Effect of Embelin on TRAIL Receptor 2 mAb-induced Apoptosis of TRAIL-resistant A549 Non-small Cell Lung Cancer Cells

  • Jiang, Lei (Department of Pathology, Beijing Chao-Yang Hospital, Capital Medical University) ;
  • Hao, Jin-Li (Department of Pathology, Beijing Chao-Yang Hospital, Capital Medical University) ;
  • Jin, Mu-Lan (Department of Pathology, Beijing Chao-Yang Hospital, Capital Medical University) ;
  • Zhang, Yun-Gang (Department of Pathology, Beijing Chao-Yang Hospital, Capital Medical University) ;
  • Wei, Ping (Department of Pathology, Beijing Chao-Yang Hospital, Capital Medical University)
  • Published : 2013.10.30


Introduction: Some non-small cell lung cancer (NSCLC) tumor cells are insensitive to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) -based therapy. This study was conducted to examine the effect of embelin on the sensitivity of the A549 NSCLC cell line to TRAIL receptor2 (TRAILR2) monoclonal antibodies and to investigate the potential mechanisms. Materials and Methods: A549 cells were treated with embelin, TRAILR2 mAb or a combination of both. Cell viability was measured using ATPlite assay and apoptosis rates were determined by flow cytometry with AnnexinV-FITC and propidium iodide staining, with the expression levels of proteins analyzed by Western blotting. Results: The cell survival rate of separate treatments with 100 ng/ml TRAILR2 antibody or 25 uM embelin were $81.5{\pm}1.57%$ and $61.7{\pm}2.84%$, respectively. Their combined use markedly decreased cell viability in A549 cells to $28.1{\pm}1.97%$ (P<0.05). The general caspase inhibitor Z-VAD-FMK could inhibit the embelin-enhanced sensitivity of A549 cells to TRAILR2 mAb ($75.97{\pm}3.17%$)(P<0.05). Both flow cytometry and cell morphological analysis showed that embelin was able to increase TRAIL-induced apoptosis in A549 cells. Combined treatment with embelin and TRAILR2 mAb augmented the activation of initiator caspases and effector caspase. In addition, A549 cells showed increasing levels of TRAILR2 protein and decreasing levels of Bcl-2, survivin and c-FLIP following the treatment with embelin+TRAILR2 mAb. Conclusions: Embelin could enhance TRAIL-induced apoptosis in A549 cells. The synergistic effect of the combination treatment might be due to modulation of multiple components in the TRAIL receptor-mediated apoptotic signaling pathway, including TRAILR2, XIAP, survivin, Bcl-2 and c-FLIP.


TRAIL;embelin;lung cancer;apoptosis


  1. Cooper WA, Kohonen-Corish MR, Zhuang L, et al (2008). Role and prognostic significance of tumor necrosis factor-related apoptosis-inducing ligand death receptor DR5 in nonsmall-cell lung cancer and precursor lesions. Cancer, 113, 135-42.
  2. Cheng YJ, Jiang HS, Hsu SL, et al (2010). XIAP-mediated protection of H460 lung cancer cells against cisplatin. Eur J Pharmacol, 627, 75-84.
  3. Chitra M, Sukumar E, Suja V, Devi CS (1994). Antitumor, antiinflammatory and analgesic property of embelin, a plant product. Chemotherapy, 40, 109-13.
  4. Chou TC (2008). Preclinical versus clinical drug combination studies. Leuk Lymphoma, 49, 2059-80.
  5. Dai Y, Qiao L, Chan KW, et al (2009). Peroxisome proliferatoractivated receptor-gamma contributes to the inhibitory effects of Embelin on colon carcinogenesis. Cancer Res, 69, 4776-83.
  6. De Petris L, Crino L, Scagliotti GV, et al (2006). Treatment of advanced non-small cell lung cancer. Ann Oncol, 17, ii36-41.
  7. Duiker EW, Mom CH, de Jong S, et al (2006). The clinical trail of TRAIL. Eur J Cancer, 42, 2233-40.
  8. Fesik SW (2005). Promoting apoptosis as a strategy for cancer drug discovery. Nat Rev Cancer, 5, 876-85.
  9. Field JK, Duffy SW (2008). Lung cancer screening: the way forward. Br J Cancer, 99, 557-62.
  10. Gross A, McDonnell JM, Korsmeyer SJ (1999). BCL-2 family members and the mitochondria in apoptosis. Genes Dev, 13, 1899-911.
  11. Holcik M, Gibson H, Korneluk RG (2001). XIAP: apoptotic brake and promising therapeutic target. Apoptosis, 6, 253-61.
  12. Kim K, Fisher MJ, Xu SQ, el-Deiry WS (2000). Molecular determinants of response to TRAIL in killing of normal and cancer cells. Clin Cancer Res, 6, 335-46.
  13. Ihde DC, Minna JD (1991). Non-small cell lung cancer. Part II: Treatment. Curr Probl Cancer, 15, 105-54.
  14. Jo M, Kim TH, Seol DW, et al (2000). Apoptosis induced in normal human hepatocytes by tumor necrosis factor-related apoptosis-inducing ligand. Nat Med, 6, 564-7.
  15. Kaminskyy VO, Surova OV, Piskunova T, et al (2013). Upregulation of c-FLIP-short in response to TRAIL promotes survival of NSCLC cells, which could be suppressed by inhibition of Ca2+/calmodulin signaling. Cell Death Dis, 4, e522.
  16. Koschny R, Ganten TM, Sykora J, et al (2007). TRAIL/bortezomib cotreatment is potentially hepatotoxic but induces cancer-specific apoptosis within a therapeutic window. Hepatology, 45, 649-58.
  17. Lan Y, Liu X, Zhang R, et al (2013). Lithium enhances TRAIL-induced apoptosis in human lung carcinoma A549 cells. Biometals, 26, 241-54.
  18. Li XQ, Ke XZ, Wang YM (2012). Treatment of malignant melanoma by downregulation of XIAP and overexpression of TRAIL with a conditionally replicating oncolytic adenovirus. Asian Pac J Cancer Prev, 13, 1471-6.
  19. Luster TA, Carrell JA, McCormick K, et al (2009). Mapatumumab and lexatumumab induce apoptosis in TRAIL-R1 and TRAIL-R2 antibody-resistant NSCLC cell lines when treated in combination with bortezomib. Mol Cancer Ther, 8, 292-302.
  20. Mahalingam D, Szegezdi E, Keane M, et al (2009). TRAIL receptor signalling and modulation: Are we on the right TRAIL? Cancer Treat Rev, 35, 280-8.
  21. Siegelin MD, Gaiser T, Siegelin Y (2009). The XIAP inhibitor Embelin enhances TRAIL-mediated apoptosis in malignant glioma cells by down-regulation of the short isoform of FLIP. Neurochem Int, 55, 423-30.
  22. Mori T, Doi R, Kida A, et al (2007). Effect of the XIAP inhibitor Embelin on TRAIL-induced apoptosis of pancreatic cancer cells. J Surg Res, 142, 281-6.
  23. Nikolovska-Coleska Z, Xu L, Hu Z, et al (2004). Discovery of embelin as a cell-permeable, small-molecular weight inhibitor of XIAP through structure-based computational screening of a traditional herbal medicine three-dimensional structure database. J Med Chem, 47, 2430-40.
  24. Pore MM, Hiltermann TJ, Kruyt FA (2010). Targeting apoptosis pathways in lung cancer. Cancer Lett, 332, 359-68.
  25. Spierings DC, de Vries EG, Timens W, et al (2003). Expression of TRAIL and TRAIL death receptors in stage III non-small cell lung cancer tumors. Clin Cancer Res, 9, 3397-405.
  26. Sprick MR, Weigand MA, Rieser E, et al (2000). FADD/MORT1 and caspase-8 are recruited to TRAIL receptors 1 and 2 and are essential for apoptosis mediated by TRAIL receptor 2. Immunity, 12, 599-609.
  27. Stegehuis JH, de Wilt LH, de Vries EG, et al (2009). TRAIL receptor targeting therapies for non-small cell lung cancer: current status and perspectives. Drug Resist Updat, 13, 2-15.
  28. Voortman J, Resende TP, Abou El Hassan MA, et al (2007). TRAIL therapy in non-small cell lung cancer cells: sensitization to death receptor-mediated apoptosis by proteasome inhibitor bortezomib. Mol Cancer Ther, 6, 2103-12.
  29. Zhang L, Fang B (2005). Mechanisms of resistance to TRAIL-induced apoptosis in cancer. Cancer Gene Ther, 12, 228-37.

Cited by

  1. Dealing Naturally with Stumbling Blocks on Highways and Byways of TRAIL Induced Signaling vol.15, pp.19, 2014,
  2. Drugs from Marine Sources: Modulation of TRAIL Induced Apoptosis in Cancer Cells vol.15, pp.20, 2014,
  3. Quercetin-induced autophagy flux enhances TRAIL-mediated tumor cell death vol.34, pp.1, 2015,
  4. The XIAP inhibitor Embelin enhances TRAIL-induced apoptosis in human leukemia cells by DR4 and DR5 upregulation vol.36, pp.2, 2015,
  5. Embelin induces apoptosis of human gastric carcinoma through inhibition of p38 MAPK and NF-κB signaling pathways vol.14, pp.1, 2016,
  6. Effect of processing techniques on new poly(ε-caprolactone)-embelin microparticles of biomedical interest pp.07306679, 2017,
  7. The Application of Embelin for Cancer Prevention and Therapy vol.23, pp.3, 2018,
  8. Embelin: a benzoquinone possesses therapeutic potential for the treatment of human cancer vol.10, pp.8, 2018,