DOI QR코드

DOI QR Code

Inhibition of cell growth and induction of apoptosis by bilobalide in FaDu human pharyngeal squamous cell carcinoma

  • Jeong, Kyung In (The Institute of Dental Science, Chosun University) ;
  • Kim, Su-Gwan (The Institute of Dental Science, Chosun University) ;
  • Go, Dae-San (The Institute of Dental Science, Chosun University) ;
  • Kim, Do Kyungm (The Institute of Dental Science, Chosun University)
  • 투고 : 2020.01.03
  • 심사 : 2020.03.18
  • 발행 : 2020.03.30

초록

Bilobalide isolated from the leaves of Ginkgo biloba has several pharmacological activities such as neuroprotective, anti-inflammatory, and anticonvulsant. However, the effect of bilobalide on cancer has not been clearly established. The main purpose of this study was to investigate the effect of bilobalide on cell growth and apoptosis induction in FaDu human pharyngeal squamous cell carcinoma. This was examined by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay, nuclear 4′,6-diamidino-2-phenylindole dihydrochloride staining, DNA fragmentation analysis, and immunoblotting. Bilobalide inhibited the growth of FaDu cells in dose- and time-dependent manners. Treatment with bilobalide resulted in nuclear condensation and DNA fragmentation in FaDu cells. Furthermore, it promoted the proteolytic cleavage of procaspase-3/-7/-8/-9 with increase in the amount of cleaved caspase-3/-7/-8/-9. Bilobalide-induced apoptosis in FaDu cells was mediated by the expression of Fas and the activation of caspase-8, caspase-3, and poly (ADP-ribose) polymerase. Immunoblotting revealed that the antiapoptotic mitochondrial protein Bcl-2 was downregulated, but the proapoptotic protein Bax was upregulated by bilobalide in FaDu cells. Bilobalide significantly increased Bax/Bcl-2 ratio. These results suggest that bilobalide inhibits cell proliferation and induces apoptosis in FaDu human pharyngeal squamous cell carcinoma via both the death receptor-mediated extrinsic apoptotic pathway and the mitochondrial-mediated intrinsic apoptotic pathway.

키워드

참고문헌

  1. Silverman S Jr, Gorsky M. Epidemiologic and demographic update in oral cancer: California and national data-1973 to 1985. J Am Dent Assoc 1990;120:495-9. doi: 10.14219/jada.archive.1990.0082.
  2. Lee JS, Seo HS, Kim SJ, Kim HJ, Kim J, Lee SH, Park YS, Park BK, Kim BS, Kim SK, Jung JY. Studies on the anticancer effect of apigenin in KB cell xenograft nude mouse model. J Life Sci 2010;20:1519-24. doi: 10.5352/JLS.2010.20.10.1519.
  3. Sacks PG. Cell, tissue and organ culture as in vitro models to study the biology of squamous cell carcinomas of the head and neck. Cancer Metastasis Rev 1996;15:27-51. doi: 10.1007/bf00049486.
  4. Todd R, Donoff RB, Wong DT. The molecular biology of oral carcinogenesis: toward a tumor progression model. J Oral Maxillofac Surg 1997;55:613-23; discussion 623-5. doi: 10.1016/S0278-2391(97)90495-X.
  5. Links M, Lewis C. Chemoprotectants: a review of their clinical pharmacology and therapeutic efficacy. Drugs 1999;57:293-308. doi: 10.2165/00003495-199957030-00003.
  6. Park DI, Lee JH, Moon SK, Kim CH, Lee YT, Cheong J, Choi BT, Choi YH. Induction of apoptosis and inhibition of telomerase activity by aqueous extract from Platycodon grandiflorum in human lung carcinoma cells. Pharmacol Res 2005;51:437-43. doi: 10.1016/j.phrs.2004.11.003.
  7. Tan ML, Sulaiman SF, Najimuddin N, Samian MR, Muhammad TS. Methanolic extract of Pereskia bleo (Kunth) DC. (Cactaceae) induces apoptosis in breast carcinoma, T47-D cell line. J Ethnopharmacol 2005;96:287-94. doi: 10.1016/j.jep.2004.09.025.
  8. Hoshino T, Hara A, Inoue M, Honda J, Imai Y, Oizumi K, Yokoyama MM. Flow cytometric measurement of NK cell cytotoxicity. J Clin Lab Immunol 1991;36:39-43.
  9. Tian Z, Chen S, Zhang Y, Huang M, Shi L, Huang F, Fong C, Yang M, Xiao P. The cytotoxicity of naturally occurring styryl lactones. Phytomedicine 2006;13:181-6. doi: 10.1016/j.phymed.2004.07.010.
  10. Green DR, Reed JC. Mitochondria and apoptosis. Science 1998;281:1309-12. doi: 10.1126/science.281.5381.1309.
  11. Hengartner MO. The biochemistry of apoptosis. Nature 2000;407:770-6. doi: 10.1038/35037710.
  12. Kaufmann SH, Hengartner MO. Programmed cell death: alive and well in the new millennium. Trends Cell Biol 2001;11:526-34. doi: 10.1016/s0962-8924(01)02173-0.
  13. Hu W, Kavanagh JJ. Anticancer therapy targeting the apoptotic pathway. Lancet Oncol 2003;4:721-9. doi: 10.1016/s1470-2045(03)01277-4.
  14. Cohen GM. Caspases: the executioners of apoptosis. Biochem J 1997;326(Pt 1):1-16. doi: 10.1042/bj3260001.
  15. Kaufmann SH, Earnshaw WC. Induction of apoptosis by cancer chemotherapy. Exp Cell Res 2000;256:42-9. doi: 10.1006/excr.2000.4838.
  16. Reed JC. Apoptosis-regulating proteins as targets for drug discovery. Trends Mol Med 2001;7:314-9. doi: 10.1016/s1471-4914(01)02026-3.
  17. Shi C, Wu F, Yew DT, Xu J, Zhu Y. Bilobalide prevents apoptosis through activation of the PI3K/Akt pathway in SH-SY5Y cells. Apoptosis 2010;15:715-27. doi: 10.1007/s10495-010-0492-x.
  18. Suzuki R, Kohno H, Sugie S, Sasaki K, Yoshimura T, Wada K, Tanaka T. Preventive effects of extract of leaves of ginkgo (Ginkgo biloba) and its component bilobalide on azoxymethane-induced colonic aberrant crypt foci in rats. Cancer Lett 2004;210:159-69. doi: 10.1016/j.canlet.2004.01.034.
  19. Jiang M, Li J, Peng Q, Liu Y, Liu W, Luo C, Peng J, Li J, Yung KK, Mo Z. Neuroprotective effects of bilobalide on cerebral ischemia and reperfusion injury are associated with inhibition of pro-inflammatory mediator production and down-regulation of JNK1/2 and p38 MAPK activation. J Neuroinflammation 2014;11:167. doi: 10.1186/s12974-014-0167-6.
  20. Priyanka A, Nisha VM, Anusree SS, Raghu KG. Bilobalide attenuates hypoxia induced oxidative stress, inflammation, and mitochondrial dysfunctions in 3T3-L1 adipocytes via its antioxidant potential. Free Radic Res 2014;48:1206-17. doi: 10.3109/10715762.2014.945442.
  21. Yang SS, Kim SG, Park BS, Go DS, Yu SK, Kim CS, Kim J, Kim DK. Effect of ${\beta}$-carotene on cell growth inhibition of KB human oral cancer cells. Int J Oral Biol 2016;41:105-11. doi: 10.11620/IJOB.2016.41.3.105.
  22. Cho JH, Kim SG, Park BS, Go DS, Park JC, Kim DK. MicroRNA-27 promotes odontoblast differentiation via Wnt1 signaling. Int J Oral Biol 2015;40:197-204. doi: 10.11620/IJOB.2015.40.4.197.
  23. Herrnring C, Reimer T, Jeschke U, Makovitzky J, Kruger K, Gerber B, Kabelitz D, Friese K. Expression of the apoptosisinducing ligands FasL and TRAIL in malignant and benign human breast tumors. Histochem Cell Biol 2000;113:189-94. doi: 10.1007/s004180050438.
  24. Li HJ, Wang CY, Mi Y, Du CG, Cao GF, Sun XC, Liu DJ, Shorgan B. FasL-induced apoptosis in bovine oocytes via the Bax signal. Theriogenology 2013;80:248-55. doi: 10.1016/j.theriogenology.2013.04.002.
  25. Datta R, Kojima H, Yoshida K, Kufe D. Caspase-3-mediated cleavage of protein kinase C theta in induction of apoptosis. J Biol Chem 1997;272:20317-20. doi: 10.1074/jbc.272.33.20317.
  26. Liu X, Zou H, Slaughter C, Wang X. DFF, a heterodimeric protein that functions downstream of caspase-3 to trigger DNA fragmentation during apoptosis. Cell 1997;89:175-84. doi: 10.1016/s0092-8674(00)80197-x.
  27. Yang SJ, Lee SA, Park MG, Kim JS, Yu SK, Kim CS, Kim JS, Kim SG, Oh JS, Kim HJ, Chun HS, Kim YH, Kim DK. Induction of apoptosis by diphenyldifluoroketone in osteogenic sarcoma cells is associated with activation of caspases. Oncol Rep 2014;31:2286-92. doi: 10.3892/or.2014.3066.
  28. Kluck RM, Bossy-Wetzel E, Green DR, Newmeyer DD. The release of cytochrome c from mitochondria: a primary site for Bcl-2 regulation of apoptosis. Science 1997;275:1132-6. doi: 10.1126/science.275.5303.1132.
  29. Kluck RM, Esposti MD, Perkins G, Renken C, Kuwana T, Bossy-Wetzel E, Goldberg M, Allen T, Barber MJ, Green DR, Newmeyer DD. The pro-apoptotic proteins, Bid and Bax, cause a limited permeabilization of the mitochondrial outer membrane that is enhanced by cytosol. J Cell Biol 1999;147:809-22. doi: 10.1083/jcb.147.4.809.
  30. Yu CS, Huang AC, Lai KC, Huang YP, Lin MW, Yang JS, Chung JG. Diallyl trisulfide induces apoptosis in human primary colorectal cancer cells. Oncol Rep 2012;28:949-54. doi: 10.3892/or.2012.1882.
  31. Oltvai ZN, Milliman CL, Korsmeyer SJ. Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programmed cell death. Cell 1993;74:609-19. doi: 10.1016/0092-8674(93)90509-o.