Abstract
Aim:
To investigate whether subtoxic concentration of β-sitosterol (SITO) combined with TNF-related apoptosis-inducing ligand (TRAIL) induces apoptosis in TRAIL-resistant MDA-MB-231 breast cancer cells.
Methods:
Cell viability and growth were assessed by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphnyl-2H-tetrazolim bromide assays, chromatin condensation, release of lactate dehydrogenase (LDH), and Annexin V+ cells. The apoptosis-related proteins were detected by Western blotting.
Results:
Treatment with TRAIL in combination with subtoxic concentrations of SITO sensitized MDA-MB-231 breast cancer cells to TRAIL-mediated apoptosis. The synergistic treatment induced chromatin condensation, DNA fragmentation, the release of LDH, and Annexin V+ cells. The indicators of apoptosis are correlated to the induction of caspase activities, which results in the cleavage of poly(ADP-ribose)polymerase. Both the cytotoxic effects and apoptotic characteristics induced by the synergistic treatment were significantly inhibited by a pan-caspase inhibitor z-VAD-fmk, demonstrating the important role of caspases. These results indicate that caspases are crucial regulators of apoptosis induced by the combined treatment of SITO and TRAIL in MDA-MB-231 cells.
Conclusion:
The synergistic treatment of SITO and TRAIL induces apoptosis, which can serve as a potential preventive and therapeutic agent.
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Ashkenazi A, Pai RC, Fong S, Leung S, Lawrence DA, Marsters SA, et al. Safety and antitumor activity or recombinant soluble Apo2 ligand. J Clin Invest 1999; 104: 155–62.
Walczak H, Miller RE, Ariail K, Gliniak B, Griffth TS, Kubin M, et al. Tumoricidal activity of tumor necrosis factor-related apoptosis-inducing ligand in vivo. Nat Med 1999; 5: 157–63.
Pan G, Ni J, Wei YF, Yu G, Gentz R, Dixit VM . An antagonist decoy receptor and a death domain-containing receptor for TRAIL. Science 1997; 277: 815–8.
Pan G, O'Rourke K, Chinnaiyan AM, Gentz R, Ebner R, Ni J, et al. The receptor for the cytotoxic ligand TRAIL. Science 1997; 276: 111–3.
Srivastava RK . TRAIL/Apo-2L: mechanisms and clinical applications in cancer. Neoplasia 2001; 3: 535–46.
Singh TR, Shankar S, Chen X, Asim M, Srivastava RK . Synergistic interactions of chemotherapeutic drugs and tumor necrosis factor-related apoptosis-inducing ligand/Apo-2 ligand on apoptosis and on regression of breast carcinoma in vivo. Cancer Res 2003; 63: 5390–400.
Shao R, Lee DF, Wen Y, Ding Y, Xia W, Ping B, et al. E1A sensitizes cancer cells to TRAIL-induced apoptosis through enhancement of caspase activation. Mol Cancer Res 2005; 3: 219–26.
Butler LM, Liapis V, Bouralexis S, Welldon K, Hay S, Thai LM, et al. The histone deacetylase inhibitor, suberoylanilide hydroxamic acid, overcomes resistance of human breast cancer cells to Apo2L/TRAIL. Int J Cancer 2006; 119: 944–54.
Awad AB, Fink CS . Phytosterols as anticancer dietary components: evidence and mechanism of action. J Nutr 2000; 130: 2127–30.
Vanhanen HT, Miettinen TA . Effects of unsaturated and saturated dietary plant sterols on their serum contents. Clin Chim Acta 1992; 205: 97–107.
Muti P, Awad AB, Schunemann H, Fink CS, Hovey K, Freudenheim JL, et al. A plant food-based diet modifes the serum β-sitosterol concentration in hyperandrogenic postmenopausal women. J Nutr 2003; 133; 4252–55.
Raicht RF, Cohen BI, Fazzini EP, Sarwal AN, Takahashi M . Protective effect of plant sterols against chemically induced colon tumors in rats. Cancer Res 1980; 40: 403–5.
Gupta MB, Nath R, Srivastava N, Shanker K, Kishor K, Bhargava KP . Anti-inflammatory and antipyretic activities of β-sitosterol. Planta Med 1980; 39: 157–63.
Garcia MD, Saenz MT, Gomez MA, Fernandez MA . Topical anti-inflammatory activity of phytosterols isolated from Eryngium foetidum on chronic and acute inflammation models. Phytother Res 1999; 13: 78–80.
Choi S, Kim KW, Choi JS, Han ST, Park YI, Lee SK, et al. Angiogenic activity of β-sitosterol in the ischemia/reperfusion-damaged brain of Mongolian gerbil. Planta Med 2002; 68: 330–5.
Bouic PJ, Lamprecht JH . Plant sterols and sterolins: a review of their immune-modulating properties. Altern Med Rev 1999; 4: 170–7.
Bouic PJ . The role of phytosterols and phytosterolins in immune modulation: a review of the past 10 years. Curr Opin Clin Nutr Metab Care 2001; 4: 471–5.
Clodi K, Wimmer D, Li Y, Goodwin R, Jaeger U, Mann G, et al. Expression of tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) receptors and sensitivity to TRAIL-induced apoptosis in primary B-cell acute lymphoblastic leukemia cells. Br J Haematol 2000; 111: 580–6.
Fulda S, Jeremias I, Debatin KM . Cooperation of betulinic acid and TRAIL to induce apoptosis in tumor cells. Oncogene 2004; 23: 7611–20.
MacFarlane M, Harper N, Snowden RT, Dyer MJ, Barnett GA, Pringle JH, et al. Mechanisms of resistance to TRAIL-induced apoptosis in primary B cell chronic lymphocytic leukemia. Oncogene 2002; 21: 6809–18.
Awad AB, Roy R, Fink CS . β-Sitosterol, a plant sterol, induces apoptosis and activates key caspases in MDA-MB-231 human breast cancer cells. Oncol Rep 2003; 10: 497–500.
Awad AB, Williams H, Fink CS . Phytosterols reduce in vitro metastatic ability of MDA-MB-231 human breast caner cells. Nutr Cancer 2001; 40: 157–64.
Awad AB, Downie A, Fink CS, Kim U . Dietary phytosterol inhibits the growth and metastasis of MDA-MB-231 human breast cancer cells grown in SCID mice. Anticancer Res 2000; 20: 821–4.
Awad AB, Downie AC, Fink CS . Induction of growth and stimulation of apoptosis by β-sitosterol treatment of MDA-MB-231 human breast cancer cells in culture. Int J Mol Med 2000; 5: 541–45.
Sprick MR, Weigand MA, Rieser E, Rauch CT, Juo P, Blenis J, et al. FADD/MORT1 and caspase-8 are recruited to TRAIL receptors 1 and 2 are essential for apoptosis mediated by TRAIL receptor 2. Immunity 2000; 12: 599–609.
Bratton SB, MacFarlane M, Cain K, Cohen GM . Protein complexes activate distinct caspase cascades in death receptor and stress-induced apoptosis. Exp Cell Res 2000; 256: 27–33.
Cohen GM . Caspases: the executioners of apoptosis. Biochem J 1997; 326: 1–16.
Stennicke HR, Salvesen GS . Properties of the caspases. Biochim Biophys Acta 1998; 1387: 17–31.
Earnshaw WC, Martins LM, Kaufmann SH . Mammalian caspases: structure, activation, substrates, and functions during apoptosis. Annu Rev Biochem 1999; 68: 383–424.
Yang LQ, Fang DC, Wang RQ, Yang SM . Effect of NF-κB, survivin, Bc1-2 and caspase3 on apoptosis of gastric cancer cells induced by tumor necrosis factor related apoptosis inducing ligand. World J Gastroenterol 2004; 10: 22–5.
LeBlanc H, Lawrence D, Varfolomeev E, Totpal K, Morlan J, Schow P, et al. Tumor-cell resistance to death receptor-induced apoptosis through mutational inactivation of the proapoptotic Bc1-2 homolog Bax. Nat Med 2002; 8: 274–81.
Kim EH, Kim SU, Shin DY, Choi KS . Roscovitine sensitizes glioma cells to TRAIL-mediated apoptosis by downregulation of survivin and XIAP. Oncogene 2004; 23: 446–56.
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This study was supported by a grant of the Korea Health 21 R&D Project, Ministry of Health & Welfare, Republic of Korea (No A062406).
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Park, C., Moon, Do., Ryu, Ch. et al. β-Sitosterol sensitizes MDA-MB-231 cells to TRAIL-induced apoptosis. Acta Pharmacol Sin 29, 341–348 (2008). https://doi.org/10.1111/j.1745-7254.2008.00761.x
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DOI: https://doi.org/10.1111/j.1745-7254.2008.00761.x
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