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Betulinic Acid Induces Apoptosis in Humam Mucoepidermoid Carcinoma Cells Through Regulating Specificity Protein 1 and Its Downstream Molecule, Survivin

  • Lee, Jung-Eun (Department of Oral Pathology, School of Dentistry, Institute of Oral Bioscience, Brain Korea 21, Chonbuk National University) ;
  • Jung, Ji-Youn (Department of Companion and Laboratory Animal Science, Kongju National University) ;
  • Yoo, Hyun-Ju (Department of Oral Pathology, School of Dentistry, Institute of Oral Bioscience, Brain Korea 21, Chonbuk National University) ;
  • Cho, Sung-Dae (Department of Oral Pathology, School of Dentistry, Institute of Oral Bioscience, Brain Korea 21, Chonbuk National University)
  • Received : 2013.02.28
  • Accepted : 2013.07.31
  • Published : 2013.09.30

Abstract

High-grade mucoepidermoid carcinomas (MECs) have difficulty in cure and 5-year survival rate is quiet low. Therefore, we need new therapeutic agents and molecular targets. Betulinic acid (BA) is one of the materials which is easily found in the world and shows tumor-suppress effects in various tumor types. In addition, many kinds of normal tissues have a resistance to BA treatment. In this study, we investigated the anti-proliferative activity of BA and its molecular targets in MC-3 human MEC cells using western blot analysis and DAPI staining. BA inhibited cell viability and induced apoptosis in MC-3 cells. It affected Specificity protein 1 (Sp1) and its downstream molecule, survivin whereas it did not affect myeloid cell leukemia-1 (Mcl-1). Therefore, we suggest that BA can be a potential anti-cancer drug candidate regulating Sp 1 and survivin to exert apoptotic cell death.

악성도가 높은 점액표피양 암종은 치료가 어렵고 5년 생존률이 매우 낮다. 따라서, 새로운 치료 물질과 분자표적을 찾는 것이 필요하다. Betulinic acid (BA)는 세계적으로 쉽게 얻을 수 있는 물질인 동시에 여러 종류의 종양에서 항암효과를 보인다. 또한 여러 정상 조직은 BA에 저항성을 보인다. 이 연구에서는 BA의 증식억제 효능과 MC-3 세포주에서의 분자 표적을 확인하고자 하였다. BA는 MC-3 세포주에서 세포 생존을 저해하였고 세포사멸을 유도하였다. BA는 Sp 1과 그의 하향 분자 표적인 survivin에 영향을 주었으나, 다른 하향 분자 표적인 Mcl-1에서는 유의한 변화를 일으키지 못하였다. 따라서, BA는 Sp1과 survivin을 조절하여 세포사멸을 일으키는 잠재적인 항암제 후보가 될 수 있을 것이라 사료된다.

Keywords

References

  1. Enlund, F., et al., Altered Notch signaling resulting from expression of a WAMTP1-MAML2 gene fusion in mucoepidermoid carcinomas and benign Warthin's tumors. Exp Cell Res, 292(1), 21-8 (2004). https://doi.org/10.1016/j.yexcr.2003.09.007
  2. Behboudi, A., et al., Molecular classification of mucoepidermoid carcinomas-prognostic significance of the MECT1-MAML2 fusion oncogene. Genes Chromosomes Cancer, 45(5), 470-81 (2006). https://doi.org/10.1002/gcc.20306
  3. Kaye, F.J., Mutation-associated fusion cancer genes in solid tumors. Mol Cancer Ther, 8(6), 1399-408 (2009). https://doi.org/10.1158/1535-7163.MCT-09-0135
  4. Nance, M.A., et al., Treatment and survival outcomes based on histologic grading in patients with head and neck mucoepidermoid carcinoma. Cancer,. 113(8), 2082-9 (2008). https://doi.org/10.1002/cncr.23825
  5. Newman, D.J., G.M. Cragg, and K.M. Snader, Natural products as sources of new drugs over the period 1981-2002. J Nat Prod, 66(7), 1022-37 (2003). https://doi.org/10.1021/np030096l
  6. Paduch, R., et al., Terpenes: substances useful in human healthcare. Arch Immunol Ther Exp (Warsz), 55(5), 315-27 (2007). https://doi.org/10.1007/s00005-007-0039-1
  7. Liby, K.T., M.M. Yore, and M.B. Sporn, Triterpenoids and rexinoids as multifunctional agents for the prevention and treatment of cancer. Nat Rev Cancer, 7(5), 357-69 (2007). https://doi.org/10.1038/nrc2129
  8. Alakurtti, S., et al., Pharmacological properties of the ubiquitous natural product betulin. Eur J Pharm Sci, 29(1), 1-13 (2006). https://doi.org/10.1016/j.ejps.2006.04.006
  9. Fulda, S. and G. Kroemer, Targeting mitochondrial apoptosis by betulinic acid in human cancers. Drug Discov Today, 14(17-18), 885-90 (2009). https://doi.org/10.1016/j.drudis.2009.05.015
  10. Santos, R.C., et al., New betulinic acid derivatives induce potent and selective antiproliferative activity through cell cycle arrest at the S phase and caspase dependent apoptosis in human cancer cells. Biochimie, 93(6), 1065-75 (2011). https://doi.org/10.1016/j.biochi.2011.02.014
  11. Fulda, S., Betulinic Acid for cancer treatment and prevention. Int J Mol Sci, 9(6), 1096-107 (2008). https://doi.org/10.3390/ijms9061096
  12. Thurnher, D., et al., Betulinic acid: a new cytotoxic compound against malignant head and neck cancer cells. Head Neck, 25(9), 732-40 (2003). https://doi.org/10.1002/hed.10231
  13. Chang, W.C. and J.J. Hung, Functional role of post-translational modifications of Sp1 in tumorigenesis. J Biomed Sci, 19, 94 (2012). https://doi.org/10.1186/1423-0127-19-94
  14. Saxena, A., et al., Short nucleotide polymorphic insertions in the MCL-1 promoter affect gene expression. Cancer Lett, 251(1), 114-31 (2007). https://doi.org/10.1016/j.canlet.2006.11.007
  15. Shin, J.A., et al., Apoptotic effect of Polygonum Cuspidatum in oral cancer cells through the regulation of specificity protein 1. Oral Dis, 17(2), 162-70 (2011). https://doi.org/10.1111/j.1601-0825.2010.01710.x
  16. McHugh, C.H., et al., Prognostic factors in mucoepidermoid carcinoma of the salivary glands. Cancer, 118(16), 3928-36 (2012). https://doi.org/10.1002/cncr.26697
  17. Chiosea, S.I., et al., Mucoepidermoid carcinoma of upper aerodigestive tract: clinicopathologic study of 78 cases with immunohistochemical analysis of Dicer expression. Virchows Arch, 452(6), 629-35 (2008). https://doi.org/10.1007/s00428-007-0574-5
  18. Cichewicz, R.H. and S.A. Kouzi, Chemistry, biological activity, and chemotherapeutic potential of betulinic acid for the prevention and treatment of cancer and HIV infection. Med Res Rev, 24(1), 90-114 (2004). https://doi.org/10.1002/med.10053
  19. Galluzzi, L., et al., Mitochondria as therapeutic targets for cancer chemotherapy. Oncogene, 25(34), 4812-30 (2006). https://doi.org/10.1038/sj.onc.1209598
  20. Zuco, V., et al., Selective cytotoxicity of betulinic acid on tumor cell lines, but not on normal cells. Cancer Lett, 175(1), 17-25 (2002). https://doi.org/10.1016/S0304-3835(01)00718-2
  21. Fulda, S., et al., Betulinic acid: a new chemotherapeutic agent in the treatment of neuroectodermal tumors. Klin Padiatr, 211(4), 319-22 (1999). https://doi.org/10.1055/s-2008-1043808
  22. Suske, G., The Sp-family of transcription factors. Gene, 238(2), 291-300 (1999). https://doi.org/10.1016/S0378-1119(99)00357-1
  23. Black, A.R., J.D. Black, and J. Azizkhan-Clifford, Sp1 and kruppel-like factor family of transcription factors in cell growth regulation and cancer. J Cell Physiol, 188(2), 143-60 (2001). https://doi.org/10.1002/jcp.1111
  24. Abdelrahim, M., et al., Small inhibitory RNA duplexes for Sp1 mRNA block basal and estrogen-induced gene expression and cell cycle progression in MCF-7 breast cancer cells. J Biol Chem, 277(32), 28815-22 (2002). https://doi.org/10.1074/jbc.M203828200
  25. Abdelrahim, M. and S. Safe, Cyclooxygenase-2 inhibitors decrease vascular endothelial growth factor expression in colon cancer cells by enhanced degradation of Sp1 and Sp4 proteins. Mol Pharmacol, 68(2), 317-29 (2005).
  26. Yuan, P., et al., Therapeutic inhibition of Sp1 expression in growing tumors by mithramycin a correlates directly with potent antiangiogenic effects on human pancreatic cancer. Cancer, 110(12), 2682-90 (2007). https://doi.org/10.1002/cncr.23092
  27. Ishibashi, H., et al., Sp1 decoy transfected to carcinoma cells suppresses the expression of vascular endothelial growth factor, transforming growth factor beta1, and tissue factor and also cell growth and invasion activities. Cancer Res, 60(22), 6531-6 (2000).
  28. Lou, Z., et al., Down-regulation of overexpressed sp1 protein in human fibrosarcoma cell lines inhibits tumor formation. Cancer Res, 65(3), 1007-17 (2005).
  29. Adrian, G.S., et al., YY1 and Sp1 transcription factors bind the human transferrin gene in an age-related manner. J Gerontol A Biol Sci Med Sci, 51(1), B66-75 (1996).
  30. Oh, J.E., J.A. Han, and E.S. Hwang, Downregulation of transcription factor, Sp1, during cellular senescence. Biochem Biophys Res Commun, 353(1), 86-91 (2007). https://doi.org/10.1016/j.bbrc.2006.11.118
  31. Deveraux, Q.L. and J.C. Reed, IAP family proteins--suppressors of apoptosis. Genes Dev, 13(3), 239-52 (1999). https://doi.org/10.1101/gad.13.3.239
  32. Li, F., Survivin study: what is the next wave? J Cell Physiol, 197(1), 8-29 (2003). https://doi.org/10.1002/jcp.10327
  33. Altieri, D.C., Survivin in apoptosis control and cell cycle regulation in cancer. Prog Cell Cycle Res, 5, 447-52 (2003).
  34. Li, F. and M.G. Brattain, Role of the Survivin gene in pathophysiology. Am J Pathol, 169(1), 1-11 (2006). https://doi.org/10.2353/ajpath.2006.060121
  35. Dasgupta, P., et al., Nicotine inhibits apoptosis induced by chemotherapeutic drugs by up-regulating XIAP and survivin. Proc Natl Acad Sci U S A, 103(16), 6332-7 (2006). https://doi.org/10.1073/pnas.0509313103
  36. Virrey, J.J., et al., Increased survivin expression confers chemoresistance to tumor-associated endothelial cells. Am J Pathol, 173(2), 575-85 (2008). https://doi.org/10.2353/ajpath.2008.071079
  37. Li, F., Role of survivin and its splice variants in tumorigenesis. Br J Cancer, 92(2), 212-6 (2005).
  38. Zhou, P., et al., Mcl-1, a Bcl-2 family member, delays the death of hematopoietic cells under a variety of apoptosisinducing conditions. Blood, 89(2), 630-43 (1997).
  39. Moulding, D.A., et al., Apoptosis is rapidly triggered by antisense depletion of MCL-1 in differentiating U937 cells. Blood, 96(5), 1756-63 (2000).
  40. Wei, L.H., et al., The anti-apoptotic role of interleukin-6 in human cervical cancer is mediated by up-regulation of Mcl-1 through a PI 3-K/Akt pathway. Oncogene, 20(41), 5799-809 (2001). https://doi.org/10.1038/sj.onc.1204733