Synergistic Effect of Subtoxic-dose Cisplatin and TRAIL to Mediate Apoptosis by Down-regulating Decoy Receptor 2 and Up-regulating Caspase-8, Caspase-9 and Bax Expression on NCI-H460 and A549 Cells

Document Type: Original Article


1 1Department of Medical Oncology, Cancer Center of West China Hospital, Sichuan University, Chengdu, Sichuan Province, China

2 Department of Oncology, Hebei General Hospital, Shijiazhuang, Hebei Province, China

3 2Department of Pathophysiology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu, Sichuan Province, China


Objective(s): Although tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) can selectively induce apoptosis in tumor cells, more than half of tumors including non-small cell lung cancer (NSCLC) exhibit TRAIL-resistance. The purpose of this study was to determine whether subtoxic-dose cisplatin and TRAIL could synergistically enhance apoptosis on NSCLC cells and investigate its underlying mechanisms.
Materials and Methods:NCI-H460 and A549 cells were treated with TRAIL alone, cisplatin alone or combination treatment in this study. The cytotoxicity was evaluated according to Sulforhodamine B assay, and apoptosis was examined using Hoechst 33342 staining and flow cytometry. The mRNA and protein levels of TRAIL receptors and apoptotic proteins including caspase-8, caspase-9, Bcl-2 and Bax were determined by RT-PCR and Western blotting, respectively.
Results:Our results showed that NCI-H460 cells were sensitive to TRAIL, whereas A549 cells were resistant. However, subtoxic-dose cisplatin could enhance the both cells to TRAIL-mediated cell proliferation inhibition and apoptosis. The underlying mechanisms might be associated with the down-regulation of DcR2 and up-regulation of Caspase-8, Caspase-9 and Bax.
Conclusion:Subtoxic-dose cisplatin could enhance both TRAIL- sensitive and TRAIL- resistant NSCLC cells to TRAIL-mediated apoptosis. These findings motivated further studies to evaluate such a combinatory therapeutic strategy against NSCLC in the animal models.


  1.  Charpidou A, Syrigos K. Developments in the treatment of non-small cell lung cancer. Anticancer Res 2007; 27: 2823-2827.
  2.  Bunn PA Jr, Vokes EE, Langer CJ, Schiller JH. An update on North American  randomized studies in non-small cell lung cancer. Semin Oncol 1998; 25: 2-10.
  3.  Newsom-Davis T, Prieske S, Walczak H. Is TRAIL the holy grail of cancer therapy? Apoptosis 2009; 14(4):607-623.
  4.  Ashkenazi A. Targeting death and decoy receptors of the tumour-necrosis factor superfamily. Nat Rev Cancer 2002; 2: 420-430.
  5.  Wang S. The promise of cancer therapeutics targeting the TNF-related apoptosis-inducing ligand and TRAIL receptor pathway. Oncogene 2008; 27: 6207-6215.
  6.  Picarda G, Trichet V, Teletchea S, Heymann D, Redini F. TRAIL receptor signaling and therapeutic option in bone tumors: the trap of the bone microenvironment. Am J Cancer Res 2012; 2(1):45-64.
  7.  Sheridan JP, Marsters SA, Pitti RM, Gurney A, Skubatch M, Baldwin D, et al. Control of TRAIL-induced apoptosis by a family of signaling and decoy receptors. Science 1997; 277: 818-821.
  8.  Mousavi SH, Tayarani-Najaran Z, Hersey P. Apoptosis: from signalling pathways to therapeutic tools. Iranian J Basic Med Sci 2008; 11(3):121-142.
  9.  Mahalingam D, Szegezdi E, Keane M, de Jong S, Samali A. TRAIL receptor signalling and modulation: Are we on the right TRAIL. Cancer Treat Rev 2009; 35: 280-288.
10. Brady HJ, Gil-Gomez G. Bax. The pro-apoptotic Bcl-2 family member, Bax. Int J Biochem Cell Biol 1998; 30: 647-650.

11. Sanlioglu AD. Karacay B, Koksal IT, Griffith TS, Sanlioglu S. DcR2 (TRAIL-R4) siRNA and adenovirus delivery of TRAIL (Ad5hTRAIL) break down in vitro tumorigenic potential of prostate carcinoma cells. Cancer Gene Ther 2007; 14: 976-984.

12. Vindrieux D, Reveiller M, Chantepie J, Yakoub S, Deschildre C, Ruffion A, et al. Down-regulation of DcR2 sensitizes androgen-dependent prostate cancer LNCaP cells to TRAIL-induced apoptosis. Cancer Cell Int 2011; 11: 42-55.

13. LeBlanc H, Lawrence D, Varfolomeev E, Totpal K, Morlan J, Schow P, et al. Tumor-cell resistance to death receptor-induced apoptosis through mutational inactivation of the proapoptotic Bcl-2 homolog Bax. Nat Med 2002; 8: 274-281.

14. Kim EH, Yoon MJ, Kim SU, Kwon TK, Sohn S, ChoiKS. Arsenic trioxide sensitizes human glioma cells, but not normal astrocytes, to TRAIL-induced apoptosis via CCAAT/enhancer-binding protein homologous protein-dependent DR5 up-regulation. Cancer Res 2008; 68: 266-275.

15. Pai SI, Wu GS, Ozoren N, Wu L, Jen J, Sidransky D, et al. Rare loss-of-function mutation of a death receptor gene in head and neck cancer. Cancer Res 1998; 58: 3513-3518.

16. Teitz T, Wei T, Valentine MB, Vanin EF, Grenet J, Valentine VA, et al. Caspase 8 is deleted or silenced preferentially in childhood neuroblastomas with amplification of MYCN. Nat Med 2000; 6: 529-535.

17. Fulda S, Meyer E, Debatin KM. Inhibition of TRAIL-induced apoptosis by Bcl-2 overexpression. Oncogene 2002; 21: 2283-2294.

18. Yoo J, Park SS, Lee YJ. Pretreatment of docetaxel enhances TRAIL-mediated apoptosis in prostate cancer cells. J Cell Biochem 2008; 104: 1636-1646.

19. Lacour S, Hammann A, Wotawa A, Corcos L, Solary E, Dimanche-Boitrel MT. Anticancer agents sensitize tumor cells to TRAIL-mediated activation and apoptosis. Cancer Res 2001; 61: 1645-1651.

20. Hotta T, Suzuki H, Nagai S, Yamamoto K, Imakiire A, Takada E, et al. Chemotherapeutic agents sensitize sarcoma cell lines to tumor necrosis factor-related apoptosis-inducing ligand-induced caspase-8 activation, apoptosis and loss of mitochondrial membrane potential. J Orthop Res 2003; 21: 949-957.

21. Huang Y, Liu Y, Zhou J, Xu N, Li B, Wu G, et al. A randomized, controlled, multicenter clinical trial comparing pemetrexed/cisplatin and gemcitabine/ cisplatin as first-line treatment for advanced nonsquamous non-small cell lung cancer. Zhongguo Fei Ai Za Zhi 2012; 15(10):576-582.

22. Inal A, Kaplan MA, Kucukoner M, Karakus A, Isikdogan A. Prognostic factors in elderly patients with advanced non-small cell lung cancer treated with first-line cisplatin-based chemotherapy: a retrospective analysis of single institution. J BUON 2012; 17(3):533-536.

23. Karimi G, Khoei A, Omidi A, Kalantari M, Babaei J, Taghiabadi E, et al. Protective effect of aqueous and ethanolic extracts of Portulaca oleracea against cisplatin induced nephrotoxicity. Iran J Basic Med Sci 2010; 13:31-35.

24. Lee KW, Jeong JY, Lim BJ, Chang Y-K, Lee S-J, Na K-R, et al. Sildenafil attenuates renal injury in an experimental model of rat cisplatin-induced nephrotoxicity. Toxicology 2009; 257:137-143.

25. Kuhlmann MK, Horsch E, Burkhardt G, Wagner M, Kohler H. Reduction of cisplatin toxicity in cultured renal tubular cells by the bioflavonoid quercetin. Arch Toxicol 1998; 72:536-540.

26. Woods DC, Alvarez C, Johnson AL. Cisplatin-mediated sensitivity to TRAIL-induced cell death in human granulosa tumor cells. Gynecol Oncol 2008; 108: 632-640.

27. Xu L, Yin S, Reddy KB. Enhanced anticancer effect of the combination of cisplatin and TRAIL in triple-negative breast tumor cells. Mol Cancer Ther 2011; 10: 550-557.

28. Skehan P, Storeng R, Scudiero D, Monks A, McMahon J, Vistica D, et al. New colorimetric cytotoxicity assay for anticancer-drug screening. J Natl Cancer Inst 1990; 82: 1107-1112.

29. Cao JY, Liu L, Chen SP, Zhang X, Mi YJ, Liu ZG, et al. Prognostic significance and therapeutic implications of centromere protein F expression in human nasopharyngeal carcinoma. Mol Cancer 2010; 9: 237-249.

30. Nelson K, Walsh D, Sheehan F. Cancer and chemotherapy-related upper gastrointestinal symptoms: the role of abnormal gastric motor function and its evaluation in cancer patients. Support Care Cancer 2002; 10: 455-461.

31. Schnell FM. Chemotherapy-induced nausea and vomiting: the importance of acute antiemetic control. Oncologist 2003; 8: 187-198.

32. Dubey S, Schiller JH. Chemotherapy for advanced non-small cell lung cancer. Hematol Oncol Clin North Am 2004; 18: 101-114.

33. Xie ZH, Quan MF, Liu F, Cao JG, Zhang JS. 5-allyl-7-gen- difluoromethoxychrysin enhances TRAIL-induced apoptosis in human lung carcinoma A549 cells. BMC Cancer 2011; 11:322-332.

34. Jang JY, JeonYK, Choi Y, Kim CW. Short-hairpin RNA-induced suppression of adenine nucleotide translocase-2 in breast cancer cells restores their susceptibility to TRAIL-induced apoptosis by activating JNK and modulating TRAIL receptor expression. Mol Cancer 2010; 9: 262-273.

35. Pan G, Ni J, Wei YF, Yu G, Gentz R, Dixit VM. An antagonist decoy receptor and a death domain-containing receptor for TRAIL. Science 1997; 277: 815-818.

36. Balsas P, Lopez-Royuela N, Galan-Malo P, Anel A, Marzo I, Naval J. Cooperation between Apo2L/TRAIL and bortezomib in multiple myeloma apoptosis. Biochem Pharmacol 2009; 77: 804-812.