitro. The expression levels of DR4, DR5, DcR1 and DcR2 were measured in TRAIL-treated and chemotherapeutic agent-treated A549 cells by Western blotting. The growth inhibition of tumors was evaluated in terms of incidence, volume and weight in a A549-implanted nude mice model.

Results

Chemotherapeutic agents cisplatin (5.56 μg/ml), Taxol (10 and 30 μg/ml) or actinomycin D (9.26, 83.3 and 750 ng/ml) augmented the cytotoxicity of TRAIL in A549 cell lines within a range of concentrations of TRAIL (1.98–160 ng/ml) in vitro. The expression levels of DR4 and DR5 were not significantly different and the expression of DcR2 was slightly downregulated, but the expression of DcR1 was not detected in non-treated, TRAIL-treated and chemotherapeutic agent-treated A549 cells. The rates of tumor inhibition following treatment with TRAIL alone (15 mg/kg per day, daily for 10 days) and TRAIL/cisplatin (15 mg/kg per day TRAIL, daily for 10 days; 1.5 mg/kg per day cisplatin, daily for 10 days with 7-day intervals) were 28.3% and 76.8% by tumor weight (P<0.05 for TRAIL alone versus control, P<0.05 for TRAIL/cisplatin versus cisplatin alone and TRAIL alone) on day 65 in vivo.

Conclusion

TRAIL in combination with chemotherapeutic agents cisplatin, Taxol or actinomycin D exerted synergistic antitumor effects in A549 cell lines in vitro and TRAIL/cisplatin demonstrated synergistic antitumor effects in vivo. The expression levels of TRAIL receptors suggested that the synergistic effects of TRAIL in combination with chemotherapeutic agents are not at the receptor level in A549 cell lines.

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Abbreviations

TNF:

Tumor necrosis factor

TRAIL:

TNF-related apoptosis-inducing ligand

NSCLC:

Non-small cell lung cancer

DR:

Death receptor

DcR:

Decoy receptor

IB:

Inclusion body

FACS:

Fluorescence activated cell sorter

PI:

Propidium iodide

IR:

Rate of inhibition

PMSF:

Phenyl methyl sulfonyl fluoride

References

  1. Jemal A, Thomas A, Murray T, Thun M (2002) Cancer statistics, 2002. CA Cancer J Clin 52:23–47

    PubMed  Google Scholar 

  2. Peter ME, Krammer PH (1998) Mechanisms of CD95 (Apo-1/Fas)-mediated apoptosis. Curr Opin Immunol 10:545–551

    Article  CAS  PubMed  Google Scholar 

  3. Wiley SR, Schooley K, Smolak PJ, Din WS, Hunag CP, Nicholl JK, Sutherland GR, Smith TD, Rauch C, Smith CA (1995) Identification and characterization of a new member of the TNF family that induces apoptosis. Immunity 3:673–682

    Article  CAS  PubMed  Google Scholar 

  4. Pitti RM, Marsters SA, Ruppert S, Donahue CJ, Moore A, Ashkenazi A (1996) Induction of apoptosis by Apo-2 ligand, a new member of the tumor necrosis factor cytokine family. J Biol Chem 271:12687–12690

    Article  CAS  PubMed  Google Scholar 

  5. Ashkenazi A, Pai RC, Song S, Leung S, Lawrence DA, Marsters SA, Blackie C, Chang L, McMurtrey AE, Hebert A, DeForge L, Koumenis IL, Lewis D, Harris L, Bussiere J, Koeppen H, Shahrohk Z, Schwall RH (1999) Safety and antitumor activity of recombinant soluble Apo2 ligand. J Clin Invest 104:155–162

    CAS  PubMed  Google Scholar 

  6. Walczak H, Miller RE, Ariail K, Gliniak B, Griffith TS, Kubin K, Chin W, Jones J, Woodward A, Le T, Smith C, Smolak P, Goodwin RG, Rauch CT, Schun JCL, Lynch D (1999) Tumoricidal activity of tumor necrosis factor-related apoptosis-inducing ligand in vivo. Nat Med 5:157–163

    Article  CAS  PubMed  Google Scholar 

  7. Pan G, O’Rourke K, Chinnaiyan AM, Gentz R, Ebner R, Ni J, Dixit VM (1997) The receptor for the cytotoxic ligand TRAIL. Science 276:111–113

    Article  CAS  PubMed  Google Scholar 

  8. Screaton GR, Mongkolsapaya J, Xu XN, Cowper AE, McMichael AJ, Bell JI (1997) TRICK2, a new alternatively spliced receptor that transduces the cytotoxic signal from TRAIL. Curr Biol 7:693–696

    Article  CAS  PubMed  Google Scholar 

  9. Sheridan JP, Marsters SA, Pitti RM, Gurney A, Skubatch M, Baldwin D, Ramakrishnan L, Gray CL, Baker K, Wood WI, Goddard AD, Godowski P, Ashkenazi A (1997) Control of TRAIL-induced apoptosis by a family of signaling and decoy receptors. Science 277:818–821

    Article  PubMed  Google Scholar 

  10. Ashkenazi A, Dixit VM (1998) Death receptor: signaling and modulation. Science 281:818–821

    Article  PubMed  Google Scholar 

  11. Gura T (1997) How TRAIL kills cancer cells, but not normal cells. Science 277:768

    Article  CAS  PubMed  Google Scholar 

  12. Griffith TS, Ranth CT, Smolak PJ, Waugh JY, Boiani N, Lynch DH, Smith CA, Goodwin RG, Kubin MZ (1999) Function analysis of TRAIL receptor using monoclonal antibody. J Immunol 162:2597–2605

    CAS  PubMed  Google Scholar 

  13. Kim K, Fisher MA, Xu SQ, EI-Deiry WS (2000) Molecular determinants of response to TRAIL in killing of normal and cancer cells. Clin Cancer Res 6:335–346

    CAS  PubMed  Google Scholar 

  14. Bodmer JL, Holler N, Reynad S, Vinciguerra P, Schneider P, Juo P, Blenis J, Tschopp J (2000) TRAIL receptor-2 signals apoptosis through FADD and caspase-8. Nat Cell Biol 2:241–243

    Article  CAS  PubMed  Google Scholar 

  15. Suliman A, Lam A, Datta R, Srivastava RK (2001) Intracellular mechanisms of TRAIL: apoptosis through mitochondrial-dependent and -independent pathways. Oncogene 20:2122–2133

    Article  CAS  PubMed  Google Scholar 

  16. Chinnaiyan AM, Prasad U, Shankar S, Hamstra DA, Shanaiah M, Chenevert TL, Ross BD, Rehemtulla A (2000) Combined effect of tumor necrosis factory-related apoptosis-inducing ligand and ionizing radiation in breast cancer therapy. Proc Natl Acad Sci U S A 97:1754–1759

    Article  CAS  PubMed  Google Scholar 

  17. Kelley SK, Harris LA, Xie D, Deforge L, Totpal K, Bussiere J, Fox JA (2001) Preclinical studies to predict the disposition of Apo2L/tumor necrosis factor-related apoptosis-inducing ligand in humans: characterization of in vivo efficacy, pharmacokinetics, and safety. J Pharmacol Exp Ther 299:31–38

    CAS  PubMed  Google Scholar 

  18. Hymowitz SG, O’Connell MP, Ultsch MH, Hurst A, Totpal K, Ashkenazi A, de Vos AM, Kelley RF (2000) A unique zinc-binding site revealed by a high-resolution X-ray structure of homotrimeric Apo2L/TRAIL. Biochemistry 39:633–640

    Article  CAS  PubMed  Google Scholar 

  19. Bodmer JL, Meier P, Tschopp J, Schneider P (2000) Cysteine 230 is essential for the structure and activity of the cytotoxic ligand TRAIL. J Biol Chem 275:20632–20637

    Article  CAS  PubMed  Google Scholar 

  20. Sherwood SW, Schimke RT (1995) Cell cycle analysis of apoptosis using flow cytometry. Methods Cell Biol 46:77–97

    CAS  PubMed  Google Scholar 

  21. Jin ZJ (1980) Linear transformation of Michaelis–Menton equation by Lineweaver–Burk’s method. Acta Pharmacol Sin 1:3–7

    CAS  Google Scholar 

  22. Kotoh T, Kumar D, Masunaga R, Tabara H, Tachibana M, Kubota H, Hohno H, Nagasue N (1999) Antiangiogenic therapy of human esophageal cancers with thalidomide in nude mice. Surgery 125:536–544

    Article  CAS  PubMed  Google Scholar 

  23. Jean Marie BR, Shyamal KM (2002) Antitumorigenic evaluation of thalidomide alone and in combination with cisplatin in DBA2/J mice. J Biomed Biotechnol 2:7–13

    Article  PubMed  Google Scholar 

  24. Havell EA, Fires W, North RJ (1988) The antitumor function of tumor necrosis factor (TNF). I. Therapeutic action of TNF against an established murine sarcoma is indirect, immunologically dependent, and limited by severe toxicity. J Exp Med 167:1067–1085

    Article  CAS  PubMed  Google Scholar 

  25. Ogasawara J, Wantanbe-Fuknnaga R, Adachi M (1993) Lethal effect of the anti-Fas antibody in mice. Nature 364:806–809

    Article  CAS  PubMed  Google Scholar 

  26. Frese S, Brunmer T, Gugger M, Uduehi A, Schmid RA (2002) Enhancement of Apo2L/TRAIL (tumor necrosis factor-related apoptosis-inducing ligand)-induced in non-small cell lung cancer cell lines by chemotherapeutic agents without correlation to the expression level of cellular protease caspase-8 inhibitory protein. J Thorac Cardiovasc Surg 123:168–174

    Article  CAS  PubMed  Google Scholar 

  27. Mizutani Y, Yoshida O, Tsuneharn M, Bonavida B (1999) Synergistic cytotoxicity and apoptosis by Apo-2 ligand and adriamycin against bladder cancer cells. Clin Cancer Res 5:2605–2612

    CAS  PubMed  Google Scholar 

  28. Keane MM, Ettenberg SA, Nau MM, Russell EK, Lipkowitz S (1999) Chemotherapeutic augments TRAIL-induced apoptosis in breast cell lines. Cancer Res 59:734–741

    CAS  PubMed  Google Scholar 

  29. Munshi AT, Meyn MR (2002) Chemotherapeutic agents enhance TRAIL-induced apoptosis in prostate cancer cells. Cancer Chemother Pharmacol 50:46–52

    Article  CAS  PubMed  Google Scholar 

  30. Griffith TS, Chin WA, Jackson GC, Lynch DH, Kubin MZ (1998) Intracellular regulation of TRAIL-induced apoptosis in human melanoma cells. J Immunol 161:2833–2840

    CAS  PubMed  Google Scholar 

  31. Nagane M, Pan G, Weddle JJ, Dixit VM, Cavenee WK, Huang HJS (2000) Increased death receptor 5 expression by chemotherapeutic agents in human gliomas causes synergistic cytotoxicity with tumor necrosis factor-related apoptosis-inducing ligand in vitro and in vivo. Cancer Res 60:847–853

    CAS  PubMed  Google Scholar 

  32. Arizono A, Yoshikawa H, Naganuma H, Hamada Y, Nakajima Y, Tasaka K (2003) A mechanism of resistance to TRAIL/Apo2L-induced apoptosis of newly established glioma cell line and sensitization to TRAIL by genotoxic agents. Br J Cancer 88:298–306

    Article  CAS  PubMed  Google Scholar 

  33. Bouralexi S, Findlay DM, Atkins GJ, Labrinidis A, Hay S, Evdokiou (2003) Progressive resistance of BTK osteosarcoma cells to Apo2L/TRAIL-induced apoptosis is mediated by acquisition of DcR2/TRAIL-R4 expression: resensitisation with chemotherapy. Br J Cancer 89:206–214

    Article  PubMed  Google Scholar 

  34. Geelen CMM, Vries EGE, Le TKP, Weeghel RP, Jong SD (2003) Differential modulation of the TRAIL receptors and the CD95 receptor in colon carcinoma cell lines. Br J Cancer 89:363–373

    Article  PubMed  Google Scholar 

  35. Gliniak B, Le T (1999) Tumor necrosis factor-related apoptosis-inducing ligand’s antitumor in vivo is enhanced by the chemotherapeutic agent CPT-11. Cancer Res 59:6153–6158

    CAS  PubMed  Google Scholar 

  36. Singh TR, Shankar S, Chen X, Asim M, Srivastava RK (2003) Synergistic interactions of chemotherapeutic drugs and tumor necrosis factor-related apoptosis-inducing ligand/Apo-2 ligand on apoptosis and on regression of breast carcinoma in vivo. Cancer Res 63:5390–5400

    CAS  PubMed  Google Scholar 

  37. Ray S, Almasan A (2003) Apoptosis induction in prostate cancer cells and xenografts by combined treatment with Apo2 ligand/tumor necrosis factor-related apoptosis-inducing ligand and CPT-11. Cancer Res 63:4713–4723

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by grants from the Tenth Five-Year Research Project of Anhui Province. We thank Dr. Zhimin Zhai for FACS analysis, the Central Laboratory of the Provincial Hospital of Anhui and Dr. Plascal Schneider (Institute of Biochemistry, University of Lausanne, Switzerland) for Flag-TRAIL and Anti-Flag M2 antibody,.

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Authors and Affiliations

  1. Institute of Clinical Pharmacology, Anhui Medical University, Hefei, 230032, People’s Republic of China

    Qing-Lin Fan & Wei Wei

  2. Molecular Biology Lab, Biological Institute of Anhui Province, Hefei, 230088, People’s Republic of China

    Qing-Lin Fan & Li-Hua Song

  3. Molecular Biology Lab, Anhui Anke Biotechnology Co. Ltd, Hefei, 230088, People’s Republic of China

    Qing-Lin Fan, Wen-Yi Zou & Li-Hua Song

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  1. Qing-Lin Fan
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  2. Wen-Yi Zou
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Correspondence to Li-Hua Song or Wei Wei.

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Fan, QL., Zou, WY., Song, LH. et al. Synergistic antitumor activity of TRAIL combined with chemotherapeutic agents in A549 cell lines in vitro and in vivo. Cancer Chemother Pharmacol 55, 189–196 (2005). https://doi.org/10.1007/s00280-004-0867-1

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  • DOI: https://doi.org/10.1007/s00280-004-0867-1

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