International Journal of Oral Biology

The Korean Academy of Oral Biology (대한구강생물학회)
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Apoptotic Effects of Co-Treatment with a Chios Gum Mastic and Eugenol on G361 Human Melanoma Cells

  • Jo, Jae-Beom (Department of Oral Anatomy, School of Dentistry, Pusan National University) ;
  • Oh, Sang-Hun (Department of Oral Anatomy, School of Dentistry, Pusan National University) ;
  • Kim, In-Ryoung (Department of Oral Anatomy, School of Dentistry, Pusan National University) ;
  • Kim, Gyoo-Cheon (Department of Oral Anatomy, School of Dentistry, Pusan National University) ;
  • Kwak, Hyun-Ho (Department of Oral Anatomy, School of Dentistry, Pusan National University) ;
  • Park, Bong-Soo (Department of Oral Anatomy, School of Dentistry, Pusan National University)
  • Received : 2013.07.09
  • Accepted : 2013.08.22
  • Published : 2013.09.30
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Abstract

We investigated the synergistic apoptotic effects of co-treatments with Chios gum mastic (CGM) and eugenol on G361 human melanoma cells. An MTT assay was conducted to investigate whether this co-treatment efficiently reduces the viability of G361 cells compared with each single treatment. The induction and augmentation of apoptosis were confirmed by DNA electrophoresis, Hoechst staining, and analyses of DNA hypoploidy. Western blot analysis and immunofluorescent staining were also performed to evaluate expression and translocation of apoptosis-related proteins following CGM and eugenol co-treatment. Proteasome activity and mitochondrial membrane potential (MMP) changes were also assayed.The results indicated that the co-treatment of CGM and eugenol induces multiple pathways and processes associated with an apoptotic response in G361 cells. These include nuclear condensation, DNA fragmentation, a reduction in MMP and proteasome activity, an increase of Bax and decrease of Bcl-2, a decreased DNA content, cytochrome c release into the cytosol, the translocation of AIF and DFF40 (CAD) into the nucleus, and the activation of caspase-9, caspase-7, caspase-3, PARP and DFF45 (ICAD). In contrast, separate treatments of $40{\mu}g/ml$ CGM or $300{\mu}M$ eugenol for 24 hours did not induce apoptosis. Our present data thus suggest that a combination therapy of CGM and eugenol is a potential treatment strategy for human melanoma.

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References

  1. Williams GT, Programmed cell death: apoptosis and oncogenesis. Cell, 1991;65:1097-8. https://doi.org/10.1016/0092-8674(91)90002-G
  2. Yuan J, Evolutionary conservation of a genetic pathway of programmed cell death. J Cell Biochem, 1996;60:4-11. https://doi.org/10.1002/(SICI)1097-4644(19960101)60:1<4::AID-JCB2>3.0.CO;2-1
  3. Susin SA, HK Lorenzo, N Zamzami, I Marzo, BE Snow, GM Brothers, J Mangion, E Jacotot, P Costantini, M Loeffler, N Larochette, DR Goodlett, R Aebersold, DP Siderovski, JM Penninger, and G Kroemer, Molecular characterization of mitochondrial apoptosis-inducing factor. Nature, 1999;397:441-6. https://doi.org/10.1038/17135
  4. Orlowski RZ, The role of the ubiquitin-proteasome pathway in apoptosis. Cell Death Differ, 1999;6:303-13. https://doi.org/10.1038/sj.cdd.4400505
  5. Shibuya H, Y Kato, M Saito, T Isobe, R Tsuboi, M Koga, H Toyota, and J Mizuguchi, Induction of apoptosis and/or necrosis following exposure to antitumour agents in a melanoma cell line, probably through modulation of Bcl-2 family proteins. Melanoma Res, 2003;13:457-64. https://doi.org/10.1097/00008390-200310000-00004
  6. Balan KV, J Prince, Z Han, K Dimas, M Cladaras, JH Wyche, NM Sitaras, and P Pantazis, Antiproliferative activity and induction of apoptosis in human colon cancer cells treated in vitro with constituents of a product derived from Pistacia lentiscus L. var. chia. Phytomedicine, 2007;14:263-72. https://doi.org/10.1016/j.phymed.2006.03.009
  7. He ML, A Li, CS Xu, SL Wang, MJ Zhang, H Gu, YQ Yang, and HH Tao, Mechanisms of antiprostate cancer by gum mastic: NF-kappaB signal as target. Acta Pharmacol Sin, 2007;28:446-52. https://doi.org/10.1111/j.1745-7254.2007.00536.x
  8. Markowitz K, M Moynihan, M Liu, and S Kim, Biologic properties of eugenol and zinc oxide-eugenol. A clinically oriented review. Oral Surg Oral Med Oral Pathol, 1992;73:729-37. https://doi.org/10.1016/0030-4220(92)90020-Q
  9. Suzuki Y, K Sugiyama, and H Furuta, Eugenol-mediated superoxide generation and cytotoxicity in guinea pig neutrophils. Jpn J Pharmacol, 1985;39:381-6. https://doi.org/10.1254/jjp.39.381
  10. Wright SE, DA Baron, and JE Heffner, Intravenous eugenol causes hemorrhagic lung edema in rats: proposed oxidant mechanisms. J Lab Clin Med, 1995;125:257-64.
  11. Satoh K, H Sakagami, I Yokoe, M Kochi, and S Fujisawa, Interaction between eugenol-related compounds and radicals. Anticancer Res, 1998;18:425-8.
  12. Adhami VM, A Malik, N Zaman, S Sarfaraz, IA Siddiqui, DN Syed, F Afaq, FS Pasha, M Saleem, and H Mukhtar, Combined inhibitory effects of green tea polyphenols and selective cyclooxygenase-2 inhibitors on the growth of human prostate cancer cells both in vitro and in vivo. Clin Cancer Res, 2007;13:1611-9. https://doi.org/10.1158/1078-0432.CCR-06-2269
  13. Mai Z, GL Blackburn, and JR Zhou, Genistein sensitizes inhibitory effect of tamoxifen on the growth of estrogen receptor-positive and HER2-overexpressing human breast cancer cells. Mol Carcinog, 2007;46:534-42. https://doi.org/10.1002/mc.20300
  14. Song MQ, JS Zhu, JL Chen, L Wang, W Da, L Zhu, and WP Zhang, Synergistic effect of oxymatrine and angiogenesis inhibitor NM-3 on modulating apoptosis in human gastric cancer cells. World J Gastroenterol, 2007;13:1788-93.
  15. Vinall RL, K Hwa, P Ghosh, CX Pan, PN Lara, Jr., and RW de Vere White, Combination treatment of prostate cancer cell lines with bioactive soy isoflavones and perifosine causes increased growth arrest and/or apoptosis. Clin Cancer Res, 2007;13:6204-16. https://doi.org/10.1158/1078-0432.CCR-07-0600
  16. Lee SH, JK Ryu, KY Lee, SM Woo, JK Park, JW Yoo, YT Kim, and YB Yoon, Enhanced anti-tumor effect of combination therapy with gemcitabine and apigenin in pancreatic cancer. Cancer Lett, 2008;259:39-49. https://doi.org/10.1016/j.canlet.2007.09.015
  17. Zhu J, L Miao, C Xu, Y Wang, and T Chen, [Analysis of 822 infants with limb reduction defect in China]. Hua Xi Yi Ke Da Xue Xue Bao, 1996;27:400-3.
  18. Kim HM, EH Lee, SH Hong, HJ Song, MK Shin, SH Kim, and TY Shin, Effect of Syzygium aromaticum extract on immediate hypersensitivity in rats. J Ethnopharmacol, 1998;60:125-31. https://doi.org/10.1016/S0378-8741(97)00143-8
  19. Kim HM, JM Yi, and KS Lim, Magnoliae flos inhibits mast cell-dependent immediate-type allergic reactions. Pharmacol Res, 1999;39:107-11. https://doi.org/10.1006/phrs.1998.0414
  20. Park HI, MH Jeong, YJ Lim, BS Park, GC Kim, YM Lee, HM Kim, KS Yoo, and YH Yoo, Szygium aromaticum (L.) Merr. Et Perry (Myrtaceae) flower bud induces apoptosis of p815 mastocytoma cell line. Life Sci, 2001;69:553-66. https://doi.org/10.1016/S0024-3205(01)01140-7
  21. Na HJ, HJ Jeong, H Bae, YB Kim, ST Park, YG Yun, and HM Kim, Tongkyutang inhibits mast cell-dependent allergic reactions and inflammatory cytokines secretion. Clin Chim Acta, 2002;319:35-41. https://doi.org/10.1016/S0009-8981(02)00011-6
  22. Kim JM, HR Bae, BS Park, JM Lee, HB Ahn, JH Rho, KW Yoo, WC Park, SH Rho, HS Yoon, and YH Yoo, Early mitochondrial hyperpolarization and intracellular alkalinization in lactacystin-induced apoptosis of retinal pigment epithelial cells. J Pharmacol Exp Ther, 2003;305:474-81. https://doi.org/10.1124/jpet.102.047811
  23. Im SJ, KN Kim, YG Yun, JC Lee, YJ Mun, JH Kim, and WH Woo, Effect of Radix Ginseng and Radix Trichosanthis on the melanogenesis. Biol Pharm Bull, 2003;26:849-53. https://doi.org/10.1248/bpb.26.849
  24. Dattner AM, Herbal and complementary medicine in dermatology. Dermatol Clin, 2004;22:325-32, vii.
  25. Lee KJ, JY Kim, JH Choi, HG Kim, YC Chung, SH Roh, and HG Jeong, Inhibition of tumor invasion and metastasis by aqueous extract of the radix of Platycodon grandiflorum. Food Chem Toxicol, 2006;44:1890-6. https://doi.org/10.1016/j.fct.2006.06.009
  26. IARC, Monographs on the Evaluation of Carcinogenic Risks to Humans. Allyl Compounds, Aldehydes, Epoxides and Peroxides 1985;36:75.
  27. Zhu YP, Materia Medica: Cemistry, Pharmacology and Application, 1st edi., Amsterdam. Harwood Academic Publishers, 1996;1-3.
  28. Kim HM, EH Lee, CY Kim, JG Chung, SH Kim, JP Lim, and TY Shin, Antianaphylactic properties of eugenol. Pharmacol Res, 1997;36:475-80. https://doi.org/10.1006/phrs.1997.0256
  29. Shin BK, EH Lee, and HM Kim, Suppression of Lhistidine decarboxylase mRNA expression by methyleugenol. Biochem Biophys Res Commun, 1997;232:188-91. https://doi.org/10.1006/bbrc.1997.6260
  30. Al-Habbal MJ, Z Al-Habbal, and FU Huwez, A doubleblind controlled clinical trial of mastic and placebo in the treatment of duodenal ulcer. Clin Exp Pharmacol Physiol, 1984;11:541-4. https://doi.org/10.1111/j.1440-1681.1984.tb00864.x
  31. Al-Said MS, AM Ageel, NS Parmar, and M Tariq, Evaluation of mastic, a crude drug obtained from Pistacia lentiscus for gastric and duodenal anti-ulcer activity. J Ethnopharmacol, 1986;15:271-8. https://doi.org/10.1016/0378-8741(86)90165-0
  32. Huwez FU and MJ Al-Habbal, Mastic in treatment of benign gastric ulcers. Gastroenterol Jpn, 1986;21:273-4.
  33. Correction: Mastic Gum Kills Helicobacter pylori. N Engl J Med, 1999;340:576.
  34. Aksoy A, N Duran, and F Koksal, In vitro and in vivo antimicrobial effects of mastic chewing gum against Streptococcus mutans and mutans streptococci. Arch Oral Biol, 2006;51:476-81. https://doi.org/10.1016/j.archoralbio.2005.11.003
  35. Park JH, Kim GC, Kwak HH, Kim IR, Lee SE, Chung J, Park HR, Shin SH, Choi SH, Kim CH, Nam CO, Park BS, Chios Gum Mastic Induces Cell Cycle Arrest and Apoptosis in YD9 Human Oral Squamous Carcinoma Cells. Korean J Phys Anthropol 2008;21:55-68.
  36. Drexler HC, W Risau, and MA Konerding, Inhibition of proteasome function induces programmed cell death in proliferating endothelial cells. FASEB J, 2000;14:65-77.
  37. Kroemer G, N Zamzami, and SA Susin, Mitochondrial control of apoptosis. Immunol Today, 1997;18:44-51. https://doi.org/10.1016/S0167-5699(97)80014-X
  38. Green DR and JC Reed, Mitochondria and apoptosis. Science, 1998;281:1309-12. https://doi.org/10.1126/science.281.5381.1309
  39. Golab J, T Stoklosa, A Czajka, A Dabrowska, M Jakobisiak, R Zagozdzon, C Wojcik, M Marczak, and S Wilk, Synergistic antitumor effects of a selective proteasome inhibitor and TNF in mice. Anticancer Res, 2000;20:1717-21.
  40. Wagenknecht B, M Hermisson, P Groscurth, P Liston, PH Krammer, and M Weller, Proteasome inhibitor-induced apoptosis of glioma cells involves the processing of multiple caspases and cytochrome c release. J Neurochem, 2000;75:2288-97.
  41. Marshansky V, X Wang, R Bertrand, H Luo, W Duguid, G Chinnadurai, N Kanaan, MD Vu, and J Wu, Proteasomes modulate balance among proapoptotic and antiapoptotic Bcl-2 family members and compromise functioning of the electron transport chain in leukemic cells. J Immunol, 2001;166:3130-42.
  42. Daugas E, SA Susin, N Zamzami, KF Ferri, T Irinopoulou, N Larochette, MC Prevost, B Leber, D Andrews, J Penninger, and G Kroemer, Mitochondrio-nuclear translocation of AIF in apoptosis and necrosis. FASEB J, 2000;14:729-39.
  43. Acehan D, X Jiang, DG Morgan, JE Heuser, X Wang, and CW Akey, Three-dimensional structure of the apoptosome: implications for assembly, procaspase-9 binding, and activation. Mol Cell, 2002;9:423-32. https://doi.org/10.1016/S1097-2765(02)00442-2
  44. Gross A, JM McDonnell, and SJ Korsmeyer, BCL-2 family members and the mitochondria in apoptosis. Genes Dev, 1999;13:1899-911. https://doi.org/10.1101/gad.13.15.1899
  45. Porter AG, Protein translocation in apoptosis. Trends Cell Biol, 1999;9:394-401. https://doi.org/10.1016/S0962-8924(99)01624-4
  46. Cheng AC, CB Jian, YT Huang, CS Lai, PC Hsu, and MH Pan, Induction of apoptosis by Uncaria tomentosa through reactive oxygen species production, cytochrome c release, and caspases activation in human leukemia cells. Food Chem Toxicol, 2007;45:2206-18. https://doi.org/10.1016/j.fct.2007.05.016