Quercetin Potentiates TRAIL-induced Apoptosis in Human Colon KM12 Cells

사람 대장암 KMl2세포에서 quercetin 의한 TRAIL이 유도하는 세포사멸의 증가

  • Park, Jun-Ik (Department of Biochemistry, Pusan National University School of Medicine) ;
  • Kim, Hak-Bong (Department of Biochemistry, Pusan National University School of Medicine) ;
  • Kim, Mi-Ju (Department of Biochemistry, Pusan National University School of Medicine) ;
  • Lee, Jae-Won (Department of Biochemistry, Pusan National University School of Medicine) ;
  • Bae, Jae-Ho (Department of Biochemistry, Pusan National University School of Medicine) ;
  • Park, Soo-Jung (Division of Intractable Diseases, Center for Biomedical Sciences, National Institute of Health) ;
  • Kim, Dong-Wan (Department of Microbiology, College of Natural Sciences, Chang Won National University) ;
  • Kang, Chi-Dug (Department of Biochemistry, Pusan National University School of Medicine) ;
  • Kim, Sun-Hee (Department of Biochemistry, Pusan National University School of Medicine)
  • 박준익 (부산대학교 의학전문대학원 생화학교실) ;
  • 김학봉 (부산대학교 의학전문대학원 생화학교실) ;
  • 김미주 (부산대학교 의학전문대학원 생화학교실) ;
  • 이재원 (부산대학교 의학전문대학원 생화학교실) ;
  • 배재호 (부산대학교 의학전문대학원 생화학교실) ;
  • 박수정 (국립보건원 만성질환연구부) ;
  • 김동완 (창원대학교 자연과학대 미생물학과) ;
  • 강치덕 (부산대학교 의학전문대학원 생화학교실) ;
  • 김선희 (부산대학교 의학전문대학원 생화학교실)
  • Published : 2009.09.30


Many cancer cells are sensitive to the TNF-related apoptosis-inducing ligand (TRAIL)-induced apoptosis. However, some cancer cells show either partial or complete resistance to TRAIL. Human colon carcinoma KM12 cells have been shown to be insensitive to TRAIL-induced apoptosis. To overcome TRAIL resistance in KM12 cells, we targeted key anti-apoptotic molecules involved in the modulation of TRAIL resistance in the cells, and evaluated the effects of quercetin as a TRAIL sensitizer in the cells. We found that quercetin acted in synergy with TRAIL to enhance TRAIL-induced apoptosis in KM12 cells by the down-regulation of c-FLIP and DNA-PKcs/Akt and up-regulation of death receptors (DR4/DR5), which led to the enhancement of TRAIL-mediated activation of caspases and subsequent cleavage of PARP, as well as up-regulation of Bax. These findings suggest that the DNA-PKcs/Akt signaling pathway, as well as c-FLIP, play essential roles in regulating cells in the escape from TRAIL-induced apoptosis. Based on these results, this study provides a potential application of quercetin in combination with TRAIL in the treatment of human colon cancer.


  1. Almasan, A and A Ashkenazi. 2003. Apo2LjTRAIL: apoptosis signaling, biology, and potential for cancer therapy. Cytokine Growth Factor Rev. 14, 337-348
  2. Asakuma, J., M. Sumitomo, T. Asano, T. Asano, and M. Hayakawa. 2003. Selective Akt inactivation and tumor necrosis actor-related apoptosis-inducing ligand sensitization of renal cancer cells by low concentrations of paclitaxel. Cancer Res. 63, 1365-1370
  3. Downward, J. 2004. PI 3-kinase, Akt and cell survival. Semin. Cell Dev. BioI. 15, 177-182
  4. Dragoi, A M., X. Fu, S. Ivanov, P. Zhang, 1. Sheng, D. Wu, G. C. Li, and W. M. Chu. 2005. DNA-PKcs, but not TLR9, is required for activation of Akt by CpG-DNA EMBO J. 24, 779-789
  5. Dyer, M. J., M. MacFarlane, and G. M. Cohen. 2007. Barriers to effective TRAIL-targeted therapy of malignancy. J. Clin. Oncol. 25, 4505-4506
  6. Elnemr, A, T. Ohta, A Yachie, M. Kayahara, H. Kitagawa, T. Fujimura, I. Ninomiya, S. Fushida, G. I. Nishimura, K. Shimizu, and K. Miwa. 2001. Human pancreatic cancer cells disable function of Fas receptors at several levels in Fas signal transduction pathway. Int. J. Oncol. 18, 311-316
  7. Feng, J., J. Park, P. Cron, D. Hess, and B. A Hemmings. 2004. Identification of a PKB / Akt hydrophobic motif Ser-473 kinase as DNA-dependent protein kinase. J. Biol. Chem. 279, 41189-41196
  8. Fulda, S., I. Jeremias, and K. M. Debatin. 2004. Cooperation of betulinic acid and TRAIL to induce apoptosis in tumor cells. Oncogene 23, 7611-7620
  9. Gamet-Payrastre, L., S. M. Manenti, P. Gratacap, J. Tulliez, H. Chap, and B. Payrastre. 1999. Flavonoids and the inhibition of PKC and PI 3-kinase. Gen. Pharmacol. 32, 279-286
  10. Gerhauser, C. 2008. Cancer chemopreventive potential of apples, apple juice, and apple components. Planta Med. 74, 1608-1624
  11. Gorlick R and J. R Bertino. 1999. Drug resistance in colon cancer. Semin. Oncol. 26, 606-611
  12. Izzard, R A, S. P. Jackson, and G. C. Smith. 1999. Competitive and noncompetitive inhibition of the DNA-dependent protein kinase. Cancer Res. 59, 2581-2586
  13. Kennedy, S. G., E. S. KandeL T. K. Cross, and N. Hay. 1999. Akt/Protein kinase B inhibits cell death by preventing the release of cytochrome c from mitochondria. Mol. Cell BioI. 19, 5800-5810
  14. Kim, S. H, J. HUm, D. W. Kim, B. H Kwon, D. W. Kim, B. S. Chung, and C. D. Kang. 2000. Potentiation of chemosensitivity in multidrug-resistant human leukemia CEM cells by inhibition of DNA-dependent protein kinase using wortmannin. Leuk. Res. 24, 917-925
  15. Kim, Y. H, D. H Lee, J. H Jeong, Z. S. Guo, and Y. J. Lee. 2008. Quercetin augments TRAIL-induced apoptotic death: involvement of the ERK signal transduction pathway. Biochem. Pharmacol. 75, 1946-1958
  16. Kim, Y. Hand Y. J. Lee. 2007. TRAIL apoptosis is enhanced by quercetin through Akt dephosphorylation. J. Cell Biochem. 100, 998-1009
  17. Krueger, A, S. Baumann, P. H Krammer, and S. Kirchhoff. 2001. FUCE-inhibitory proteins: regulators of death receptor-mediated apoptosis. Mol. Cell Biol. 21, 8247-8254
  18. Kruyt, F. A 2008. TRAIL and cancer therapy. Cancer Lett. 263, 14-25
  19. Kuo, P. L., Y. L. Hsu, T. C. Lin, and C. C. Lin. 2005. The antiproliferative activity of prodelphidin B-2' 3'-0-gallate from green tea leaf is through cell cycle arrest and Fas-mediated apoptotic pathway in A549 cells. Food Chem. Toxicol. 43, 315-323
  20. Kurbanov, B. M., L. F. Fecker, C. C. Geilen, W. Sterry, and J. Eberle. 2007. Resistance of melanoma cells to TRAIL does not result from upregulation of antiapoptotic proteins by NF-kappa B but is related to downregulation of initiator caspases and DR4. Oncogene 26, 3364-3377
  21. Lane, D., V. Robert, R Grondin, C. Rancourt, and A Piche. 2007. Malignant ascites protect against TRAIL-induced apoptosis by activating the PI3K/ Akt pathway in human ovarian carcinoma cells. Int. J. Cancer 21, 1227-1237
  22. Larribere, 1., M. Khaled, S. Tartare-Deckert, R Busca, F. Luciano, K. Bille, G. Valony, A Eychene, P. Auberger, J. P. Ortonne, R Ballotti, and C. Bertolotto. 2004. PI3K mediates protection against TRAIL-induced apoptosis in primary human melanocytes. Cell Death Differ. 11, 1084-1091
  23. Lee, S. H, H S. Kim, S. Y. Kim, Y. S. Lee, W. S. Park S. H Kim, J. Y. Lee, and J. Y. Nam. 2003. Increased expression of FUP, an inhibitor of Fas-mediated apoptosis, in stomach cancer. APMIS 111, 309-314
  24. Merino, D., N. Lalaoui, A Morizot, E. Solary, and O. Micheau. 2007. TRAIL in cancer therapy: present and future challenges. Expert Opin. Ther. Targets 11, 1299-1314
  25. Micheau, 0., M. Thome, P. Schneider, N. Holler, J. Tschopp, D. W. Nicholson, C. Briand, and M. G. Grutter. 2002. The long form of FUP is an activator of caspase-8 at the Fas death-inducing signaling complex. J. Biol. Chem. 277, 45162-45171
  26. Nam, S. Y., G. A Jung, G. C. Hur, H. Y. Chung, W. H. Kim, D. W. SeoL and B. 1. Lee. 2003. Upregulation of FUP(S) by Akt, a possible inhibition mechanism of TRAIL-induced apoptosis in human gastric cancers. Cancer Sci. 94, 1066-1073
  27. Palacios, C., R Yerbes, and A Lopez-Rivas. 2006. Flavopiridol induces cellular FUCE-inhibitory protein degradation by the proteasome and promotes TRAIL-induced early signaling and apoptosis in breast tumor cells. Cancer Res. 66, 8858-8869
  28. Psahoulia, F. H., K. G. Drosopoulos, L. Doubravska, L. Andera, and A Pintzas. 2007. Quercetin enhances TRAIL-mediated apoptosis in colon cancer cells by inducing the accumulation of death receptors in lipid rafts. Mol. Cancer Ther. 6, 2591-2599
  29. Rieger, L B. Frank M. Weller, and W. Wick. 2007. Mechanisms of resistance of human glioma cells to Apo2 ligand/TNF-related apoptosis-inducing ligand. Cell Physiol. Biochem. 20, 23-34
  30. Russo, M., P. Nigro, R Rosiello, R D' Arienzo, and G. L. Russo. 2007. Quercetin enhances CD95- and TRAIL-induced apoptosis in leukemia cell lines. Leukemia 21, 1130-1133
  31. Siegelin, M. D., D. E. Reuss, A HabeL A Rami, and A von Deimling. 2009. Quercetin promotes degradation of survivin and thereby enhances death-receptor-mediated apoptosis in glioma cells. Neuro. Oncol. 11, 122-131
  32. Siegmund, D., P. Hadwiger, K. Pfizenmaier, H P. Vornlocher, and H Wajant. 2002. Selective inhibition of FUCE-like inhibitory protein expression with small interfering RNA oligonucleotides is sufficient to sensitize tumor cells for TRAIL-induced apoptosis. Mol. Med. 8, 725-732
  33. Shi, RX., C. N. Ong, and H M. Shen. 2004. Luteolin sensitizes tumor necrosis factor-alpha apoptosis in human tumor cells. Oncogene 23, 7712-7721
  34. Syed, V., K. Mukherjee, S. Godoy-Tundidor, and S. M. Ho. 2007. Progesterone induces apoptosis in TRAIL-resistant ovarian cancer cells by circumventing C-FUPL overexpression. J. Cell Biochem. 102, 442-452
  35. Thakkar, H, X. Chen, F. Tyan, S. Gim, H Robinson, C. Lee, S. K. Pandey, C. Nwokorie, N. Onwudiwe, and R K. Srivastava. 2001. Pro-survival function of Akt/ protein kinase B in prostate cancer cells. Relationship with TRAIL resistance. J. Biol. Chem. 276, 38361-38369
  36. Tian, X., G. Chen, H Xing, D. Weng, Y. Guo, and D. Ma. 2007. The relationship between the down-regulation of DNA-PKcs or Ku70 and the chemosensitization in human cervical carcinoma cell line HeLa. Oncol. Rep.18, 927-932
  37. Tomasetti, M., M. R Rippo, R Alleva, S. Moretti, L. Andera, J. Neuzil, and A Procopio. 2004. a-Tocopheryl succinate and TRAIL selectively synergise in induction of apoptosis in human malignant mesothelioma cells. Br. J. Cancer 90, 1644-1653
  38. Uchida, M., M. Iwase, S. Takaoka, S. Yoshiba, G. Kondo, H Watanabe, M. Ohashi, M. Nagumo, and S. Shintani. 2007. Enhanced susceptibility to tumor necrosis factor-related apoptosis-inducing ligand-mediated apoptosis in oral squamous cell carcinoma cells treated with phosphatidylinositol 3-kinase inhibitors. Int. J. Oneal. 30, 1163-1171
  39. Ullenhag, G. L A. Mukherjee, N. F. Watson, A. HAl-Attar, J. H Scholefield, and 1. G. Durrant. 2007. Overexpression of FLIPL is an independent marker of poor prognosis in colorectal cancer patients. Clin. Cancer Res. 13, 5070-5075
  40. Weterings, E. and D. J. Chen. 2007. DNA-dependent protein kinase in nonhomologous end joining: a lock with multiple keys? J. Cell Biol. 179, 183-186
  41. Yagita, H, K, Takeda, Y. M., Hayakawa, J. Smyth, and K Okumura. 2004. TRAIL and its receptors as targets for cancer therapy. Cancer Sci. 95, 777-783
  42. Zhong, X. and A. R. Safa. 2007. Phosphorylation of RNA helicase A by DNA-dependent protein kinase is indispensable for expression of the MDR1 gene product P-glycoprotein in multidrug-resistant human leukemia cells. Biochemistry 46, 5766-5775