Ganoderma Lucidum Polysaccharides Target a Fas/Caspase Dependent Pathway to Induce Apoptosis in Human Colon Cancer Cells

  • Liang, Zengenni (Hunan Provincial Key Laboratory of Crop Germplasm Innovation and Utilization, Hunan Agricultural University) ;
  • Guo, Yu-Tong (Hunan Provincial Key Laboratory of Crop Germplasm Innovation and Utilization, Hunan Agricultural University) ;
  • Yi, You-Jin (Hunan Provincial Key Laboratory of Crop Germplasm Innovation and Utilization, Hunan Agricultural University) ;
  • Wang, Ren-Cai (Hunan Provincial Key Laboratory of Crop Germplasm Innovation and Utilization, Hunan Agricultural University) ;
  • Hu, Qiu-Long (Hunan Provincial Key Laboratory of Crop Germplasm Innovation and Utilization, Hunan Agricultural University) ;
  • Xiong, Xing-Yao (Hunan Provincial Key Laboratory of Crop Germplasm Innovation and Utilization, Hunan Agricultural University)
  • Published : 2014.05.15


Ganoderma lucidum polysaccharides (GLP) extracted from Ganoderma lucidum have been shown to induce cell death in some kinds of cancer cells. This study investigated the cytotoxic and apoptotic effect of GLP on HCT-116 human colon cancer cells and the molecular mechanisms involved. Cell proliferation, cell migration, lactate dehydrogenase (LDH) levels and intracellular free calcium levels ($[Ca^{2+}]i$) were determined by MTT, wound-healing, LDH release and fluorescence assays, respectively. Cell apoptosis was observed by scanning and transmission electron microscopy. For the mechanism studies, caspase-8 activation, and Fas and caspase-3 expression were evaluated. Treatment of HCT-116 cells with various concentrations of GLP (0.625-5 mg/mL) resulted in a significant decrease in cell viability (P< 0.01). This study showed that the antitumor activity of GLP was related to cell migration inhibition, cell morphology changes, intracellular $Ca^{2+}$ elevation and LDH release. Also, increase in the levels of caspase-8 activity was involved in GLP-induced apoptosis. Western blotting indicated that Fas and caspase-3 protein expression was up-regulated after exposure to GLP. This investigation demonstrated for the first time that GLP shows prominent anticancer activities against the HCT-116 human colon cancer cell line through triggering intracellular calcium release and the death receptor pathway.


  1. Chen XP, Chen Y, Li SB, et al (2009). Free radical scavenging of Ganoderma lucidum polysaccharides and its effect on antioxidant enzymes and immunity activities in cervical carcinoma rats. Carbohyd Polym, 77, 389-93.
  2. Chen XP, Wang WX, Li SB, et al (2010). Optimization of ultrasound-assisted extraction of Lingzhi polysaccharides using response surface methodology and its inhibitory effects on cervical cancer cells. Carbohyd Polym, 80, 944-8.
  3. Chen YG, Shen ZJ, Chen XP (2009). Modulatory effect of Ganoderma lucidum polysaccharides on serum antioxidant enzymes activities in ovarian cancer rats. Carbohyd Polym, 78, 258-62.
  4. Chien CM, Cheng JL, Chang WT, et al (2004). Polysaccharide of Ganoderma lucidum alter cell immunophenotypic expression and enhance $CD56^{+}$ NK-cell cytotoxicity in cord blood. Bioorgan Med Chem, 12, 5603-9.
  5. Condeelis J, Pollard JW (2006). Macrophages: Obligate partners for tumor cell migration, invasion, and metastasis. Cell, 124, 263-6.
  6. Dai Q, Yu M, Lang L, Ji Y (2013). Research on structural modification and structure-activity relationship about anti-tumor of polysaccharides from plants. AMM, 411-414, 3232-6.
  7. Giavasis L (2014). Bioactive fungal polysaccharides as potential functional ingredients in food and nutraceuticals. Curr Opin Biotech, 26, 162-73.
  8. Grynkiewicz G, Poenie M, Tsien RY (1985). A new generation of $Ca^{2+}$ indicators with greatly improved fluorescence properties. J Biol Chem, 60, 3440-50.
  9. Guo YT (2012). Inhibitory effect of Ganoderma lucidum polysaccharides on human colon cancer cells proliferation. Hunan Agriculture University, 12-5 (in Chinese).
  10. Hirst RA, Harrison C, Hirota K, Lambert DG (1999). Measurement of [$Ca^{2+}$]i in whole cell suspensions using Fura-2. Methods Mol Biol, 114, 31-9.
  11. Hsu JW, Huang HC, Chen ST, et al (2011). Ganoderma lucidum polysaccharides induce macrophage-like differentiation in human leukemia THP-1 cells via caspase and p53 activation. Evid-Based Compl Alt, 2011, 358-717.
  12. Jiang ZY, Lin C, Liu XC, et al (2003). Effects of Ganoderma lucidum polysaccharides on the immune function in chickens. J Jinan Univ (Natural Science & Medicine Edition), 24, 51-3 (in Chinese).
  13. Li FL, Zhang YM, Zhong ZJ (2011). Antihyperglycemic effect of Ganoderma lucidum polysaccharides on streptozotocin-induced diabetic mice. Int J Mol Sci, 12, 6135-45.
  14. Li WJ, Nie SP, Xie MY, et al (2011). Ganoderma atrum polysaccharide attenuates oxidative stress induced by D-galactose in mouse brain. Life Sci, 88, 713-8.
  15. Liang Z, Yi YJ, Guo YT, et al (2012). Effect of combined Ganoderma lucidum polysacchrides and fluorouracil on proliferation and apoptosis in human colon carcinoma HCT-116 cells. Food Sci, 33, 310-4 (in Chinese).
  16. Lin CY, Chen YH, Lin CY, et al (2010). Ganoderma lucidum polysaccharides attenuate endotoxin-induced intercellular cell adhesion molecule-1 expression in cultured smooth muscle cells and in the neointima in mice. J Agric Food Chem, 58, 9563-71.
  17. Liu L, Cao Y, Chen C, et al (2006). Sorafenib blocks the RAF/MEK/ERK pathway, inhibits tumor angiogenesis, and induces tumor cell apoptosis in hepatocellular carcinoma model PLC/PRF/5. Cancer Res, 66, 11851-8.
  18. Liu W, Wang HY, Pang XB, et al (2010). Characterization and antioxidant activity of two low-molecular-weight polysaccharides purified from the fruiting bodies of Ganoderma lucidum. Int J Biol Macromol, 46, 451-7.
  19. Liu WH, Chang LS (2011). Fas/FasL-dependent and -independent activation of caspase-8 in doxorubicin-treated human breast cancer MCF-7 cells: ADAM10 down-regulation activates Fas/FasL signaling pathway. Int J Biochem Cell Biol, 43, 1708-19.
  20. Luigi S, Calogero C, Fabio F, et al (2009). Minor hepatic resection using heat coagulative necrosis. Am Surg, 75, 1213-9.
  21. Mandal D, Mazumder A, Das P, et al (2005). Fas-, caspase 8-, and caspase 3-dependent signaling regulates the activity of the aminophospholipid translocase and phosphatidylserine externalization in human erythrocytes. J Biol Chem, 280, 39460-7.
  22. Nakagawa T, Shimizu S, Watanabe T, et al (2005). Cyclophilin D-dependent mitochondrial permeability transition regulates some necrotic but not apoptotic cell death. Nature, 434, 652-8.
  23. Ning X, Liu Q, Li C, et al (2014). Inhibitory effects of a polysaccharide extract from the Chaga medicinal mushroom, Inonotus obliquus (higher basidiomycetes), on the proliferation of human neurogliocytoma cells. Int J Med Mushrooms, 16, 29-36.
  24. Nunez R, Sancho-Martinez SM, Novoa JML, Lopez-Hernandez J (2010). Apoptotic volume decrease as a geometric determinant for cell dismantling into apoptotic bodies. Cell Death Differ, 17, 1665-71.
  25. Pillai TG, Nair CKK, Janardhanan KK (2010). Enhancement of repair of radiation induced DNA strand breaks in human cells by Ganoderma mushroom polyssaccharides. Food Chem, 119, 1040-3.
  26. Sayers TJ (2011). Targeting the extrinsic apoptosis signaling pathway for cancer therapy. Cancer Immunol Immunother, 2011, 60, 1173-80.
  27. Shang D, Li Y, Wang C, et al (2011). A novel polysaccharide from se-enriched Ganoderma lucidum induces apoptosis of human breast cancer cells. Oncol Rep, 25, 267-72.
  28. Shi X, Zhao Y, Jiao Y, et al (2013). ROS-dependent mitochondria molecular mechanisms underlying antitumor activity of Pleurotus abalonus acidic polysaccharides in human breast cancer MCF-7 cells. Plos One, 8, e64266.
  29. Sun B, Cai YY, Li YS, et al (2013). The nonstructural protein NP1 of human bocavirus 1 induces cell cycle arrest and apoptosis in Hela cells. Virology, 440, 75-83.
  30. Sun LX, Lin ZB, Duan XS, et al (2011). Ganoderma lucidum polysaccharides antagonize the suppression on lymphocytes induced by culture supernatants of B16F10 melanoma cells. J Pharm Pharmacol, 63, 725-35.
  31. Suresh S, Raaghu D, Karunagaran D (2013). Menadione (Vitamin K3) induces apoptosis of human oral cancer cells and reduces their metastatic potential by modulating the expression of epithelial to mesenchymal transition markers and inhibiting migration. Asian Pac J Cancer Prev, 14, 5461-5.
  32. Wahab SIA, Abdul AB, Alzubairi AS, et al (2009). In vitro ultramorphological assessment of apoptosis induced by Zerumbone on (Hela). J Biomed Biotechnol, 2009, 769568.
  33. Yamaguchi H, Condeelis J (2007). Regulation of the actin cytoskeleton in cancer cell migration and invasion. BBA-Mol Cell Res, 1773, 642-52.
  34. Zhao GH, Kan IQ, Li ZX, Chen ZD (2005). Structural features and immunological activity of a polysaccaride from Dioscorea opposita thunb root. Carbohyd Res, 61, 125-31.
  35. Zhou GQ, Zhao HY, Lu C, et al (2009). Effect of Ganoderma lucidum polysaccharides on intestinal mucosal immune system in H22 liver cancer bearing mice. Chin J Integr Med, 29, 335-9.

Cited by

  1. Research Progress on the Antineoplastic Pharmacological Effects and Mechanisms of Litchi Seeds vol.06, pp.01, 2015,
  2. Effects of Two Traditional Chinese Cooking Oils, Canola and Pork, on pH and Cholic Acid Content of Faeces and Colon Tumorigenesis in Kunming Mice vol.16, pp.15, 2015,
  3. Ganoderma vol.25, pp.5, 2016,
  4. Restoration of the tumor-suppressor function to mutant p53 by Ganoderma lucidum polysaccharides in colorectal cancer cells vol.37, pp.1, 2016,
  5. Antitumor Potential of Substances from the Fungus Ganoderma lucidum vol.52, pp.1, 2018,