Andrographolide Inhibits Human Hepatoma-Derived Hep3B Cell Growth through the Activation of c-Jun N-Terminal Kinase

 

 Buy Article Permissions and Reprints

Abstract

Andrographolide (Andro) is a potentially anti-inflammatory diterpenoid lactone isolated from the traditional herbal medicine Andrographis paniculata, which has been effectively used for the treatment of infection, inflammation, cold, fever and diarrhea in China for centuries. In the current study, we found that Andro significantly decreased the number of surviving hepatoma-derived Hep3B cells in the MTT assay and induced cell apoptosis. Further study showed that Andro induced activation of mitogen-activated protein kinases (MAPKs) including p38 kinase, c-Jun N-terminal kinase (JNK) and extracellular signal-related kinases (ERK1/2), but had no significant effect on caspase-3, Bcl-xL and Bcl-2, which are apoptosis-related proteins. Moreover, inhibition of JNK activation partially rescued the toxic effect of Andro on Hep3B cells. Therefore, our results indicate that the JNK signaling pathway plays an important role in the toxic effect of Andro on Hep3B cells.

Abbreviations

Andro:andrographolide

CDK:cyclin-dependent kinase

ELISA:enzyme-linked immunosorbent assay

ERK1/2:extracellular signal-related kinases

FBS:fetal bovine serum

HCC:hepatocellular carcinoma

HPLC:high pressure liquid chromatography

JNK:c-Jun N-terminal kinase

MAPK:mitogen-activated protein kinase

MEM:modified Eagle's medium

RTKs:receptor tyrosine kinases

References

• 1 Chang H M, But P PH. Pharmacology and application of Chinese material medica. Vol. 1 Singapore; World Scientific Press 1987: 918-28.
  Google Scholar
• 2 Poolsup N, Suthisisang C, Prathanturarug S, Asawamekin A, Chanchareon U. Andrographis paniculata in the symptomatic treatment of uncomplicated upper respiratory tract infection: systematic review of randomized controlled trials.  J Clin Pharm Ther. 2004;  29 37-45.
  CrossrefPubMedGoogle Scholar
• 3 Xia Y F, Ye B Q, Li Y D, Wang J G, He X J, Lin X. et al . Andrographolide attenuates inflammation by inhibition of NF-kabba B activation through covalent modification of reduced cysteine 62 of p50.  J Immunol. 2004;  173 4207-17.
  PubMedGoogle Scholar
• 4 Jain D C, Gupta M M, Saxena S, Kumar S. LC analysis of hepatoprotective diterpenoids from Andrographis paniculata .  J Pharm Biomed Anal. 2000;  22 705-9.
  CrossrefPubMedGoogle Scholar
• 5 Kumar R A, Sridevi K, Kumar N V, Nanduri S, Rajagopal S. Anticancer and immunostimulatory compounds from Androgaphis paniculata .  J Ethnopharmacol. 2004;  92 291-5.
  CrossrefPubMedGoogle Scholar
• 6 Satyanarayana C, Deevi D S, Rajagopalan R, Srinivas N, Rajagopal S. DRF 3188 a novel semi-synthetic analog of andrographolide: cellular response to MCF 7 breast cancer cells.  BMC Cancer. 2004;  4 26.
  CrossrefPubMedGoogle Scholar
• 7 Rajagopal S, Kumar R A, Deevi D S, Satyanarayana C, Rajagopalan R. Andrographolide, a potential cancer therapeutic agent isolated from andrographis paniculata.  J Exp Ther Oncol. 2003;  3 147-58.
  CrossrefPubMedGoogle Scholar
• 8 Cheung H Y, Cheung S H, Li J, Cheung C S, Lai W P, Fong W F. et al . Andrographolide isolated from Andrographis paniculata induces cell cycle arrest and mitochondrial-mediated apoptosis in human leukemic HL-60 cells.  Planta Med. 2005;  71 1106-11.
  Article in Thieme ConnectPubMedGoogle Scholar
• 9 Zhou J, Zhang S Y, Ong C N, Shen H M. Critical role of pro-apoptotic Bcl-2 family members in andrographolide-induced apoptosis in human cancer cells.  Biochem Pharmacol. 2006;  72 132-44.
  CrossrefPubMedGoogle Scholar
• 10 Kim Y S, Milner J A. Targets for indole-3-carbinol in cancer prevention.  J Nutr Biochem. 2005;  16 65-73.
  PubMedGoogle Scholar
• 11 Jacobson M D, Weil M, Raff M C. Programmed cell death in the animal development.  Cell. 1997;  88 347-54.
  CrossrefPubMedGoogle Scholar
• 12 Natoni F, Diolordi L, Santoni C, Gilardini Montani M S. Sodium butyrate sensitizes human pancreatic cancer cells to both the intrinsic and the extrinsic apoptotic pathways.  Biochim Biophys Acta. 2005;  1745 318-29.
  PubMedGoogle Scholar
• 13 Cory S, Adams J M. The Bcl2 family: regulators of the cellular life-or-death switch.  Nat Rev Cancer. 2002;  2 647-56.
  CrossrefPubMedGoogle Scholar
• 14 Chang L, Karin M. Mammalian MAP kinase signalling cascades.  Nature. 2001;  410 37-40.
  CrossrefPubMedGoogle Scholar
• 15 Hunter T. Protein kinases and phosphatases: the yin and yang of protein phosphorylation and signaling.  Cell. 1995;  80 225-36.
  CrossrefPubMedGoogle Scholar
• 16 Galan A, Garcia-Bermejo M L, Troyano A, Vilaboa N E, de Blas E, Kazanietz M G. et al . Stimulation of p38 mitogen-activated protein kinase is an early regulatory event for the cadmium-induced apoptosis in human promonocytic cells.  J Biol Chem. 2000;  275 11 418-24.
  PubMedGoogle Scholar
• 17 Ozaki I, Tani E, Ikemoto H, Kitagawa H, Fujikawa H. Activation of stress-activated protein kinase/c-Jun NH2-terminal kinase and p38 kinase in calphostin C-induced apoptosis requires caspase-3 - like proteases but is dispensable for cell death.  J Biol Chem. 1999;  274 5310-7.
  CrossrefPubMedGoogle Scholar
• 18 Hansen M B, Nielsen S E, Berg K. Re-examination and further development of a precise and rapid dye method for measuring cell growth/cell kill.  J Immunol Methods. 1989;  119 203-10.
  CrossrefPubMedGoogle Scholar
• 19 Zhang Y G, Mattjus P, Schmid P C, Dong Z M, Zhong S P, Ma W Y. et al . Involvement of the acid sphingomyelinase pathway in UVA-induced apoptosis.  J Biol Chem. 2001;  276 11 775-82.
  PubMedGoogle Scholar
• 20 Boyce M, Degterev A, Yuan J. Caspases: an ancient cellular sword of Damocles.  Cell Death Differ. 2004;  11 29-37.
  CrossrefPubMedGoogle Scholar
• 21 Creagh E M, Martin S J. Caspases: cellular demolition experts.  Biochem Soc Trans. 2001;  29 696-701.
  CrossrefPubMedGoogle Scholar
• 22 Hussain S A, Ferry D R, EI Gazzaz G, Mirza D F, James N D, McMaster P. et al . Hepatocellular carcinoma.  Ann Oncol. 2001;  12 161-72.
  CrossrefPubMedGoogle Scholar
• 23 Suzuki T, Tsukamoto I. Arsenite induces apoptosis in hepatocytes through an enhancement of the activation of Jun N-terminal kinase and p38 mitogen-activated protein kinase caused by partial hepatectomy.  Toxicol Lett. 2006;  165 257-64.
  CrossrefPubMedGoogle Scholar
• 24 Corazza N, Jakob S, Schaer C, Frese S, Keogh A, Stroka D. et al . TRAIL receptor-mediated JNK activation and Bim phosphorylation critically regulate Fas-mediated liver damage and lethality.  J Clin Invest. 2006;  116 2493-9.
  PubMedGoogle Scholar
• 25 Bhakar A L, Howell J L, Christine E P, Salehi A H, Becker E BE, Said F. et al . Apoptosis induced by p75NTR overexpression requires Jun kinase-dependent phosphorylation of Bad.  J Neurosci. 2003;  23 11 373-81.
  PubMedGoogle Scholar
• 26 Kim B J, Ryu S W, Song B J. JNK- and p38 kinase-mediated phosphorylation of Bax leads to its activation and mitochondrial translocation and to apoptosis of human hepatoma HepG2 cells.  J Biol Chem. 2006;  281 21 256-65.
  PubMedGoogle Scholar
• 27 Yoo J, Ghiassi M, Jirmanova L, Balliet A G, Hoffman B, Fornace AJ J r. et al . Transforming growth factor-beta-induced apoptosis is mediated by Smad-dependent expression of GADD45b through p38 activation.  J Biol Chem. 2003;  278 43 001-7.
  PubMedGoogle Scholar
• 28 Young P R, McLaughlin M M, Kumar S, Kassis S, Doyle M L, McNulty D. et al . Pyridinyl imidazole inhibitors of p38 mitogen-activated protein kinase bind in the ATP site.  J Biol Chem. 1997;  272 12 116-21.
  PubMedGoogle Scholar
• 29 Huynh H, Nguyen T TT, Chow K HP, Tan P H, Soo K C, Tran E. Over-expression of the mitogen-activated protein kinase (MAPK) kinase (MEK)-MAPK in hepatocellular carcinoma: its role in tumor progression and apoptosis. BMC Gastroenterol 2003: 3.
  Google Scholar
• 30 Kar S, Wang M, Ham S W, Carr B I. H32, a non-quinone sulfone analog of vitamin K3, inhibits human hepatoma cell growth by inhibiting Cdc25 and activating ERK.  Cancer Biol Ther. 2006;  5 1340-7.
  PubMedGoogle Scholar

Prof. Dr. Zhengtao Wang

Key Laboratory of Standardization of Chinese Medicines of Education

Institute of Chinese Materia Medica

Shanghai University of Traditional Chinese Medicine

1200 Cai Lun Road

Shanghai 201203

People’s Republic of China

Phone: +86-21-5132-2507

Fax: +86-21-5132-2519

Email: wangzht@hotmail.com

Email: wangzht@shutcm.edu

• Supplementary Material