Anti-cancer Activity of Anthricin through Caspase-dependent Apoptosis in Human Hypopharyngeal Squamous Carcinoma Cell

  • Kim, Won Gi (Department of Oral and Maxillofacial Surgery, College of Dentistry, Chosun University) ;
  • Lee, Seul Ah (Department of Oral Biochemistry, College of Dentistry, Chosun University) ;
  • Moon, Sung Min (Department of Oral Biochemistry, College of Dentistry, Chosun University) ;
  • Kim, Jin-Soo (Oral Biology Research Institute, Chosun University) ;
  • Kim, Su-Gwan (Oral Biology Research Institute, Chosun University) ;
  • Shin, Yong Kook (School of integrated Oriental Medical Bioscience, Semyung University) ;
  • Kim, Do Kyung (Oral Biology Research Institute, Chosun University) ;
  • Kim, Chun Sung (Department of Oral Biochemistry, College of Dentistry, Chosun University)
  • Received : 2016.10.26
  • Accepted : 2016.11.12
  • Published : 2016.12.31


Anthricin (Deoxypodophyllotoxin), a naturally occurring flavolignan, has well known anti-cancer properties in several cancer cells, such as prostate cancer, cervical carcinoma and pancreatic cancer. However, the effects of Anthricin are currently unknown in oral cancer. We examined the anticancer effect and mechanism of action of Anthricin in human FaDu hypopharyngeal squamous carcinoma cells. Our data showed that Anthricin inhibits cell viability in a dose- and time-dependent manner ($IC_{50}$ 50 nM) in the MTT assay and Live & Dead assay. In addition, Anthricin treated FaDu cells showed marked apoptosis by DAPI stain and FACS. Furthermore, Anthricin activates anti-apoptotic factors such as caspase-3, -9 and poly (ADP-ribose) polymerase (PARP), suggesting that caspase-mediated pathways are involved in Anthricin- induced apoptosis. Anthricin treatment also leads to accumulation of the pro-apoptotic factor Bax, followed by inhibition of cell growth. Taken together, these results indicate that Anthricn-induced cell death of human FaDu hypopharyngeal squamous carcinoma cells is mediated by mitochondrial-dependent apoptotic pathway. In summary, our findings provide a framework for further exploration on Anthricin as a novel chemotherapeutic drug for human oral cancer.


Supported by : 조선대학교


  1. Greenlee RT, Hill-Harmon MB, Murray T, Thun M. Cancer statistics. 2001. CA Cancer J Clin. 2001;51:15-36.
  2. Maggioni D, Biffi L, Nicolini G, Garavello W. Flavonoids in oral cancer prevention and therapy. Eur J Cancer Prev. 2015;24:517-528. doi:10.1097/CEJ.0000000000000109.
  3. Hopper C, Kubler A, Lewis H, Tan IB, Putnam G. mTHPC-mediated photodynamic therapy for early oral squamous cell carcinoma. Int J Cancer. 2004;111:138-146.
  4. Synytsya A, Mickova K, Synytsya A, Jablonsky I, Spevacek J, Erban V. Glucans from fruit bodies of cultivated mushrooms Pleurotus ostreatus and Pleurotus eryngii structure and potential prebiotic activity. Carbohydr Polym. doi:10.1016/j.carbol.2008.11.021.
  5. Yong Y, Shin SY, Lee YH, Lim Y. Antitumor activity of deoxypodophyllotoxin isolated from Anthriscus sylvestris: induction of G2/M cell cycle arrest and caspase-dependent apoptosis. Bioorg Med Chem Lett. 2009;19:4367-4371. doi:10.1016/j.bmcl.2009.05.093.
  6. Ikeda R, Nagao T, Okabe H, Nakano Y, Matsunaga H, Katano M, Mori M. Antiproliferative constituents in umbelliferae plants. III. Constituents in the root and the ground part of Anthriscus sylvestris Hoffm. Chem Pharm Bull (Tokyo). 1998;46:871-874.
  7. Kim Y, Kim SB, You YJ, Ahn BZ. Deoxypodophyllotoxin; the cytotoxic and antiangiogenic component from Pulsatilla koreana. Planta Medica. 2002;68:271-274. doi:10.1055/s-2002-23140.
  8. Lin CX, Son MJ, Ju HK, Moon TC, Lee E, Kim SH, Kim MJ, Son JK, Lee SH, Chang HW. Deoxypodophyllotoxin, a naturally occurring lignan, inhibits the passive cutaneous anaphylaxis reaction. Planta Medica. 2004;70:474-476. doi:10.1055/s-2004-818981.
  9. Kim KY, Cho HJ, Yu SN, Kim SH, Yu HS, Park YM, Mirkheshti N, Kim SY, Song CS, Chatterjee B, Ahn SC. Interplay of reactive oxygen species, intracellular Ca(2+) surge and loss of mitochondrial homeostasis in the apoptotic ablation of prostate cancer cells by deoxypodophyllotoxin. J Cell Biochem. 2012;114:1124-1134. doi:10.1002/jcb.24455.
  10. Shin SY, Yong Y, Kim CG, Lee YH, Lim Y. Deoxypodophyllotoxin induces G2/M cell cycle arrest and apoptosis in HeLa cells. Cancer Lett. 2010;287:231-239. doi:10.1016/j.canlet.2009.06.019.
  11. Jeong GS, Kwon OK, Park BY, Oh SR, Ahn KS, Chang MJ, Oh WK, Kimg JC, Min BS, Kim YC, Lee HK. Lignans and coumarins from the roots of Anthriscus sylvestris and their increase of caspase-3 activity in HL-60 cells. Biol Pharm Bull. 2007;30:1340-1343.
  12. Moon KS, Ji JY, Cho YJ, Lee JH, Choi MS, Kim EE. Therapeutic effects of SB natural anticancer drug in 50 patients with stage IV pancreatic cancer. J Cancer Treatment and Research. 2015;3:42-46. doi:10.11648/j.jctr.20150303.14.
  13. Wang Y, Wang B, Guerrram M, Sun L, Shi W, Tian C, Zhu X, Jiang Z, Zhang L. Deoxypodophyllotoxin suppresses turmor vasculature in HUVECs by promoting cytoskeleton remodeling through LKB-AMPK dependent Rho A activation. Oncotarget. 2015;6:29497-29512. doi:10.18632/oncotarget.4985.
  14. Cheng YL, Lee SC, Lin SZ, Chang WL, Chen YL, Tsai NM, Liu YC, Tzao C, Yu DS, Harn HJ. Anti-proliferative activity of Bupleurum scrozonerifolium in A549 human lung cancer cells in vitro and in vivo. Cancer Lett. 2005;222:183-193. doi:10.1016/j.canlet.2004.10.015.
  15. 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 grandiforum in human lung carcinama cells. Pharmacol Res. 2005;51:437-443. doi:10.1016/j.phrs.2004.11.00.
  16. Lee SH, Son MJ, Ju HK, Lin CX, Moon TC, Choi HG, Son JK, Chang HW. Dual inhibition of cyclooxygenase-2 and 5-lipoxynenase by deoxypodophyllotoxin in mouse bone marrow-derived mast cells. Biol Pharm Bull. 2004;27:786-788.
  17. Wang YR, Xu Y, Jiang ZZ, Guerram M, Wang B, Zhu X, Zhang LY. Deoxypodophyllotoxin Induces G2/M cell cycle arrest and apoptosis in SGC-7901 cells and inhibits tumor growth in vivo. Molecules. 2015;20:1661-1675. doi:10.3390/molecules20011661.
  18. Jung CH, Kim H, Ahn J, Jung SK, Um MY, Son KH, Kim TW, Ha TY. Anthricin Isolated from Anthriscus sylvestris (L.) Hoffm. inhibits the growth of Breast cancer cells by inhibiting Akt/mTOR signaling, and its apoptosis effects are enhanced by autophagy inhibition. Evid Based Complement Alternat Med. 2013;2013:9. doi:10.1155/2013/385219.
  19. Sang CY, Xu XH, Qin WW, Liu JF, Huui L, Chen SW. DPMA, a deoxypodophyllotoxin derivative, induces apoptosis and anti-angiogenesis in non-small cell lung cancer A549 cells. Bioorg Med Chem Lett. 2013;15:6650-6655. doi:10.1016/j.bmcl.2013.10.048.