Accelerating Effects of Quercetin on the $TNF-{\alpha}-Induced$ Apoptosis in MC3T3-E1 Osteoblastic Cells

  • Choi, Yong-Sung (Laboratory of Cell Biology in Department of Orthodontics and Institute of Oral Biosciences, School of Dentistry) ;
  • Chung, Song-Woo (Laboratory of Cell Biology in Department of Orthodontics and Institute of Oral Biosciences, School of Dentistry) ;
  • Jeon, Young-Mi (Laboratory of Cell Biology in Department of Orthodontics and Institute of Oral Biosciences, School of Dentistry) ;
  • Kim, Jong-Ghee (Laboratory of Cell Biology in Department of Orthodontics and Institute of Oral Biosciences, School of Dentistry) ;
  • Lee, Jeong-Chae (Laboratory of Cell Biology in Department of Orthodontics and Institute of Oral Biosciences, School of Dentistry,Research Center of Bioactive Materials, Chonbuk National University)
  • Published : 2005.09.01

Abstract

Bioflavone quercetin is believed to play an important role preventing bone loss by affecting osteoclastogenesis and regulating many systemic and local factors including hormones and cytokines. This study examined how quercetin acts on tumor necrosis factor-alpha ($TNF-{\alpha}$)-mediated apoptosis in MC3T3-E1 osteoblastic cells. Apoptosis assays revealed the dose-dependent acceleration of quercetin on $TNF-{\alpha}-induced$ apoptosis in MC3T3-E1 cells, which was demonstrated by the increased number of positively stained cells in the trypan blue staining and TUNEL assay, and the migration of many cells to the $sub-G_0/G_1$ phase in flow cytometric analysis. In particular, quercetin treatment alone increased the expression of p53 and p21 proteins in the cells. Consequently, this study showed that quercetin accelerates the $TNF-{\alpha}-induced$ apoptosis in MC3T3-E1 osteoblastic cells.

Keywords

References

  1. Ackland, M.L., van de Waarsenburg, S., and Jones, R., Synergistic anti proliferative action of the flavonols quercetin and kaempferol in cultured human cancer cell lines. In Vivo 19, 69-76 (2005)
  2. Adams, P.D. and Kaelin, W.G. Jr., Negative control elements of the cell cycle in human tumors. Curr. Opin. Cell Biol. 10, 791-797 (1998) https://doi.org/10.1016/S0955-0674(98)80123-3
  3. Allen, R.T., Hunter, W.J. 3rd., and Agrawal, D.K., Morphological and biochemical characterization and analysis of apoptosis. J. Pharmacol. Toxicol. Methods 37, 215-228 (1997) https://doi.org/10.1016/S1056-8719(97)00033-6
  4. Bode, A.M. and Dong, Z., Signal transduction pathways: Targets for green and black tea polyphenols. J. Biochem. Mol. Biol. 36, 66-77 (2003) https://doi.org/10.5483/BMBRep.2003.36.1.066
  5. Bradford, M.M., A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248-254 (1976) https://doi.org/10.1016/0003-2697(76)90527-3
  6. Choi, E.M. and Koo, S.J., Effects of soybean ethanol extract on the cell survival and oxidative stress in osteoblastic cells. Phytother. Res. 17, 627-632 (2003) https://doi.org/10.1002/ptr.1213
  7. Chua, C.C., Chua, B.H., Chen, Z., Landy, C., and Hamdy, R.C., TGF-betal inhibits multiple caspases induced by TNF-${\alpha}$lpha in murine osteoblastic MC3T3-E1 cells. Biochim. Biophys. Acta 1593, 1-8 (2002) https://doi.org/10.1016/S0167-4889(02)00257-4
  8. Franchimont, N., Rydziel, S., Delany, A.M., and Canalis, E., Interleukin-6 and its soluble receptor cause a marked induction of collagenase 3 expression in rat osteoblast cultures. J. Biol. Chem. 272, 12144-12150 (1997) https://doi.org/10.1074/jbc.272.18.12144
  9. Gallagher, J.C., Satpathy, R., Rafferty, K., and Haynatzka, V., The effect of soy protein isolate on bone metabolism. Menopause 11, 290-298 (2004) https://doi.org/10.1097/01.GME.0000097845.95550.71
  10. Gamet-Payrastre, L., Manenti, S., Gratacap, M.P., Tulliez, J., Chap, H., and Payrastre, B., Flavonoids and the inhibition of PKC and PI3-kinase. Gen. Pharmacol. 32, 279-286 (1999) https://doi.org/10.1016/S0306-3623(98)00220-1
  11. Glantschnig, H., Fisher, J.E., Wesolowski, G., Rodan, G.A., and Reszka, A. A. , M-CSF, TNFalpha and RANK ligand promote osteoclast survival by signaling through mTOR/S6 kinase. Cell Death Differ. 10, 1165-1177 (2003) https://doi.org/10.1038/sj.cdd.4401285
  12. Hongo, T., Mizuno, Y, Haraguchi, S., and Yoshida, T.O., A new anticancer drug sensitivity test using the microplate culture and surviving tumor cell staining method. Gan To Kagaku Ryoho 13, 247-254 (1986)
  13. Ishikawa, Y. and Kitamura, M., Anti-apoptotic effect of quercetin: intervention in the JNK- and ERK-mediated apoptotic pathways. Kidney Int. 58, 1078-1087 (2000) https://doi.org/10.1046/j.1523-1755.2000.00265.x
  14. Jeon, Y.M., Kim, B.T., Son, Y.O., Kook, S.H., Lee, K.S., Kim, S.S., Lim, J.Y., Kim, J.G., and Lee, J.C., Effects of quercetin on TNF-${\alpha}$-induced cytokine secretion and nitric oxide production in MC3T3-E1 osteoblastic cells. Nat. Prod. Sci. 11, 103-108 (2005)
  15. Jilka, R.L., Cytokines, bone remodeling, and estrogen deficiency: a 1998 update. Bone 23, 75-81 (1998) https://doi.org/10.1016/S8756-3282(98)00077-5
  16. Kitajima, I., Nakajima, T., Imamura, T., Takasaki, I., Kawahara, K., Okano, T., Tokioka, T., Soejima, Y., Abeyarna, K., and Maruyama, I., Induction of apoptosis in murine clonal osteoblasts expressed by human T-cell leukemia virus type I tax by NF-kappa Band TNF-${\alpha}$lpha. J. Bone Miner. Res. 11, 200-210 (1996)
  17. Lee, J.C., Kim, J., Park, J.K., Chung, G.H., and Jang, Y.S., The antioxidant, rather than prooxidant, activities of quercetin on normal cells: quercetin protects mouse thymocytes from glucose oxidase-mediated apoptosis. Exp. Cell Res. 291, 386-397 (2003) https://doi.org/10.1016/S0014-4827(03)00410-5
  18. Lowe, S.W., Ruley, H.E., Jacks, T., and Housman, D.E., p53-dependent apoptosis modulates the cytotoxicity of anticancer agents. Cell 74, 957-967 (1993) https://doi.org/10.1016/0092-8674(93)90719-7
  19. Miyashita, T., and Reed, J.C., Tumor suppressor p53 is a direct transcriptional activator of the human bax gene. Cell 80, 293-299 (1995) https://doi.org/10.1016/0092-8674(95)90412-3
  20. Monasterio, A., Urdaci, M.C., Pinchuk, I.V., Lopez-Moratalla, N., and Martinez-Irujo, J.J., Flavonoids induce apoptosis in human leukemia U937 cells through caspase- and caspase-calpain-dependent pathways. Nutr. Cancer 50, 90-100 (2004) https://doi.org/10.1207/s15327914nc5001_12
  21. Muller, M., Wilder, S., Bannasch, D., Israeli, D., Lehlbach, K., Li-Weber, M., Friedman, S.L., Galle, P.R., Stremmel, W., Oren, M., and Krammer, P.H., p53 activates the CD95 (APO-1/Fas) gene in response to DNA damage by anticancer drugs. J. Exp. Med. 188, 2033-2045 (1998) https://doi.org/10.1084/jem.188.11.2033
  22. Son, Y.O., Lee, K.Y., Kook, S.H., Lee, J.C., Kim, J.G., Jeon, Y.M., and Jang, Y.S., Selective effects of quercetin on the cell growth and antioxidant defense system in normal versus transformed mouse hepatic cell lines. Eur. J. Pharmacol. 502, 195-204 (2004) https://doi.org/10.1016/j.ejphar.2004.09.012
  23. Suh, K.S., Koh, G., Park, C.Y., Woo, J.T., Kim, S.W., Kim, J.W., Park, I.K., and Kim, Y.S., Soybean isoflavones inhibit tumor necrosis factor-a-induced apoptosis and the production of interleukin-6 and prostaglandin E2 in osteoblastic cells. Phytochemistry 63, 209-215 (2003) https://doi.org/10.1016/S0031-9422(03)00101-8
  24. Wattel, A., Kamel, S., Mentaverri, R., Lorget, F., Prouillet, C., Petit, J.P., Fardelonne, P., and Brazier, M., Potent inhibitory effect of naturally occurring flavonoids quercetin and kaempferol on in vitro osteoclastic bone resorption. Biochem. Pharmacol. 65, 35-42 (2003) https://doi.org/10.1016/S0006-2952(02)01445-4
  25. Woo, J.T., Nakagawa, H., Notoya, M., Yonezawa, T., Udagawa, N., Lee, I.S., Ohnishi, M., Hagiwara, H., and Nagai, K., Quercetin suppresses bone resorption by inhibiting the differentiation and activation of osteoclasts. Biol. Pharm. Bull. 27, 504-509 (2004) https://doi.org/10.1248/bpb.27.504
  26. Yang, E.B., Zhang, K., Cheng, L.Y., and Mack, P., Butein, a specific protein tyrosine kinase inhibitor. Biochem. Biophys. Res. Commun. 245, 435-438 (1998) https://doi.org/10.1006/bbrc.1998.8452