Induction of Apoptosis in Human Leukemic Cell Lines by Diallyl Disulfide via Modulation of EGFR/ERK/PKM2 Signaling Pathways

  • Luo, Nian (Laboratory of Stem Cells and Tissue Engineering, Histology and Embryology, Chongqing Medical University) ;
  • Zhao, Lv-Cui (Laboratory of Stem Cells and Tissue Engineering, Histology and Embryology, Chongqing Medical University) ;
  • Shi, Qing-Qiang (Laboratory of Stem Cells and Tissue Engineering, Histology and Embryology, Chongqing Medical University) ;
  • Feng, Zi-Qiang (Laboratory of Stem Cells and Tissue Engineering, Histology and Embryology, Chongqing Medical University) ;
  • Chen, Di-Long (Laboratory of Stem Cells and Tissue Engineering, Histology and Embryology, Chongqing Medical University) ;
  • Li, Jing (Laboratory of Stem Cells and Tissue Engineering, Histology and Embryology, Chongqing Medical University)
  • Published : 2015.04.29


Background: Diallyl disulfide (DADS) may exert potent anticancer action both in vitro and in vivo. Although its effects on cancer are significant, the underlying mechanisms remain unknown. In this study, we sought to elucidate possible links between DADS and pyruvate kinase (PKM2). Materials and Methods: $KG1{\alpha}$, a leukemia cell line highly expressing PKM2 was used with a cell counting kit (CCK)-8 and flow cytometry (FCM) to investigate the effects of DADS. Relationships between PKM2 and DADS associated with phosphorylation of EGFR, ERK1/2 and MEK, were assessed by western blot analysis. Results: In $KG1{\alpha}$ cells highly expressing PKM2, we found that DADS could affect proliferation, apoptosis and EGFR/ERK/PKM2 signaling pathways, abrogating EGF-induced nuclear accumulation of PKM2. Conclusions: These results suggested that DADS suppressed the proliferation of $KG1{\alpha}$ cells, providing evidence that its proapoptotic effects are mediated through the inhibition of EGFR/ERK/PKM2 signaling pathways.


Supported by : National Natural Science Foundation of China


  1. Aggarwal BB, Shishodia S (2006). Molecular targets of dietary agents for prevention and therapy of cancer. Biochem Pharmacol, 71, 1397-421.
  2. Antony ML, Singh SV (2011). Molecular mechanisms and targets of cancer chemoprevention by garlic-derived bioactive compound diallyl trisulfide. Indian J Exp Biol, 49, 805-16.
  3. Arunkumar AV, Gunadharini N, Krishnamoorthy G, Arunakaran MR (2007). Induction of apoptosis and histone hyperacetylation by diallyl disulfide in prostate cancer cell line PC-3. Cancer Lett, 251, 59-67.
  4. Bautista DM, Movahed P, Hinman A, et al (2005). Pungent products from garlic activate the sensory ion channel TRPA1. Proc Natl Acad Sci U S A, 102, 12248-52.
  5. Block G, Jensen CD, Norkus EP, et al (2007). Usage patterns, health, and nutritional status of long-term multiple dietary supplement users: a cross-sectional study. Nutr J, 6, 30.
  6. Bose CG J, Zimniak L, Srivastava SK, Singh SP, Zimniak P, Singh, S.V2002 (2002). Critical role of allyl groups and disulfide chain in induction of Pi class glutathione transferase in mouse tissues in vivo by diallyl disulfide, a naturally occurring chemopreventive agent in garlic. Carcinogenesis, 23, 1661-5.
  7. Bottone FG Jr, Baek SJ, Nixon JB, Eling TE (2002). Diallyl disulfide (DADS) induces the antitumorigenic NSAIDactivated gene (NAG-1) by a p53-dependent mechanism in human colorectal HCT 116 cells. Nutr J, 132, 773-8.
  8. Fan JM, Liu ZH, Li J, et al (2013). Effect of ginseng polysaccharide-induced wnt/beta-catenin signal transduction pathway on apoptosis of human nasopharyngeal cancer cells CNE-2. Zhongguo Zhong Yao Za Zhi, 38, 3332-7 (in Chinese).
  9. Filomeni GA, K, Rotilio G, Ciriolo, MR (2003). Reactive oxygen species-dependent c-Jun NH2-terminal kinase/c-Jun signaling cascade mediates neuroblastoma cell death induced by diallyl disulfide. Cancer Res, 63, 5940-9.
  10. Frezza C, Pollard PJ, Gottlieb E (2011). Inborn and acquired metabolic defects in cancer. J Mol Med, 89, 213-20.
  11. Frezza CaG, E (2009). Mitochondria in cancer: not just innocent bystanders. Semin Cancer Biol, 19, 4-11.
  12. Gao X, Wang H, Yang JJ, et al (2012). Pyruvate kinase M2 regulates gene transcription by acting as a protein kinase. Mol Cell, 45, 598-609.
  13. Gayathri R, Gunadharini DN, Arunkumar A, et al (2009). Effects of diallyl disulfide (DADS) on expression of apoptosis associated proteins in androgen independent human prostate cancer cells (PC-3). Mol Cell Biochem, 320, 197-203.
  14. Hoshino A, Hirst JA, Fujii H (2007). Regulation of cell proliferation by interleukin-3-induced nuclear translocation of pyruvate kinase. J Biol Chem, 282, 17706-11.
  15. Howard EW, Ling MT, Chua CW, et al (2007). Garlic-derived S-allylmercaptocysteine is a novel in vivo antimetastatic agent for androgen-independent prostate cancer. Clin Cancer Res, 13, 1847-56.
  16. Kwon KBY, SJ, Ryu DG, Yang JY, et al (2002). Induction of apoptosis by diallyl disulfide through activation of caspase-3 in human leukemia HL-60 cells. Biochem Pharmacol, 63, 41-7.
  17. Lee J, Kim HK, Han YM, et al (2008). Pyruvate kinase isozyme type M2 (PKM2) interacts and cooperates with Oct-4 in regulating transcription. Int J Biochem Cell Biol, 40, 1043-54.
  18. Lee JE, Lee RA, Kim KH, et al (2011). Induction of apoptosis with diallyl disulfide in AGS gastric cancer cell line. J Korean Surg Soc, 81, 85-95.
  19. Luo W, Hu H, Chang R, et al (2011). Pyruvate kinase M2 is a PHD3-stimulated coactivator for hypoxia-inducible factor 1. Cell, 145, 732-44.
  20. Mazurek S, Boschek CB, Hugo F, et al (2005). Pyruvate kinase type M2 and its role in tumor growth and spreading. Semin Cancer Biol, 15, 300-8.
  21. Park HSK GY, Choi IW, Kim ND, et al (2011). Inhibition of matrix metalloproteinase activities and tightening of tight junctions by diallyl disulfide in AGS human gastric carcinoma cells. Food Sci, 76, 105-111.
  22. Park HYK ND, Kim GY, Hwang HJ, et al (2012). Inhibitory effects of diallyl disulfide on the production of inflammatory mediators and cytokines in lipopolysaccharide-activated BV2 microglia. Toxicol Appl Pharmacol, 262, 177-184.
  23. Ruvolo PP, Deng X, May WS (2001). Phosphorylation of Bcl2 and regulation of apoptosis. Leukemia, 15, 515-22.
  24. Shin DY, Kim GY, Kim JI, et al (2010). Anti-invasive activity of diallyl disulfide through tightening of tight junctions and inhibition of matrix metalloproteinase activities in LNCaP prostate cancer cells. Toxicol In Vitro, 24, 1569-76.
  25. Stetak A, Veress R, Ovadi J, et al (2007). Nuclear translocation of the tumor marker pyruvate kinase M2 induces programmed cell death. Cancer Res, 67, 1602-8.
  26. Sundaram SGM, J.A (1996). Diallyl disulfide suppresses the growth of human colon tumor cell xenografts in athymic nude mice. Nutr J, 126, 1355-61.
  27. Tan HL H, He J, Yi L, et al (2008). Inhibition of ERK and activation of p38 are involved in diallyl disulfide induced apoptosis of leukemia HL-60 cells. Arch Pharm Res, 31, 786-93.
  28. Tsubura AL YC, Kuwata M, Uehara N, Yoshizawa K (2011). Anticancer effects of garlic and garlic-derived compounds for breast cancer control. Anticancer Agents Med, 11, 249-53.
  29. Warburg O (1956). On the origin of cancer cells. Cancer Biol, 123, 309-14.
  30. Wen JZ Y, Chen X, Shen L, Li GC, Xu M (2004). Enhancement of diallyl disulfide-induced apoptosis by inhibitors of MAPKs in human HepG2 hepatoma cells. Biochem Pharmacol, 68, 323-31.
  31. Xiao DC S, Johnson DE, Vogel VG, et al(2004). Diallyl trisulfide-induced apoptosis in human prostate cancer cells involves c-Jun N-terminal kinase and extracellular-signal regulated kinase-mediated phosphorylation of Bcl-2. Oncogene, 23, 5594-606.
  32. Yang W, Xia Y, Cao Y, et al (2012a). EGFR-induced and PKCepsilon monoubiquitylation-dependent NF-kappaB activation upregulates PKM2 expression and promotes tumorigenesis. Mol Cell, 48, 771-84.
  33. Yang W, Xia Y, Hawke D, et al (2012b). PKM2 phosphorylates histone H3 and promotes gene transcription and tumorigenesis. Cell, 150, 685-96.
  34. Yang W, Xia Y, Ji H, et al (2011). Nuclear PKM2 regulates beta-catenin transactivation upon EGFR activation. Nature, 480, 118-22.
  35. Yang W, Zheng Y, Xia Y, et al (2012c). ERK1/2-dependent phosphorylation and nuclear translocation of PKM2 promotes the Warburg effect. Nat Cell Biol, 14, 1295-304.
  36. Zou H, Henzel WJ, Liu X, et al (1997). Apaf-1, a human protein homologous to C. elegans CED-4, participates in cytochrome c-dependent activation of caspase-3. Cell, 90, 405-13.

Cited by

  1. Subcellular localization of DJ-1 in human HL-60 leukemia cells in response to diallyl disulfide treatment vol.14, pp.5, 2016,
  2. Diallyl disulphide inhibits apolipoprotein(a) expression in HepG2 cells through the MEK1-ERK1/2-ELK-1 pathway vol.16, pp.1, 2017,