The Journal of Korean Medicine (대한한의학회지)

The Society of Korean Medicine (대한한의학회)
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Orostachys japonicus DW and EtOH Extracts Induce Apoptosis in Cholangiocarcinoma Cell Line SNU-1079

  • Choi, Eun Sol (Department of Clinical Korean Medicine, Graduate School, Kyung Hee University) ;
  • Lee, Jang Hoon (Department of Clinical Korean Medicine, Graduate School, Kyung Hee University)
  • Received : 2015.11.16
  • Accepted : 2015.12.24
  • Published : 2015.12.30
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Abstract

Objectives: This study was performed to investigate the anti-tumor effect of O. japonicus extracts on intrahepatic cholangiocarcinoma cell line SNU-1079. Methods: Cholangiocarcinoma SNU-1079 cells were treated with various concentrations of O. japonicus DW and EtOH extracts ($0-300{\mu}g/ml$) for 24, 48 or 72 h. Cell viability was evaluated through a PMS/MTS assay, and the apoptosis rate was examined through ELISA assay and flow cytometry analysis. The mRNA expression of apoptosis- and cell cycle progression-related genes (Bcl-2, Mcl-1, Bax, Survivin, Cyclin D1, and p21) was evaluated using real-time PCR, and the caspase activity was examined using immunoblot analysis. Results: O. japonicus extracts inhibited cell proliferation and increased apoptosis rate in both ELISA assay and flow cytometry analysis. O. japonicus extracts decreased Bcl-2, Mcl-1, Survivin, and Cyclin D1 mRNA expression and increased Bax mRNA level. O. japonicus extracts also increased Caspase-3 activation. Overall, O. japonicus DW extracts were more effective than EtOH extracts. Conclusions: O. japonicus inhibited cell proliferation and induced apoptosis in SNU-1079 cells via mitochondria -mediated intrinsic pathway, which leads to Caspase-3 activation. The results indicate that O. japonicus is a potential therapeutic herb with anti-tumor effect against intrahepatic cholangiocarcinoma.

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References

  1. Rizvi S, Gores GJ. Pathogenesis, diagnosis, and management of cholangiocarcinoma. Gastroenterology. 2013;145:1215-29. https://doi.org/10.1053/j.gastro.2013.10.013
  2. Shaib Y, El-Serag H.B. The epidemiology of cholangiocarcinoma. Seminars in liver disease 2004(24)115-25.
  3. Welzel TM, McGlynn KA, Hsing AW, O'Brien TR., Pfeiffer RM. Impact of classification of hilar cholangiocarcinomas (Klatskin tumors) on the incidence of intra- and extrahepatic cholangiocarcinoma in the United States. Journal of the National Cancer Institute. 2006;98:873-5. https://doi.org/10.1093/jnci/djj234
  4. Fava G, Lorenzini I. Molecular pathogenesis of cholangiocarcinoma. Int J Hepatol. 2012;2012:1-7.
  5. Lazaridis KN, Gores GJ. Cholangiocarcinoma. Gastroenterology. 2005;128:1655-67. https://doi.org/10.1053/j.gastro.2005.03.040
  6. Sripa B, Pairojkul C. Cholangiocarcinoma: lessons from Thailand. Current opinion in gastroenterology. 2008;24:349-56. https://doi.org/10.1097/MOG.0b013e3282fbf9b3
  7. Khan SA, Taylor-Robinson SD, Toledano M.B, Beck A, Elliott P, Thomas HC. Changing international trends in mortality rates for liver, biliary and pancreatic tumours. Journal of hepatology. 2002;37:806-13. https://doi.org/10.1016/S0168-8278(02)00297-0
  8. Khan SA., Toledano MB, Taylor-Robinson SD. Epidemiology, risk factors, and pathogenesis of cholangiocarcinoma. HPB : the official journal of the International Hepato Pancreato Biliary Association. 2008;10:77-82. https://doi.org/10.1080/13651820801992641
  9. McGlynn KA, Tarone RE, El-Serag HB. A comparison of trends in the incidence of hepatocellular carcinoma and intrahepatic cholangiocarcinoma in the United States. Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology. 2006;15:1198-203. https://doi.org/10.1158/1055-9965.EPI-05-0811
  10. Patel, T. Increasing incidence and mortality of primary intrahepatic cholangiocarcinoma in the United States. Hepatology. 2001;33:1353-7. https://doi.org/10.1053/jhep.2001.25087
  11. Fava G, Marzioni M, Benedetti A, Glaser S, DeMorrow S, Francis H, et al. Molecular pathology of biliary tract cancers. Cancer letters. 2007;250:155-67. https://doi.org/10.1016/j.canlet.2006.09.011
  12. Sandhu DS, Shire AM, Roberts LR. Epigenetic DNA hypermethylation in cholangiocarcinoma: potential roles in pathogenesis, diagnosis and identification of treatment targets. Liver international : official journal of the International Association for the Study of the Liver. 2008;28:12-27.
  13. Tischoff I, Wittekind C, Tannapfel A. Role of epigenetic alterations in cholangiocarcinoma. Journal of hepato-biliary-pancreatic surgery. 2006;13:274-9. https://doi.org/10.1007/s00534-005-1055-3
  14. Lowe SW, Lin AW. Apoptosis in cancer. Carcinogenesis. 2000;21:485-95. https://doi.org/10.1093/carcin/21.3.485
  15. Thompson CB, Apoptosis in the pathogenesis and treatment of disease. Science. 1995;267:1456-62. https://doi.org/10.1126/science.7878464
  16. Danial NN, Korsmeyer SJ. Cell death: critical control points. Cell. 2004;116:205-19. https://doi.org/10.1016/S0092-8674(04)00046-7
  17. Taylor RC, Cullen SP, Martin SJ. Apoptosis: controlled demolition at the cellular level. Nature reviews. Molecular cell biology. 2008;9: 231-41.
  18. Tsuruo T, Naito M, Tomida A, Fujita, N, Mashima T, Sakamoto H, et al. Molecular targeting therapy of cancer: drug resistance, apoptosis and survival signal. Cancer science. 2003;94:15-21. https://doi.org/10.1111/j.1349-7006.2003.tb01345.x
  19. Debatin KM, Apoptosis pathways in cancer and cancer therapy. Cancer immunology, immunotherapy:CII. 2004;53:153-9. https://doi.org/10.1007/s00262-003-0474-8
  20. Lee HS, Ryu DS, Lee GS, Lee DS. Anti-inflammatory effects of dichloromethane fraction from Orostachys japonicus in RAW 264.7 cells: suppression of NF-kappaB activation and MAPK signaling. Journal of ethnopharmacology. 2012;140:271-6. https://doi.org/10.1016/j.jep.2012.01.016
  21. Yoon Y, Kim KS, Hong SG, Kang BJ, Lee M Y, Cho DW. Protective effects of Orostachys japonicus A. Berger (Crassulaceae) on H2O2-induced apoptosis in GT1-1 mouse hypothalamic neuronal cell line. Journal of ethnopharmacology. 2000;69:73-8. https://doi.org/10.1016/S0378-8741(99)00107-5
  22. Jung HJ, Choi J, Nam JH, Park HJ. Anti-ulcerogenic effects of the flavonoid-rich fraction from the extract of Orostachys japonicus in mice. Journal of medicinal food. 2007;10:702-6. https://doi.org/10.1089/jmf.2006.223
  23. Park JC, Han WD, Park JR, Choi SH, Choi J W. Changes in hepatic drug metabolizing enzymes and lipid peroxidation by methanol extract and major compound of Orostachys japonicus. Journal of ethnopharmacology. 2005;102:313-8. https://doi.org/10.1016/j.jep.2005.06.023
  24. Shin DY, Lee WS, Jung JH, Hong SH, Park C, Kim HJ, et al. Flavonoids from Orostachys japonicus A. Berger inhibit the invasion of LnCaP prostate carcinoma cells by inactivating Akt and modulating tight junctions. International journal of molecular sciences. 2013;14:18407-20. https://doi.org/10.3390/ijms140918407
  25. Ryu DS, Kim SH, Kwon JH, Lee DS. Orostachys japonicus induces apoptosis and cell cycle arrest through the mitochondria -dependent apoptotic pathway in AGS human gastric cancer cells. International journal of oncology. 2014;45:459-69. https://doi.org/10.3892/ijo.2014.2404
  26. Lee WS, Yun JW, Nagappan A, Jung JH, Yi SM, Kim DH et al. Flavonoids from Orostachys japonicus A. Berger induces caspase-dependent apoptosis at least partly through activation of p38 MAPK pathway in U937 human leukemic cells. Asian Pacific journal of cancer prevention : APJCP. 2015; 16:465-9. https://doi.org/10.7314/APJCP.2015.16.2.465
  27. Kim YI, Park SW, Choi IH, Lee JH, Woo HJ, Kim, Y. Effect of Orostachys japonicus on cell growth and apoptosis in human hepatic stellate cell line LX2. The American journal of Chinese medicine. 2011;39:601-13. https://doi.org/10.1142/S0192415X11009068
  28. Yun KS, Lee JH, Woo Hj. Effect of Orostachys japonicus A. berger in Apoptosis in K562 Cell Lines. Korean J. Orient. Int. Med. 2006;27:166-177.
  29. Oh CH, Kim NS, Jeon HJ, Han KS, Lee MJ Lee, Kwon J. Effect of Orostachys japonicus A. Berger on Apoptosis Induction of Human Leukemia HL60 Cells. Kor. J. Pharmacogn. 2009;40:118-122.
  30. Kim JY, Won YS, Lee JH, Shin DY, Seo KI. Cultivated Orostachys japonicus Induces Apoptosis in Human Colon Cancer Cells. KOREAN J. FOOD SCI. TECHNOL. 2012;44:317-323. https://doi.org/10.9721/KJFST.2012.44.3.317
  31. Won YS, Kwon SJ, Ahn DU, Shin DY, Seo KI. Anticancer Effects of Cultivated Orostachys japonicus on Human Prostate Cancer Cells. J Korean Soc Food Sci Nutr. 2014;43:67-73. https://doi.org/10.3746/jkfn.2014.43.1.067
  32. Ku JL, Yoon KA, Kim IJ, Kim WH, Jang JY, Suh KS, et al. Establishment and characterisation of six human biliary tract cancer cell lines. British journal of cancer. 2002;87:187-93. https://doi.org/10.1038/sj.bjc.6600440
  33. Joung B, Kim Y. Study on Anti-Cancer Effects of Rhus Verniciflua Stokes Extracted with Sterile Distilled Water on Two Cholangiocarcinoma Cell Lines, SNU-1079 and SNU-1196. J. Int. Korean Med. 2015;36:1-12.
  34. Park JK. Pro-apoptotic Effects of Sanguisorbae Radix Ethanol-Extracts on Two Cholangiocarcinoma Cell Lines, SNU-1079 and SNU-1196. Korean J. Orient. Int. Med. 2012;33:465-475.
  35. Khan SA, Davidson BR, Goldin RD, Heaton N, Karani J, Pereira SP. Guidelines for the diagnosis and treatment of cholangiocarcinoma: an update. Gut. 2012;61:1657-69. https://doi.org/10.1136/gutjnl-2011-301748
  36. Tyson GL, El-Serag HB, Risk factors for cholangiocarcinoma. Hepatology. 2011;54:173-84.
  37. Shaib YH, Davila JA., McGlynn K, El-Serag HB. Rising incidence of intrahepatic cholangiocarcinoma in the United States: a true increase? Journal of hepatology. 2004;40:472-7. https://doi.org/10.1016/j.jhep.2003.11.030
  38. Alpini G, McGill JM, Larusso NF. The pathobiology of biliary epithelia. Hepatology. 2002;35:1256-68. https://doi.org/10.1053/jhep.2002.33541
  39. Harnois DM, Que FG, Celli A, LaRusso NF, Gores GJ. Bcl-2 is overexpressed and alters the threshold for apoptosis in a cholangiocarcinoma cell line. Hepatology. 1997;26:884-90. https://doi.org/10.1002/hep.510260413
  40. Taniai M, Grambihler A, Higuchi H, Werneburg N, Bronk SF, Farrugia DJ. et al. Mcl-1 mediates tumor necrosis factor-related apoptosis-inducing ligand resistance in human cholangiocarcinoma cells. Cancer research. 2004;64:3517-24. https://doi.org/10.1158/0008-5472.CAN-03-2770
  41. Rizvi S, Gores GJ. Molecular pathogenesis of cholangiocarcinoma. Digestive diseases. 2014;32:564-9. https://doi.org/10.1159/000360502
  42. Kobayashi S, Werneburg NW, Bronk SF, Kaufmann SH, Gores GJ. Interleukin-6 contributes to Mcl-1 up-regulation and TRAIL resistance via an Akt-signaling pathway in cholangiocarcinoma cells. Gastroenterology. 2005;128:2054-65. https://doi.org/10.1053/j.gastro.2005.03.010
  43. Shi Y, Chen J, Weng C, Chen R, Zheng Y, Chen Q, et al. Identification of the protein-protein contact site and interaction mode of human VDAC1 with Bcl-2 family proteins. Biochemical and biophysical research communications. 2003;305:989-96. https://doi.org/10.1016/S0006-291X(03)00871-4
  44. Kroemer G, Galluzzi L, Brenner C. Mitochondrial membrane permeabilization in cell death. Physiological reviews. 2007;87: 99-163. https://doi.org/10.1152/physrev.00013.2006
  45. Sah NK, Khan Z, Khan GJ, Bisen PS. Structural, functional and therapeutic biology of survivin. Cancer letters. 2006;244:164-71. https://doi.org/10.1016/j.canlet.2006.03.007
  46. Baldin V, Lukas J, Marcote MJ, Pagano M, Draetta G. Cyclin D1 is a nuclear protein required for cell cycle progression in G1. Genes & development. 1993;7:812-21. https://doi.org/10.1101/gad.7.5.812
  47. Diehl JA. Cycling to cancer with cyclin D1. Cancer biology & therapy. 2002;1:226-31. https://doi.org/10.4161/cbt.72
  48. Shintani M, Okazaki A, Masuda T, Kawada M, Ishizuka M, Doki Y, et al. Overexpression of cyclin DI contributes to malignant properties of esophageal tumor cells by increasing VEGF production and decreasing Fas expression. Anticancer research. 2002;22:639-47.
  49. Gartel AL, Radhakrishnan SK. Lost in transcription: p21 repression, mechanisms, and consequences. Cancer research. 2005;65:3980-5. https://doi.org/10.1158/0008-5472.CAN-04-3995
  50. Ghavami S, Hashemi M, Ande SR, Yeganeh B, Xiao W, Eshraghi M, et al. Apoptosis and cancer: mutations within caspase genes. Journal of medical genetics. 2009;46:497-510. https://doi.org/10.1136/jmg.2009.066944
  51. Porter AG, Janicke RU. Emerging roles of caspase-3 in apoptosis. Cell death and differentiation. 1999;6:99-104. https://doi.org/10.1038/sj.cdd.4400476

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