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HepG2 인체 간암세포의 ROS 생성 및 ERK/Akt 신호전달 경로 조절을 통한 sanguinarine의 apoptosis 유도

황주영;최영현
Hwang, Ju Yeong;Cho, Yung Hyun

  • 투고 : 2015.07.13
  • 심사 : 2015.09.07
  • 발행 : 2015.09.30

초록

혈근초(Sanguinaria canadensis)에서 처음 분리된 sanguinarine은 항산화, 항암 및 면역 증강 등의 효능이 있는 것으로 알려진 alkaloid 계열 물질 중의 하나이다. 본 연구에서는 인체간암 HepG2 세포를 대상으로 sanguinarine의 apoptosis 유도 효능 및 관련 기전 해석을 시도하였다. 본 연구의 결과에 의하면 sanguinarine은 HepG2 간암세포의 증식을 처리 농도 의존적으로 억제하였으며, 이는 apoptosis 유도와 연관성이 있었다. Sanguinarine에 의한 apoptosis 유도에는 Fas 및 Bax의 발현 증가, 미토콘드리아에서 세포질로의 cytochrome c 유리 및 MMPl (Δψm)의 소실을 동반하였다. Sanguinarine은 intrinsic 및 extrinsic apoptosis pathway의 활성에 관여하는 initiator caspase인 caspase-9와 -8의 활성과 effector caspase인 caspase-3의 활성 및 PARP 단백질의 단편화를 유발하였다. Sanguinarine은 또한 ROS의 생성을 촉진시켰으며, N-acetylcysteine 처리에 의한 ROS의 생성을 차단하였을 경우, sanguinarine에 의한 apoptosis 효능이 완벽하게 차단되었다. 아울러 sanguinarine은 Akt의 인산화를 억제한 반면, MAPKs의 인산화를 촉진시켰으며, 특히 PI3K와 ERK의 선택적 억제제는 sanguinarine에 의한 HepG2 간암세포의 증식을 더욱 억제시켰다. 따라서 sanguinarine에 의한 HepG2 간암세포의 apoptosis 유발에는 ROS 생성 의존적인 intrinsic 및 extrinsic signaling pathway가 동시에 활성화되며, PI3K/Akt 및 ERK 신호계가 관여함을 알 수 있었다.

키워드

Apoptosis;extracellular signal-regulated kinase (ERK);PI3K/Akt;reactive oxygen species (ROS);sanguinarine

참고문헌

  1. Xu, J. Y., Meng, Q. H., Chong, Y., Jiao, Y., Zhao, L., Rosen, E. M. and Fan, S. 2013. Sanguinarine is a novel VEGF inhibitor involved in the suppression of angiogenesis and cell migration. Mol. Clin. Oncol. 1, 331-336.
  2. Yang, Y., Zhu, X., Chen, Y., Wang, X. and Chen, R. 2007. p38 and JNK MAPK, but not ERK1/2 MAPK, play important role in colchicine-induced cortical neurons apoptosis. Eur. J. Pharmacol. 576, 26-33. https://doi.org/10.1016/j.ejphar.2007.07.067
  3. Pica, F., Balestrieri, E., Serafino, A., Sorrentino, R., Gaziano, R., Moroni, G., Moroni, N., Palmieri, G., Mattei, M., Garaci, E. and Sinibaldi-Vallebona, P. 2012. Antitumor effects of the benzophenanthridine alkaloid sanguinarine in a rat syngeneic model of colorectal cancer. Anticancer Drugs 23, 32-42. https://doi.org/10.1097/CAD.0b013e32834a0c8e
  4. Slunská, Z., Gelnarová, E., Hammerová, J., Táborská, E. and Slaninová, I. 2010. Effect of quaternary benzo[c]phenanthridine alkaloids sanguilutine and chelilutine on normal and cancer cells. Toxicol. In Vitro 24, 697-706. https://doi.org/10.1016/j.tiv.2010.01.012
  5. Sun, M., Liu, C., Nadiminty, N., Lou, W., Zhu, Y., Yang, J., Evans, C. P., Zhou, Q. and Gao, A. C. 2012. Inhibition of Stat3 activation by sanguinarine suppresses prostate cancer cell growth and invasion. Prostate 72, 82-89. https://doi.org/10.1002/pros.21409
  6. Toker, A. and Yoeli-Lerner, M. 2006. Akt signaling and cancer: surviving but not moving on. Cancer Res. 66, 3963-3966. https://doi.org/10.1158/0008-5472.CAN-06-0743
  7. Vlachojannis, C., Magora, F. and Chrubasik, S. 2012. Rise and fall of oral health products with Canadian bloodroot extract. Phytother. Res. 26, 1423-1426.
  8. Vogt, A., Tamewitz, A., Skoko, J., Sikorski, R. P., Giuliano, K. A. and Lazo, J. S. 2005. The benzo[c]phenanthridine alkaloid, sanguinarine, is a selective, cell-active inhibitor of mitogen-activated protein kinase phosphatase-1. J. Biol. Chem. 280, 19078-19086. https://doi.org/10.1074/jbc.M501467200
  9. Weerasinghe, P., Hallock, S., Tang, S. C., Trump, B. and Liepins, A. 2006. Sanguinarine overcomes P-glycoprotein-mediatedmultidrug-resistance via induction of apoptosis and oncosis in CEM-VLB 1000 cells. Exp. Toxicol. Pathol. 58, 21-30. https://doi.org/10.1016/j.etp.2006.01.008
  10. Xu, J. Y., Meng, Q. H., Chong, Y., Jiao, Y., Zhao, L., Rosen, E. M. and Fan, S. 2012. Sanguinarine inhibits growth of human cervical cancer cells through the induction of apoptosis. Oncol. Rep. 28, 2264-2270.
  11. Jourdain, A. and Martinou, J. C. 2009. Mitochondrial outer-membrane permeabilization and remodelling in apoptosis. Int. J. Biochem. Cell Biol. 41, 1884-1889. https://doi.org/10.1016/j.biocel.2009.05.001
  12. Kadenbach, B., Arnold, S., Lee, I. and Hüttemann, M. 2004. The possible role of cytochrome c oxidase in stress-induced apoptosis and degenerative diseases. Biochim. Biophys. Acta. 1655, 400-408. https://doi.org/10.1016/j.bbabio.2003.06.005
  13. Kim, S., Lee, T. J., Leem, J., Choi, K. S., Park, J. W. and Kwon, T. K. 2008. Sanguinarine-induced apoptosis: generation of ROS, down-regulation of Bcl-2, c-FLIP, and synergy with TRAIL. J. Cell. Biochem. 104, 895-907. https://doi.org/10.1002/jcb.21672
  14. Laster, L. L. and Lobene, R. R. 1990. New perspectives on Sanguinaria clinicals: individual toothpaste and oral rinse testing. J. Can. Dent. Assoc. 56, 19-30.
  15. Lee, J. S., Jung, W. K., Jeong, M. H., Yoon, T. R. and Kim, H. K. 2012. Sanguinarine induces apoptosis of HT-29 human colon cancer cells via the regulation of Bax/Bcl-2 ratio and caspase-9-dependent pathway. Int. J. Toxicol. 31, 70-77. https://doi.org/10.1177/1091581811423845
  16. Mackraj, I., Govender, T. and Gathiram, P. 2008. Sanguinarine. Cardiovasc. Ther. 26, 75-83.
  17. Park, H., Bergeron, E., Senta, H., Guillemette, K., Beauvais, S., Blouin, R., Sirois, J. and Faucheux, N. 2010. Sanguinarine induces apoptosis of human osteosarcoma cells through the extrinsic and intrinsic pathways. Biochem. Biophys. Res. Commun. 399, 446-451. https://doi.org/10.1016/j.bbrc.2010.07.114
  18. Park, S. Y., Jin, M. L., Kim, Y. H., Lee, S. J. and Park, G. 2014. Sanguinarine inhibits invasiveness and the MMP-9 and COX-2 expression in TPA-induced breast cancer cells by inducing HO-1 expression. Oncol. Rep. 31, 497-504.
  19. Gupta, S. C., Kim, J. H., Prasad, S. and Aggarwal, B. B. 2010. Regulation of survival, proliferation, invasion, angiogenesis, and metastasis of tumor cells through modulation of inflammatory pathways by nutraceuticals. Cancer Metastasis Rev. 29, 405-434. https://doi.org/10.1007/s10555-010-9235-2
  20. Han, M. H., Kim, S. O., Kim, G. Y., Kwon, T. K., Choi, B. T., Lee, W. H. and Choi, Y. H. 2007. Induction of apoptosis by sanguinarine in C6 rat glioblastoma cells is associated with the modulation of the Bcl-2 family and activation of caspases through downregulation of extracellular signal-regulated kinase and Akt. Anticancer Drugs 18, 913-921.
  21. Han, M. H., Yoo, Y. H. and Choi, Y. H. 2008. Sanguinarine-induced apoptosis in human leukemia U937 cells via Bcl-2 downregulation and caspase-3 activation. Chemotherapy 54,157-165. https://doi.org/10.1159/000140359
  22. Han, M. H., Park, C., Jin, C. Y., Kim, G. Y., Chang, Y. C., Moon, S. K., Kim, W. J. and Choi, Y. H. 2013. Apoptosis induction of human bladder cancer cells by sanguinarine through reactive oxygen species-mediated up-regulation of early growth response gene-1. PLoS One 8, e63425. https://doi.org/10.1371/journal.pone.0063425
  23. Han, M. H., Kim, G. Y., Yoo, Y. H. and Choi, Y. H. 2013. Sanguinarine induces apoptosis in human colorectal cancer HCT-116 cells through ROS-mediated Egr-1 activation and mitochondrial dysfunction. Toxicol. Lett. 220, 157-166. https://doi.org/10.1016/j.toxlet.2013.04.020
  24. Hu, L., Hofmann, J., Lu, Y., Mills, G. B. and Jaffe, R. B. 2002. Inhibition of phosphatidylinositol 3'-kinase increases efficacy of paclitaxel in in vitro and in vivo ovarian cancer models. Cancer Res. 62, 1087-1092.
  25. Jeong, S. Y. and Seol, D. W. 2008. The role of mitochondria in apoptosis. BMB Rep. 41, 1122.
  26. Jin, Z. and El-Deiry, W. S. 2005. Overview of cell death signaling pathways. Cancer Biol. Ther. 4, 139-163.
  27. Degli Esposti, M. and Dive, C. 2003. Mitochondrial membrane permeabilisation by Bax/Bak. Biochem. Biophys. Res.Commun. 304, 455-461. https://doi.org/10.1016/S0006-291X(03)00617-X
  28. Dong, X. Z., Zhang, M., Wang, K., Liu, P., Guo, D. H., Zheng, X. L. and Ge, X. Y. 2013. Sanguinarine inhibits vascular endothelial growth factor release by generation of reactive oxygen species in MCF-7 human mammary adenocarcinoma cells. Biomed. Res. Int. 2013, 517698.
  29. Duriez, P. J. and Shah, G. M. 1997. Cleavage of poly(ADP-ribose) polymerase: a sensitive parameter to study cell death. Biochem. Cell. Biol. 75, 337-349. https://doi.org/10.1139/o97-043
  30. Duvoix, A., Delhalle, S., Blasius, R., Schnekenburger, M., Morceau, F., Fougère, M., Henry, E., Galteau, M. M., Dicato, M. and Diederich, M. 2004. Effect of chemopreventive agents on glutathione S-transferase P1-1 gene expression mechanisms via activating protein 1 and nuclear factor kappaB inhibition. Biochem. Pharmacol. 68, 1101-1111. https://doi.org/10.1016/j.bcp.2004.05.032
  31. Fulda, S. and Debatin, K. M. 2006. Extrinsic versus intrinsic apoptosis pathways in anticancer chemotherapy. Oncogene 25, 4798-4811. https://doi.org/10.1038/sj.onc.1209608
  32. Gatti, L., Cossa, G., Tinelli, S., Carenini, N., Arrighetti, N., Pennati, M., Cominetti, D., De Cesare, M., Zunino, F.,Zaffaroni, N. and Perego, P. 2014. Improved apoptotic cell death in drug-resistant non-small-cell lung cancer cells by tumor necrosis factor-related apoptosis-inducing ligandbased treatment. J. Pharmacol. Exp. Ther. 348, 360-371. https://doi.org/10.1124/jpet.113.210054
  33. Godowski, K. C. 1989. Antimicrobial action of sanguinarine. J. Clin. Dent. 1, 96-101.
  34. Gu, S., Yang, X. C., Xiang, X. Y., Wu, Y., Zhang, Y., Yan, X. Y., Xue, Y. N., Sun, L. K. and Shao, G. G. 2015. Sanguinarine-induced apoptosis in lung adenocarcinoma cells is dependent on reactive oxygen species production and endoplasmic reticulum stress. Oncol. Rep. 34, 913-919.
  35. Brenner, D. and Mak, T. W. 2009. Mitochondrial cell death effectors. Curr. Opin. Cell Biol. 21, 871-877. https://doi.org/10.1016/j.ceb.2009.09.004
  36. Choi, W. Y., Kim, G. Y., Lee, W. H. and Choi, Y. H. 2008. Sanguinarine, a benzophenanthridine alkaloid, induces apoptosis in MDA-MB-231 human breast carcinoma cells through a reactive oxygen species-mediated mitochondrial pathway. Chemotherapy 54, 279-287. https://doi.org/10.1159/000149719
  37. Choi, W. Y., Jin, C. Y., Han, M. H., Kim, G. Y., Kim, N. D., Lee, W. H., Kim, S. K. and Choi, Y. H. 2009. Sanguinarine sensitizes human gastric adenocarcinoma AGS cells to TRAIL-mediated apoptosis via down-regulation of AKT and activation of caspase-3. Anticancer Res. 29, 4457-4465.
  38. Choi, Y. H., Choi, W. Y., Hong, S. H., Kim, S. O., Kim, G. Y., Lee, W. H. and Yoo, Y. H. 2009. Anti-invasive activity of sanguinarine through modulation of tight junctions and matrix metalloproteinase activities in MDA-MB-231 human breast carcinoma cells. Chem. Biol. Interact. 179, 185-191. https://doi.org/10.1016/j.cbi.2008.11.009
  39. Circu, M. L. and Aw, T. Y. 2001. Reactive oxygen species, cellular redox systems, and apoptosis. Free. Radic. Biol. Med. 48, 749-762.
  40. De Stefano, I., Raspaglio, G., Zannoni, G. F., Travaglia, D., Prisco, M. G., Mosca, M., Ferlini, C., Scambia, G. and Gallo, D. 2009. Antiproliferative and antiangiogenic effects of the benzophenanthridine alkaloid sanguinarine in melanoma. Biochem. Pharmacol. 78, 1374-1381. https://doi.org/10.1016/j.bcp.2009.07.011
  41. Bassi, R., Heads, R., Marber, M. S. and Clark, J. E. 2008. Targeting p38-MAPK in the ischaemic heart: kill or cure? Curr. Opin. Pharmacol. 8, 141-146. https://doi.org/10.1016/j.coph.2008.01.002
  42. Bedner, E., Li, X., Gorczyca, W., Melamed, M. R. and Darzynkiewicz, Z. 1999. Analysis of apoptosis by laser scanning cytometry. Cytometry 35, 181-195. https://doi.org/10.1002/(SICI)1097-0320(19990301)35:3<181::AID-CYTO1>3.0.CO;2-5
  43. Adhami, V. M., Aziz, M. H., Mukhtar, H. and Ahmad, N. 2003. Activation of prodeath Bcl-2 family proteins and mitochondrial apoptosis pathway by sanguinarine in immortalized human HaCaT keratinocytes. Clin. Cancer Res. 9, 3176-3182.
  44. Ahsan, H., Reagan-Shaw, S., Breur, J. and Ahmad, N. 2007. Sanguinarine induces apoptosis of human pancreatic carcinoma AsPC-1 and BxPC-3 cells via modulations in Bcl-2 family proteins. Cancer Lett. 249, 198-208. https://doi.org/10.1016/j.canlet.2006.08.018
  45. Basini, G., Santini, S. E., Bussolati, S. and Grasselli, F. 2007. Sanguinarine inhibits VEGF-induced Akt phosphorylation. Ann. N. Y. Acad. Sci. 1095, 371-376. https://doi.org/10.1196/annals.1397.040

피인용 문헌

  1. Extract from Artemisia annua Linn? Induces Apoptosis through the Mitochondrial Signaling Pathway in HepG2 Cells vol.45, pp.12, 2016, https://doi.org/10.5352/JLS.2015.25.9.984