Curcumin Inhibits MHCC97H Liver Cancer Cells by Activating ROS/TLR-4/Caspase Signaling Pathway

  • Li, Pei-Min ;
  • Li, Yu-Liang ;
  • Liu, Bin ;
  • Wang, Wu-Jie ;
  • Wang, Yong-Zheng ;
  • Li, Zheng
  • Published : 2014.03.01


Curcumin can inhibit proliferation of liver cancer cells by inducing apoptosis, but the specific signaling pathways involved are not completely clear. Here, we report that curcumin inhibited proliferation of MHCC97H liver cancer cells by induction of apoptosis in a concentration dependent manner via stimulating intracellular reactive oxygen species (ROS) generation. Also, we showed that increased intracellular ROS formation activated the TLR-4/MyD-88 signaling pathway, resulting in activation of caspase-8 and caspase-3, which eventually led to apoptosis in MHCC97H cells. These results showed that as an prooxidant, curcumin exerts anti-cancer effects by inducing apoptosis via the TLR-4/MyD-88 signaling pathway.


Curcumin;liver cancer;reactive oxygen species;toll-like receptor 4;apoptosis


  1. Tan ML, Ooi JP, Ismail N, Moad AI, Muhammad T.S. (2009). Programmed cell death pathways and current antitumor targets. Pharm Res, 26, 1547-60.
  2. Yoshino M, Haneda M, Naruse M, et al (2004). Prooxidant activity of curcumin: copper-dependent formation of 8-hydroxy-2'-deoxyguanosine in DNA and induction of apoptotic cell death. Toxicol In Vitro, 18, 783-9.
  3. Yu T, Sheu SS, Robotham JL, Yoon Y, (2008). Mitochondrial fission mediates high glucose-induced cell death through elevated production of reactive oxygen species. Cardiovasc Res, 79, 341-51.
  4. Zanotto-Filho A, Braganhol E, Edelweiss MI, et al (2012). The curry spice curcumin selectively inhibits cancer cells growth in vitro and in preclinical model of glioblastoma. J Nutr Biochem, 23, 591-601.
  5. Shakibaei M, Mobasheri A, Lueders C, et al (2013). Curcumin enhances the effect of chemotherapy against colorectal cancer cells by inhibition of NF-kappaB and Src protein kinase signaling pathways. PLoS One, 8, e57218.
  6. Simiantonaki N, Kurzik-Dumke U, Karyofylli G, et al (2007). Reduced expression of TLR4 is associated with the metastatic status of human colorectal cancer. Int J Mol Med, 20, 21-9.
  7. Torimura T, Iwamoto H, Nakamura T, et al (2013). Metronomic Chemotherapy: Possible Clinical Application in Advanced Hepatocellular Carcinoma. Transl Oncol, 6, 511-9.
  8. Venook AP, Papandreou C, Furuse J, de Guevara LL (2010). The incidence and epidemiology of hepatocellular carcinoma: a global and regional perspective. Oncologist, 15 Suppl 4, 5-13.
  9. Woo JH, Kim YH, Choi YJ, et al (2003). Molecular mechanisms of curcumin-induced cytotoxicity: induction of apoptosis through generation of reactive oxygen species, downregulation of Bcl-XL and IAP, the release of cytochrome c and inhibition of Akt. Carcinogenesis, 24, 1199-208.
  10. Wu SH, Hang LW, Yang JS, et al (2010). Curcumin Induces Apoptosis in Human Non-small Cell Lung Cancer NCI-H460 Cells through ER Stress and Caspase Cascadeand Mitochondria-dependent Pathways. Anticancer Res, 30, 2125-33.
  11. Yang CL, Ma YG, Xue YX, et al (2012). Curcumin induces small cell lung cancer NCI-H446 cell apoptosis via the reactive oxygen species-mediated mitochondrial pathway and not the cell death receptor pathway. DNA Cell Biol, 31, 139-50.
  12. Nakahira K, Kim HP, Geng XH, et al (2006). Carbon monoxide differentially inhibits TLR signaling pathways by regulating ROS-induced trafficking of TLRs to lipid rafts. J Exp Med, 203, 2377-89.
  13. Matsuzawa A, Saegusa K, Noguchi T, et al (2005). ROSdependent activation of the TRAF6-ASK1-p38 pathway is selectively required for TLR4-mediated innate immunity. Nature Immunology, 6, 587-92.
  14. Mukhopadhyay A, Banerjee S, Stafford LJ, et al (2002). Curcumin-induced suppression of cell proliferation correlates with down-regulation of cyclin D1 expression and CDK4-mediated retinoblastoma protein phosphorylation. Oncogene, 21, 8852-61.
  15. Mishra B, Priyadarsini KI, Bhide MK, Kadam RM, Mohan H (2004). Reactions of superoxide radicals with curcumin: probable mechanisms by optical spectroscopy and EPR. Free Radic Res, 38, 355-62.
  16. Morin D, Barthelemy S, Zini R, Labidalle S, Tillement JP (2001). Curcumin induces the mitochondrial permeability transition pore mediated by membrane protein thiol oxidation. Febs Letters, 495, 131-6.
  17. Pan W, Yang H, Cao C, et al (2008). AMPK mediates curcumininduced cell death in CaOV3 ovarian cancer cells. Oncol Rep, 20, 1553-9.
  18. Parkin DM, Bray F, Ferlay J, Pisani P (2005). Global cancer statistics, 2002. CA Cancer J Clin, 55, 74-108.
  19. Powers KA, Szaszi K, Khadaroo RG, et al (2006). Oxidative stress generated by hemorrhagic shock recruits Toll-like receptor 4 to the plasma membrane in macrophages. J Exp Med, 203, 1951-61.
  20. Qian YG, Deng JF, Geng L, et al (2008). TLR4 signaling induces B7-H1 expression through MAPK pathways in bladder cancer cells. Cancer Investigation, 26, 816-21.
  21. Scott DW, Loo G, (2004). Curcumin-induced GADD153 gene up-regulation in human colon cancer cells. Carcinogenesis, 25, 2155-64.
  22. Kelly MG, Alvero AB, Chen R, et al (2006). TLR-4 signaling promotes tumor growth and paclitaxel chemoresistance in ovarian cancer. Cancer Res, 66, 3859-68.
  23. Kang SK, Cha SH, Jeon HG (2006). Curcumin-induced histone hypoacetylation enhances caspase-3-dependent glioma cell death and neurogenesis of neural progenitor cells. Stem Cells Dev, 5, 165-174.
  24. Kawai T, Akira S (2007). TLR signaling. Semin Immunol, 19, 24-32.
  25. Karunagaran D, Rashmi R, Kumar TR (2005). Induction of apoptosis by curcumin and its implications for cancer therapy. Curr Cancer Drug Targets, 5, 117-29.
  26. Khar A, Ali AM, Pardhasaradhi BVV, Begum Z, Anjum R (1999). Antitumor activity of curcumin is mediated through the induction of apoptosis in AK-5 tumor cells. Febs Letters, 445, 165-8.
  27. Liu D, Chen Z (2013). The effect of curcumin on breast cancer cells. J Breast Cancer, 16, 133-7.
  28. Liu ZW, Zhu HT, Chen KL, et al (2013). Protein kinase RNAlike endoplasmic reticulum kinase (PERK) signaling pathway plays a major role in reactive oxygen species (ROS)- mediated endoplasmic reticulum stress- induced apoptosis in diabetic cardiomyopathy. Cardiovasc Diabetol, 12, 158.
  29. Lopez-Lazaro M (2008). Anticancer and carcinogenic properties of curcumin: Considerations for its clinical development as a cancer chemopreventive and chemotherapeutic agent. Mol Nutr Food Res, 52, S103-27.
  30. Lv J, Jia R, Yang D, Zhu J, Ding G (2009). Candesartan attenuates Angiotensin II-induced mesangial cell apoptosis via TLR4/MyD88 pathway. Biochem Biophys Res Commun, 380, 81-6
  31. Martin-Cordero C, Leon-Gonzalez AJ, Calderon-Montano JM, Burgos-Moron E, Lopez-Lazaro M (2012). Pro-oxidant natural products as anticancer agents. Current Drug Targets, 13, 1006-28.
  32. Hatcher H, Planalp R, Cho J, Torti FM, Torti SV (2008). Curcumin: from ancient medicine to current clinical trials. Cell Mol Life Sci, 65, 1631-52.
  33. Debatin KM (2004). Apoptosis pathways in cancer and cancer therapy. Cancer Immunol Immunother, 53, 153-9.
  34. Gupta S (2003). Molecular signaling in death receptor and mitochondrial pathways of apoptosis (Review). Int J Oncol, 22, 15-20.
  35. Han KJ, Su XQ, Xu LG, et al (2004). Mechanisms of the TRIFinduced interferon-stimulated response element and NFkappa B activation and apoptosis pathways. J Biol Chem, 279, 15652-61.
  36. Hengartner MO (2000). The biochemistry of apoptosis. Nature, 407, 770-6.
  37. Huang B, Zhao J, Li H, et al (2005). Toll-like receptors on tumor cells facilitate evasion of immune surveillance. Cancer Res, 65, 5009-14.
  38. Huang D, Fang F, Xu F (2011). Hyperoxia-induced up-regulation of Toll-like receptors expression in alveolar epithelial cells. Zhongguo Wei Zhong Bing Ji Jiu Yi Xue, 23, 645-9.
  39. Iqbal M, Okazaki Y, Okada S, (2009). Curcumin attenuates oxidative damage in animals treated with a renal carcinogen, ferric nitrilotriacetate (Fe-NTA): implications for cancer prevention. Mol Cell Biochem, 324, 157-64.
  40. Iqbal M, Sharma SD, Okazaki Y, Fujisawa M, Okada S. (2003). Dietary supplementation of curcumin enhances antioxidant and phase II metabolizing enzymes in ddY male mice: possible role in protection against chemical carcinogenesis and toxicity. Pharmacol Toxicol, 92, 33-8.
  41. Janeway CA, Jr.Medzhitov R. (2002). Innate immune recognition. Annu Rev Immunol, 20, 197-216.
  42. Chen K, Zhang S, Ji Y, et al (2013). Baicalein inhibits the invasion and metastatic capabilities of hepatocellular carcinoma cells via down-regulation of the ERK pathway. PLoS One, 8, e72927.
  43. Atsumi T, Tonosaki K, Fujisawa S (2006). Induction of early apoptosis and ROS-generation activity in human gingival fibroblasts (HGF) and human submandibular gland carcinoma (HSG) cells treated with curcumin. Arch Oral Biol, 51, 913-21.
  44. Benchoua A, Couriaud C, Guegan C, et al (2002). Active caspase-8 translocates into the nucleus of apoptotic cells to inactivate poly (ADP-ribose) polymerase-2. J Biol Chem, 277, 34217-22.
  45. Caldwell S, Park SH (2009). The epidemiology of hepatocellular cancer: from the perspectives of public health problem to tumor biology. J Gastroenterol, 44 Suppl 19, 96-101.
  46. Dai XZ, Yin HT, Sun LF, et al (2013). Potential therapeutic efficacy of curcumin in liver cancer. Asian Pac J Cancer Prev, 14, 3855-9.
  47. Dasu MR, Devaraj S, Zhao L, Hwang DH, Jialal I (2008). High glucose induces toll-like receptor expression in human monocytes: mechanism of activation. Diabetes, 57, 3090-8.
  48. Ahn JC, Kang JW, Shin JI, Chung P.S., (2012). Combination treatment with photodynamic therapy and curcumin induces mitochondria-dependent apoptosis in AMC-HN3 cells. Int J Oncol, 41, 2184-90.
  49. Aggarwal BB, Kumar A, Bharti AC (2003). Anticancer potential of curcumin: preclinical and clinical studies. Anticancer Res, 23, 363-98.
  50. Andreani V, Gatti G, Simonella L, Rivero V, Maccioni M (2007). Activation of Toll-like receptor 4 on tumor cells in vitro inhibits subsequent tumor growth in vivo. Cancer Res, 67, 10519-27.

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