Molecules and Cells

  • 제38권2호
  • /
  • Pages.138-144
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  • 2015
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  • 1016-8478(pISSN)
  • /
  • 0219-1032(eISSN)
한국분자세포생물학회 (Korean Society for Molecular and Cellular Biology)
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Raloxifene Induces Autophagy-Dependent Cell Death in Breast Cancer Cells via the Activation of AMP-Activated Protein Kinase

  • Kim, Dong Eun (Institute for Innovative Cancer Research, University of Ulsan College of Medicine, Asan Medical Center) ;
  • Kim, Yunha (Institute for Innovative Cancer Research, University of Ulsan College of Medicine, Asan Medical Center) ;
  • Cho, Dong-Hyung (Graduate School of East-West Medical Science, Kyung Hee University) ;
  • Jeong, Seong-Yun (Institute for Innovative Cancer Research, University of Ulsan College of Medicine, Asan Medical Center) ;
  • Kim, Sung-Bae (Department of Oncology, University of Ulsan College of Medicine, Asan Medical Center) ;
  • Suh, Nayoung (Asan Institute for Life Sciences, University of Ulsan College of Medicine, Asan Medical Center) ;
  • Lee, Jung Shin (Institute for Innovative Cancer Research, University of Ulsan College of Medicine, Asan Medical Center) ;
  • Choi, Eun Kyung (Institute for Innovative Cancer Research, University of Ulsan College of Medicine, Asan Medical Center) ;
  • Koh, Jae-Young (Neural Injury Research Center and Department of Neurology, Asan Medical Center) ;
  • Hwang, Jung Jin (Institute for Innovative Cancer Research, University of Ulsan College of Medicine, Asan Medical Center) ;
  • Kim, Choung-Soo (Institute for Innovative Cancer Research, University of Ulsan College of Medicine, Asan Medical Center)
  • 투고 : 2014.07.09
  • 심사 : 2014.11.10
  • 발행 : 2015.02.28
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초록

Raloxifene is a selective estrogen receptor modulator (SERM) that binds to the estrogen receptor (ER), and exhibits potent anti-tumor and autophagy-inducing effects in breast cancer cells. However, the mechanism of raloxifene-induced cell death and autophagy is not well-established. So, we analyzed mechanism underlying death and autophagy induced by raloxifene in MCF-7 breast cancer cells. Treatment with raloxifene significantly induced death in MCF-7 cells. Raloxifene accumulated GFP-LC3 puncta and increased the level of autophagic marker proteins, such as LC3-II, BECN1, and ATG12-ATG5 conjugates, indicating activated autophagy. Raloxifene also increased autophagic flux indicators, the cleavage of GFP from GFP-LC3 and only red fluorescence-positive puncta in mRFP-GFP-LC3-expressing cells. An autophagy inhibitor, 3-methyladenine (3-MA), suppressed the level of LC3-II and blocked the formation of GFP-LC3 puncta. Moreover, siRNA targeting BECN1 markedly reversed cell death and the level of LC3-II increased by raloxifene. Besides, raloxifene-induced cell death was not related to cleavage of caspases-7, -9, and PARP. These results indicate that raloxifene activates autophagy-dependent cell death but not apoptosis. Interestingly, raloxifene decreased the level of intracellular adenosine triphosphate (ATP) and activated the AMPK/ULK1 pathway. However it was not suppressed the AKT/mTOR pathway. Addition of ATP decreased the phosphorylation of AMPK as well as the accumulation of LC3-II, finally attenuating raloxifene-induced cell death. Our current study demonstrates that raloxifene induces autophagy via the activation of AMPK by sensing decreases in ATP, and that the overactivation of autophagy promotes cell death and thereby mediates the anti-cancer effects of raloxifene in breast cancer cells.

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참고문헌

  1. Alers, S., Loffler, A.S., Wesselborg, S., and Stork, B. (2012). Role of AMPK-mTOR-Ulk1/2 in the regulation of autophagy: cross talk, shortcuts, and feedbacks. Mol. Cell. Biol. 32, 2-11. https://doi.org/10.1128/MCB.06159-11
  2. Balgi, A.D., Fonseca, B.D., Donohue, E., Tsang, T.C., Lajoie, P., Proud, C.G., Nabi, I.R., and Roberge, M. (2009). Screen for chemical modulators of autophagy reveals novel therapeutic inhibitors of mTORC1 signaling. PLoS One 4, e7124. https://doi.org/10.1371/journal.pone.0007124
  3. Bursch, W., Ellinger, A., Kienzl, H., Torok, L., Pandey, S., Sikorska, M., Walker, R., and Hermann, R.S. (1996). Active cell death induced by the anti-estrogens tamoxifen and ICI 164 384 in human mammary carcinoma cells (MCF-7) in culture: the role of autophagy. Carcinogenesis 17, 1595-1607. https://doi.org/10.1093/carcin/17.8.1595
  4. de Medina, P., Payre, B., Boubekeur, N., Bertrand-Michel, J., Terce, F., Silvente-Poirot, S., and Poirot, M. (2009). Ligands of the antiestrogen- binding site induce active cell death and autophagy in human breast cancer cells through the modulation of cholesterol metabolism. Cell Death Differ. 16, 1372-1384. https://doi.org/10.1038/cdd.2009.62
  5. Deli, T., and Csernoch, L. (2008). Extracellular ATP and cancer: an overview with special reference to P2 purinergic receptors. Pathol. Oncol. Res. 14, 219-231. https://doi.org/10.1007/s12253-008-9071-7
  6. Dixon, C.J., Bowler, W.B., Fleetwood, P., Ginty, A.F., Gallagher, J.A., and Carron, J.A. (1997). Extracellular nucleotides stimulate proliferation in MCF-7 breast cancer cells via P2-purinoceptors. Br. J. Cancer 75, 34-39. https://doi.org/10.1038/bjc.1997.6
  7. Dorsey, F.C., Steeves, M.A., Prater, S.M., Schroter, T., and Cleveland, J.L. (2009). Monitoring the autophagy pathway in cancer. Methods Enzymol. 453, 251-271. https://doi.org/10.1016/S0076-6879(08)04012-3
  8. Egan, D., Kim, J., Shaw, R.J., and Guan, K.L. (2011). The autophagy initiating kinase ULK1 is regulated via opposing phosphorylation by AMPK and mTOR. Autophagy 7, 643-644. https://doi.org/10.4161/auto.7.6.15123
  9. Eskelinen, E.L. (2008). New insights into the mechanisms of macroautophagy in mammalian cells. Int. Rev. Cell Mol. Biol. 266, 207-247. https://doi.org/10.1016/S1937-6448(07)66005-5
  10. Fabian, C.J., and Kimler, B.F. (2005). Selective estrogen-receptor modulators for primary prevention of breast cancer. J. Clin. Oncol. 23, 1644-1655. https://doi.org/10.1200/JCO.2005.11.005
  11. Fisher, B., Costantino, J.P., Wickerham, D.L., Redmond, C.K., Kavanah, M., Cronin, W.M., Vogel, V., Robidoux, A., Dimitrov, N., Atkins, J., et al. (1998). Tamoxifen for prevention of breast cancer: report of the national surgical adjuvant breast and bowel project P-1 study. J. Natl. Cancer Inst. 90, 1371-1388. https://doi.org/10.1093/jnci/90.18.1371
  12. Gizzo, S., Saccardi, C., Patrelli, T.S., Berretta, R., Capobianco, G., Di Gangi, S., Vacilotto, A., Bertocco, A., Noventa, M., Ancona, E., et al. (2013). Update on raloxifene: mechanism of action, clinical efficacy, adverse effects, and contraindications. Obstet. Gynecol. Surv. 68, 467-481. https://doi.org/10.1097/OGX.0b013e31828baef9
  13. He, C., and Klionsky, D.J. (2009). Regulation mechanisms and signaling pathways of autophagy. Annu. Rev. Genet. 43, 67-93. https://doi.org/10.1146/annurev-genet-102808-114910
  14. Hippert, M.M., O'Toole, P.S., and Thorburn, A. (2006). Autophagy in cancer: good, bad, or both? Cancer Res. 66, 9349-9351. https://doi.org/10.1158/0008-5472.CAN-06-1597
  15. Hopfner, M., Lemmer, K., Jansen, A., Hanski, C., Riecken, E.O., Gavish, M., Mann, B., Buhr, H., Glassmeier, G., and Scherubl, H. (1998). Expression of functional P2-purinergic receptors in primary cultures of human colorectal carcinoma cells. Biochem. Biophys. Res. Commun. 251, 811-817. https://doi.org/10.1006/bbrc.1998.9555
  16. Hwang, J.J., Kim, H.N., Kim, J., Cho, D.H., Kim, M.J., Kim, Y.S., Kim, Y., Park, S.J., and Koh, J.Y. (2010). Zinc(II) ion mediates tamoxifen-induced autophagy and cell death in MCF-7 breast cancer cell line. Biometals 23, 997-1013. https://doi.org/10.1007/s10534-010-9346-9
  17. Jung, C.H., Ro, S.H., Cao, J., Otto, N.M., and Kim, D.H. (2010). mTOR regulation of autophagy. FEBS Lett. 584, 1287-1295. https://doi.org/10.1016/j.febslet.2010.01.017
  18. Kanzawa, T., Zhang, L., Xiao, L., Germano, I.M., Kondo, Y., and Kondo, S. (2005). Arsenic trioxide induces autophagic cell death in malignant glioma cells by upregulation of mitochondrial cell death protein BNIP3. Oncogene 24, 980-991. https://doi.org/10.1038/sj.onc.1208095
  19. Khan, J.A., Forouhar, F., Tao, X., and Tong, L. (2007). Nicotinamide adenine dinucleotide metabolism as an attractive target for drug discovery. Expert Opin. Ther. Targets 11, 695-705. https://doi.org/10.1517/14728222.11.5.695
  20. Kim, J., Kundu, M., Viollet, B., and Guan, K.L. (2011). AMPK and mTOR regulate autophagy through direct phosphorylation of Ulk1. Nat. Cell Biol. 13, 132-141. https://doi.org/10.1038/ncb2152
  21. Lee, J.W., Park, S., Takahashi, Y., and Wang, H.G. (2010). The association of AMPK with ULK1 regulates autophagy. PLoS One 5, e15394. https://doi.org/10.1371/journal.pone.0015394
  22. Levine, B. (2007). Cell biology: autophagy and cancer. Nature 446, 745-747. https://doi.org/10.1038/446745a
  23. Levine, B., and Kroemer, G. (2008). Autophagy in the pathogenesis of disease. Cell 132, 27-42. https://doi.org/10.1016/j.cell.2007.12.018
  24. Liang, X.H., Jackson, S., Seaman, M., Brown, K., Kempkes, B., Hibshoosh, H., and Levine, B. (1999). Induction of autophagy and inhibition of tumorigenesis by beclin 1. Nature 402, 672-676. https://doi.org/10.1038/45257
  25. Liu, Y.L., Yang, P.M., Shun, C.T., Wu, M.S., Weng, J.R., and Chen, C.C. (2010). Autophagy potentiates the anti-cancer effects of the histone deacetylase inhibitors in hepatocellular carcinoma. Autophagy 6, 1057-1065. https://doi.org/10.4161/auto.6.8.13365
  26. Maiuri, M.C., Zalckvar, E., Kimchi, A., and Kroemer, G. (2007). Selfeating and self-killing: crosstalk between autophagy and apoptosis. Nat. Rev. Mol. Cell Biol. 8, 741-752.
  27. Mizushima, N., Yoshimori, T., and Levine, B. (2010). Methods in mammalian autophagy research. Cell 140, 313-326. https://doi.org/10.1016/j.cell.2010.01.028
  28. Morselli, E., Galluzzi, L., Kepp, O., Vicencio, J.M., Criollo, A., Maiuri, M.C., and Kroemer, G. (2009). Anti- and pro-tumor functions of autophagy. Biochim. Biophys. Acta 1793, 1524-1532. https://doi.org/10.1016/j.bbamcr.2009.01.006
  29. Olivier, S., Close, P., Castermans, E., de Leval, L., Tabruyn, S., Chariot, A., Malaise, M., Merville, M.P., Bours, V., and Franchimont, N. (2006). Raloxifene-induced myeloma cell apoptosis: a study of nuclear factor-kappaB inhibition and gene expression signature. Mol. Pharmacol. 69, 1615-1623. https://doi.org/10.1124/mol.105.020479
  30. Powles, T. (2011). Prevention of breast cancer by newer SERMs in the future. Recent Results Cancer Res. 188, 141-145.
  31. Rossi, V., Bellastella, G., De Rosa, C., Abbondanza, C., Visconti, D., Maione, L., Chieffi, P., Della Ragione, F., Prezioso, D., De Bellis, A., et al. (2011). Raloxifene induces cell death and inhibits proliferation through multiple signaling pathways in prostate cancer cells expressing different levels of estrogen receptor alpha and beta. J. Cell. Physiol. 226, 1334-1339. https://doi.org/10.1002/jcp.22461
  32. Ryter, S.W., Cloonan, S.M., and Choi, A.M. (2013). Autophagy: a critical regulator of cellular metabolism and homeostasis. Mol. Cells 36, 7-16. https://doi.org/10.1007/s10059-013-0140-8
  33. Shibata, M.A., Morimoto, J., Shibata, E., Kurose, H., Akamatsu, K., Li, Z.L., Kusakabe, M., Ohmichi, M., and Otsuki, Y. (2010). Raloxifene inhibits tumor growth and lymph node metastasis in a xenograft model of metastatic mammary cancer. BMC Cancer 10, 566. https://doi.org/10.1186/1471-2407-10-566
  34. Takeuchi, H., Kondo, Y., Fujiwara, K., Kanzawa, T., Aoki, H., Mills, G.B., and Kondo, S. (2005). Synergistic augmentation of rapamycin- induced autophagy in malignant glioma cells by phosphatidylinositol 3-kinase/protein kinase B inhibitors. Cancer Res. 65, 3336-3346. https://doi.org/10.1158/0008-5472.CAN-04-3640
  35. Taurin, S., Allen, K.M., Scandlyn, M.J., and Rosengren, R.J. (2013). Raloxifene reduces triple-negative breast cancer tumor growth and decreases EGFR expression. Int. J. Oncol. 43, 785-792. https://doi.org/10.3892/ijo.2013.2012
  36. Wagstaff, S.C., Bowler, W.B., Gallagher, J.A., and Hipskind, R.A. (2000). Extracellular ATP activates multiple signalling pathways and potentiates growth factor-induced c-fos gene expression in MCF-7 breast cancer cells. Carcinogenesis 21, 2175-2181. https://doi.org/10.1093/carcin/21.12.2175
  37. Wang, Q., Wang, L., Feng, Y.H., Li, X., Zeng, R., and Gorodeski, G.I. (2004). P2X7 receptor-mediated apoptosis of human cervical epithelial cells. Am. J. Physiol. Cell Physiol. 287, C1349-1358. https://doi.org/10.1152/ajpcell.00256.2004
  38. White, N., and Burnstock, G. (2006). P2 receptors and cancer. Trends Pharmacol. Sci. 27, 211-217. https://doi.org/10.1016/j.tips.2006.02.004
  39. Yang, Z., and Klionsky, D.J. (2010). Mammalian autophagy: core molecular machinery and signaling regulation. Curr. Opin. Cell Biol. 22, 124-131. https://doi.org/10.1016/j.ceb.2009.11.014

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