Naunyn-Schmiedeberg's Archives of Pharmacology

, Volume 386, Issue 11, pp 1009–1016 | Cite as

Phenethyl isothiocyanate induces apoptosis of cholangiocarcinoma cells through interruption of glutathione and mitochondrial pathway

  • Ornanong Tusskorn
  • Auemduan Prawan
  • Laddawan Senggunprai
  • Upa Kukongviriyapan
  • Veerapol Kukongviriyapan
Original Article


Phenethyl isothiocyanate (PEITC) is a natural isothiocyanate with anticancer activity against many drug-resistant cancer cells. A body of evidence suggests that PEITC enhances oxidative stress leading to cancer cell death. Cholangiocarcinoma (CCA) is an aggressive bile duct cancer with resistance to chemotherapeutic drugs. PEITC rapidly kills KKU-100 CCA cells with concurrent induction of cellular glutathione depletion, superoxide formation, and loss of mitochondrial transmembrane potential. The loss was associated with increased Bax and decreased Bcl-xl proteins followed by the release of cytochrome c and the activation of caspase-9 and -3. Although TEMPOL could prevent superoxide formation, it did not prevent the disruption of glutathione (GSH) redox, mitochondrial dysfunction, and cell death. On the other hand, N-acetylcysteine could prevent the events and cell death. It was concluded that disruption of GSH redox but not superoxide formation may be an initial step leading to mitochondrial injury. PEITC could be a promising chemopreventive agent for CCA.


Phenethyl isothiocyanate Cancer chemoprevention Cholangiocarcinoma Mitochondrial dysfunction Glutathione 



Buthionine sulfoximine






Phenethyl isothiocyanate




Glutathione disulfide


Mitochondrial transmembrane potential




Reactive oxygen species


Sulforhodamine B


5,5′,6,′-Tetrachloro-1,1′,3,3′-tetraethylbenzimidazolyl-carbocyanine iodide


Mitochondrial outer membrane permeabilization


  1. Buranrat B, Prawan A, Kukongviriyapan U, Kongpetch S, Kukongviriyapan V (2010) Dicoumarol enhances gemcitabine-induced cytotoxicity in high NQO1-expressing cholangiocarcinoma cells. World J Gastroenterol 16:2362–2370PubMedCrossRefGoogle Scholar
  2. Chan JM, Wang F, Holly EA (2005) Vegetable and fruit intake and pancreatic cancer in a population-based case–control study in the San Francisco bay area. Cancer Epidemiol Biomarkers Prev 14:2093–2097PubMedCrossRefGoogle Scholar
  3. Cheung KL, Khor TO, Yu S, Kong AN (2008) PEITC induces G1 cell cycle arrest on HT-29 cells through the activation of p38 MAPK signaling pathway. AAPS J 10:277–281PubMedCrossRefGoogle Scholar
  4. Cheung KL, Kong AN (2010) Molecular targets of dietary phenethyl isothiocyanate and sulforaphane for cancer chemoprevention. AAPS J 12:87–97PubMedCrossRefGoogle Scholar
  5. Chiao JW, Wu H, Ramaswamy G, Conaway CC, Chung FL, Wang L, Liu D (2004) Ingestion of an isothiocyanate metabolite from cruciferous vegetables inhibits growth of human prostate cancer cell xenografts by apoptosis and cell cycle arrest. Carcinogenesis 25:1403–1408PubMedCrossRefGoogle Scholar
  6. Chipuk JE, Green DR (2008) How do BCL-2 proteins induce mitochondrial outer membrane permeabilization? Trends Cell Biol 18:157–164PubMedCrossRefGoogle Scholar
  7. Eckel F, Schmid RM (2007) Chemotherapy in advanced biliary tract carcinoma: a pooled analysis of clinical trials. Br J Cancer 96:896–902PubMedCrossRefGoogle Scholar
  8. Hayes JD, Kelleher MO, Eggleston IM (2008) The cancer chemopreventive actions of phytochemicals derived from glucosinolates. Eur J Nutr 47(Suppl 2):73–88PubMedCrossRefGoogle Scholar
  9. Hezel AF, Zhu AX (2008) Systemic therapy for biliary tract cancers. Oncologist 13:415–423PubMedCrossRefGoogle Scholar
  10. Keum YS, Owuor ED, Kim BR, Hu R, Kong AN (2003) Involvement of Nrf2 and JNK1 in the activation of antioxidant responsive element (ARE) by chemopreventive agent phenethyl isothiocyanate (PEITC). Pharm Res 20:1351–1356PubMedCrossRefGoogle Scholar
  11. Khan SA, Thomas HC, Davidson BR, Taylor-Robinson SD (2005) Cholangiocarcinoma. Lancet 366:1303–1314PubMedCrossRefGoogle Scholar
  12. Khor TO, Cheung WK, Prawan A, Reddy BS, Kong AN (2008) Chemoprevention of familial adenomatous polyposis in Apc(Min/+) mice by phenethyl isothiocyanate (PEITC). Mol Carcinog 47:321–325PubMedCrossRefGoogle Scholar
  13. Kim JS, He L, Lemasters JJ (2003) Mitochondrial permeability transition: a common pathway to necrosis and apoptosis. Biochem Biophys Res Commun 304:463–470PubMedCrossRefGoogle Scholar
  14. Kluza J, Jendoubi M, Ballot C, Dammak A, Jonneaux A, Idziorek T, Joha S, Dauphin V, Malet-Martino M, Balayssac S, Maboudou P, Briand G, Formstecher P, Quesnel B, Marchetti P (2011) Exploiting mitochondrial dysfunction for effective elimination of imatinib-resistant leukemic cells. PLoS One 6:e21924PubMedCrossRefGoogle Scholar
  15. Kongpetch S, Kukongviriyapan V, Prawan A, Senggunprai L, Kukongviriyapan U, Buranrat B (2012) Crucial role of heme oxygenase-1 on the sensitivity of cholangiocarcinoma cells to chemotherapeutic agents. PLoS One 7:e34994PubMedCrossRefGoogle Scholar
  16. Krishna MC, Russo A, Mitchell JB, Goldstein S, Dafni H, Samuni A (1996) Do nitroxide antioxidants act as scavengers of O2-. or as SOD mimics? J Biol Chem 271:26026–26031PubMedCrossRefGoogle Scholar
  17. Lee S, Kim WH, Jung HY, Yang MH, Kang GH (2002) Aberrant CpG island methylation of multiple genes in intrahepatic cholangiocarcinoma. Am J Pathol 161:1015–1022PubMedCrossRefGoogle Scholar
  18. Lee Y, Kim YJ, Choi YJ, Lee JW, Lee S, Chung HW (2012) Enhancement of cisplatin cytotoxicity by benzyl isothiocyanate in HL-60 cells. Food Chem Toxicol 50:2397–2406PubMedCrossRefGoogle Scholar
  19. Mi L, Gan N, Chung FL (2011a) Isothiocyanates inhibit proteasome activity and proliferation of multiple myeloma cells. Carcinogenesis 32:216–223PubMedCrossRefGoogle Scholar
  20. Mi L, Hood BL, Stewart NA, Xiao Z, Govind S, Wang X, Conrads TP, Veenstra TD, Chung FL (2011b) Identification of potential protein targets of isothiocyanates by proteomics. Chem Res Toxicol 24:1735–1743PubMedCrossRefGoogle Scholar
  21. Mi L, Wang X, Govind S, Hood BL, Veenstra TD, Conrads TP, Saha DT, Goldman R, Chung FL (2007) The role of protein binding in induction of apoptosis by phenethyl isothiocyanate and sulforaphane in human non-small lung cancer cells. Cancer Res 67:6409–6416PubMedCrossRefGoogle Scholar
  22. Mukherjee S, Bhattacharya RK, Roy M (2009) Targeting protein kinase C (PKC) and telomerase by phenethyl isothiocyanate (PEITC) sensitizes PC-3 cells towards chemotherapeutic drug-induced apoptosis. J Environ Pathol Toxicol Oncol 28:269–282PubMedCrossRefGoogle Scholar
  23. Norberg E, Gogvadze V, Vakifahmetoglu H, Orrenius S, Zhivotovsky B (2010) Oxidative modification sensitizes mitochondrial apoptosis-inducing factor to calpain-mediated processing. Free Radic Biol Med 48:791–797PubMedCrossRefGoogle Scholar
  24. Parasassi T, Brunelli R, Costa G, De Spirito M, Krasnowska E, Lundeberg T, Pittaluga E, Ursini F (2010) Thiol redox transitions in cell signaling: a lesson from N-acetylcysteine. ScientificWorldJournal 10:1192–1202PubMedCrossRefGoogle Scholar
  25. Patel T (2011) Cholangiocarcinoma—controversies and challenges. Nat Rev Gastroenterol Hepatol 8:189–200PubMedCrossRefGoogle Scholar
  26. Rose P, Whiteman M, Huang SH, Halliwell B, Ong CN (2003) beta-Phenylethyl isothiocyanate-mediated apoptosis in hepatoma HepG2 cells. Cell Mol Life Sci 60:1489–1503PubMedCrossRefGoogle Scholar
  27. Sripa B, Brindley PJ, Mulvenna J, Laha T, Smout MJ, Mairiang E, Bethony JM, Loukas A (2012) The tumorigenic liver fluke Opisthorchis viverrini—multiple pathways to cancer. Trends Parasitol 28:395–407PubMedCrossRefGoogle Scholar
  28. Sripa B, Leungwattanawanit S, Nitta T, Wongkham C, Bhudhisawasdi V, Puapairoj A, Sripa C, Miwa M (2005) Establishment and characterization of an opisthorchiasis-associated cholangiocarcinoma cell line (KKU-100). World J Gastroenterol 11:3392–3397PubMedGoogle Scholar
  29. Suphim B, Prawan A, Kukongviriyapan U, Kongpetch S, Buranrat B, Kukongviriyapan V (2010) Redox modulation and human bile duct cancer inhibition by curcumin. Food Chem Toxicol 48:2265–2272PubMedCrossRefGoogle Scholar
  30. Trachootham D, Zhang H, Zhang W, Feng L, Du M, Zhou Y, Chen Z, Pelicano H, Plunkett W, Wierda WG, Keating MJ, Huang P (2008) Effective elimination of fludarabine-resistant CLL cells by PEITC through a redox-mediated mechanism. Blood 112:1912–1922PubMedCrossRefGoogle Scholar
  31. Xiao D, Powolny AA, Moura MB, Kelley EE, Bommareddy A, Kim SH, Hahm ER, Normolle D, Van Houten B, Singh SV (2010) Phenethyl isothiocyanate inhibits oxidative phosphorylation to trigger reactive oxygen species-mediated death of human prostate cancer cells. J Biol Chem 285:26558–26569PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Ornanong Tusskorn
    • 1
  • Auemduan Prawan
    • 1
    • 2
  • Laddawan Senggunprai
    • 1
    • 2
  • Upa Kukongviriyapan
    • 3
  • Veerapol Kukongviriyapan
    • 1
    • 2
  1. 1.Department of Pharmacology, Faculty of MedicineKhon Kaen UniversityKhon KaenThailand
  2. 2.Liver Fluke and Cholangiocarcinoma Research CenterKhon Kaen UniversityKhon KaenThailand
  3. 3.Department of Physiology, Faculty of MedicineKhon Kaen UniversityKhon KaenThailand

Personalised recommendations