Investigational New Drugs

, Volume 31, Issue 3, pp 525–534 | Cite as

Dual modulation of JNK and Akt signaling pathways by chaetoglobosin K in human lung carcinoma and ras-transformed epithelial cells

  • Amna Ali
  • Tatyana S. Sidorova
  • Diane F. Matesic


Chaetoglobosin K (ChK) is a natural product that inhibits anchorage-dependent and anchorage-independent growth of ras-transformed cells, prevents tumor-promoter disruption of cell-cell communication, and reduces Akt activation in tumorigenic cells. This study demonstrates how ChK modulates the JNK pathway in ras-transformed and human lung carcinoma cells and investigates regulatory mechanisms controlling ChK’s effect on the Akt and JNK signaling pathways. Human lung carcinoma and ras-transformed epithelial cell lines treated with ChK or vehicle for varying times were assayed for cell growth or extracted for total proteins for western blot analysis using phosphorylation site-specific antibodies to monitor changes in activation of JNK, Akt, and other signaling enzymes. Results show that ChK inhibited both Akt and JNK phosphorylation at key activation sites in ras-transformed cells as well as human lung carcinoma cells. Downstream effectors of both kinases were accordingly affected. Direct upstream kinases of JNK were not affected by ChK. Wortmannin and LY294002, two PI3 kinase inhibitors, inhibited Akt but not JNK phosphorylation in ras-transformed cells. This report establishes the dual inhibitory effect of ChK on both the Akt and JNK signaling pathways in ras-transformed epithelial and human carcinoma cells. The unique effect of ChK on these two key pathways involved in carcinogenesis earmarks ChK for further studies to determine its molecular target(s) and in vivo anti-tumor potential.


Chaetoglobosin K JNK Akt kinase Wortmannin LY294002 



This work was supported by the National Institute of Health, grant 1R15CA135415.

Conflict of interest

The authors declare that they have no conflict of interest.


  1. 1.
    Schechter AL, Stern DF, Vaidyanathan L, Decker SJ, Drebin JA, Greene MI et al (1984) The neu oncogene: an erb-B-related gene encoding a 185,000-Mr tumour antigen. Nature 312:513–516PubMedCrossRefGoogle Scholar
  2. 2.
    Jou YS, Layhe B, Matesic DF, Chang CC, de Feijter AW, Lockwood L et al (1995) Inhibition of gap junctional intercellular communication and malignant transformation of rat liver epithelial cells by neu oncogene. Carcinogenesis 16:311–317PubMedCrossRefGoogle Scholar
  3. 3.
    Adjei AA (2001) Blocking oncogenic Ras signaling for cancer therapy. J Natl Cancer Inst 93:1062–1074PubMedCrossRefGoogle Scholar
  4. 4.
    Jain M, Arvanitis C, Chu K, Dewey W, Leonhardt E, Trinh M et al (2002) Sustained loss of a neoplastic phenotype by brief inactivation of MYC. Science 297:102–104PubMedCrossRefGoogle Scholar
  5. 5.
    Druker BJ (2002) STI571 (Gleevec™) as a paradigm for cancer therapy. Trends Mol Med 8:S14–S18PubMedCrossRefGoogle Scholar
  6. 6.
    Bezjak A, Tu D, Seymour L, Clark G, Trajkovic A, Zukin M et al (2006) Symptom improvement in lung cancer patients treated with erlotinib: quality of life analysis of the National Cancer Institute of Canada Clinical Trials Group Study BR.21. J Clin Oncol 24:3831–3837PubMedCrossRefGoogle Scholar
  7. 7.
    Fukuoka M, Yano S, Giaccone G, Tamura T, Nakagawa K, Douillard JY et al (2003) Multi-institutional randomized phase II trial of gefitinib for previously treated patients with advanced non-small-cell lung cancer (The IDEAL 1 Trial) [corrected]. J Clin Oncol 21:2237–2246PubMedCrossRefGoogle Scholar
  8. 8.
    Kris MG, Natale RB, Herbst RS, Lynch TJ Jr, Prager D, Belani CP et al (2003) Efficacy of gefitinib, an inhibitor of the epidermal growth factor receptor tyrosine kinase, in symptomatic patients with non-small cell lung cancer: a randomized trial. JAMA 290:2149–2158PubMedCrossRefGoogle Scholar
  9. 9.
    Ross JS, Schenkein DP, Pietrusko R, Rolfe M, Linette GP, Stec J et al (2004) Targeted therapies for cancer 2004. Am J Clin Pathol 122:598–609PubMedCrossRefGoogle Scholar
  10. 10.
    Xiong HQ (2004) Molecular targeting therapy for pancreatic cancer. Cancer Chemother Pharmacol 54:S69–S77PubMedGoogle Scholar
  11. 11.
    Karapetis CS, Khambata-Ford S, Jonker DJ, O’Callaghan CJ, Tu D, Tebbutt NC et al (2008) K-ras mutations and benefit from cetuximab in advanced colorectal cancer. N Engl J Med 359:1757–1765PubMedCrossRefGoogle Scholar
  12. 12.
    Flaherty KT, Yasothan U, Kirkpatrick P (2011) Vemurafenib. Nat Rev Drug Discov 10:811–812PubMedCrossRefGoogle Scholar
  13. 13.
    Rodriguez-Viciana P, Warne PH, Khwaja A, Marte BM, Pappin D, Das P et al (1997) Role of phosphoinositide 3-OH kinase in cell transformation and control of the actin cytoskeleton by Ras. Cell 89:457–467PubMedCrossRefGoogle Scholar
  14. 14.
    Cantley LC (2002) The phosphoinositide 3-kinase pathway. Science 296:1655–1657PubMedCrossRefGoogle Scholar
  15. 15.
    Vivanco I, Sawyers CI (2002) The phosphatidylinositol 3-kinase-Akt pathway in human cancer. Nat Rev Cancer 2:489–501PubMedCrossRefGoogle Scholar
  16. 16.
    Stokoe D, Stephens LR, Copeland T, Gaffney PRJ, Reese CB, Painter GF et al (1997) Dual role of the phosphatidylinositol-3,4,5-trisphosphate in the activation of protein kinase B. Science 277:567–570PubMedCrossRefGoogle Scholar
  17. 17.
    Yang L, Dan HC, Sun M, Liu Q, Sun XM, Feldman RI et al (2004) Akt/protein kinase B signaling inhibitor-2, a selective small molecule inhibitor of Akt signaling with antitumor activity in cancer cells overexpressing Akt. Cancer Res 64:4394–4399PubMedCrossRefGoogle Scholar
  18. 18.
    Restuccia DF, Hemmings BA (2009) Blocking Akt-ivity. Science 325:1083–1084PubMedCrossRefGoogle Scholar
  19. 19.
    Sun M, Wang G, Paciga JE, Feldman RI, Yuan ZQ, Ma XL et al (2001) AKT1/PKBalpha kinase is frequently elevated in human cancers and its constitutive activation is required for oncogenic transformation in NIH3T3 cells. Am J Pathol 159:431–437PubMedCrossRefGoogle Scholar
  20. 20.
    Segrelles C, Ruiz S, Perez P, Murga C, Santos M, Budunova IV et al (2002) Functional roles of Akt signaling in mouse skin tumorigenesis. Oncogene 21:53–64PubMedCrossRefGoogle Scholar
  21. 21.
    Affara NI, Schanbacher BL, Mihm MJ, Cook AC, Pei P, Mallery SR et al (2004) Activated Akt-1 in specific cell populations during multi-stage skin carcinogenesis. Anticancer Res 24:2773–2782PubMedGoogle Scholar
  22. 22.
    Malik SN, Brattain M, Ghosh PM, Troyer DA, Prihoda T, Bedolla R et al (2002) Immunohistochemical demonstration of phospho-Akt in high Gleason grade prostate cancer. Clin Cancer Res 8:1168–1171PubMedGoogle Scholar
  23. 23.
    Kreisberg JI, Malik SN, Prihoda TJ, Bedolla RG, Troyer DA, Kreisberg S et al (2004) Phosphorylation of Akt (Ser473) is an excellent predictor of poor clinical outcome in prostate cancer. Cancer Res 64:5232–5236PubMedCrossRefGoogle Scholar
  24. 24.
    Antonyak MA, Kenyon LC, Godwin AK, James DC, Emlet DR, Okamoto I et al (2002) Elevated JNK activation contributes to the pathogenesis of human brain tumors. Oncogene 21:5038–5046PubMedCrossRefGoogle Scholar
  25. 25.
    Engelberg D (2004) Stress-activated protein kinases - tumor suppressors or tumor initiators? Semin Cancer Biol 14:271–282PubMedCrossRefGoogle Scholar
  26. 26.
    Khatlani TS, Wislez M, Sun M, Srinivas H, Iwanaga K, Ma L et al (2007) c-Jun N-terminal kinase is activated in non-small-cell lung cancer and promotes neoplastic transformation in human bronchial epithelial cells. Oncogene 26:2658–2666PubMedCrossRefGoogle Scholar
  27. 27.
    Cutler HG, Crumley F, Cox R (1980) Chaetoglobosin K: a new plant growth inhibitor and toxin from Diplodia macrospora. J Agric Food Chem 28:139–142PubMedCrossRefGoogle Scholar
  28. 28.
    Tikoo A, Cutler H, Lo SH, Chen LB, Maruta H (1999) Treatment of Ras-induced cancers by the F-actin cappers Tensin and Chaetoglobosin K, in combination with the Caspase-1 inhibitor N1445. Cancer J Sci Am 5:293–300PubMedGoogle Scholar
  29. 29.
    Matesic DF, Villio KN, Folse SL, Garcia EL, Cutler SJ, Cutler HG (2006) Inhibition of cytokinesis and Akt phosphorylation by chaetoglobosin K in ras-transformed epithelial cells. Cancer Chemother Pharmocol 57:741–754CrossRefGoogle Scholar
  30. 30.
    Matesic DF, Blommel ML, Sunman JA, Cutler SJ, Cutler HG (2001) Prevention of organochlorine-induced inhibition of gap junctional communication by chaetoglobosin K in astrocytes. Cell Biol Toxicol 17:395–408PubMedCrossRefGoogle Scholar
  31. 31.
    Sidorova TS, Matesic DF (2008) Protective effect of the natural product, Chaetoglobosin K, on lindane- and dieldrin-induced changes in astroglia: identification of activated signaling pathways. Pharm Res 25:1297–1308PubMedCrossRefGoogle Scholar
  32. 32.
    Engelman JA, Chen L, Tan X, Crosby K, Guimaraes AR, Upadhyay R et al (2008) Effective use of PI3K and MEK inhibitors to treat mutant Kras G12D and PIK3CA H1047R murine lung cancers. Nat Med 14:1351–1356PubMedCrossRefGoogle Scholar
  33. 33.
    Sarbassov DD, Guertin DA, Ali SM, Sabatini DM (2005) Phosphorylation and regulation of Akt/PKB by the rictor-mTOR complex. Science 307:1098–1101PubMedCrossRefGoogle Scholar
  34. 34.
    Kwon T, Kwon DY, Chun J, Kim JH, Kang SS (2000) Akt protein kinase inhibits Rac1-GTP binding through phosphorylation at serine 71 of Rac1. J Biol Chem 275:423–428PubMedCrossRefGoogle Scholar
  35. 35.
    Murga C, Zohar M, Teramoto H, Gutkind JS (2002) Rac1 and RhoG promote cell survival by the activation of PI3K and Akt, independently of their ability to stimulate JNK and NF-kappaB. Oncogene 21:207–216PubMedCrossRefGoogle Scholar
  36. 36.
    Zhang QG, Wang XT, Han D, Yin XH, Zhang GY, Xu TL (2006) Akt inhibits MLK3/JNK3 signaling by inactivating Rac1: a protective mechanism against ischemic brain injury. J Neurochem 98:1886–1898PubMedCrossRefGoogle Scholar
  37. 37.
    Logan MR, Mandato CA (2006) Regulation of the actin cytoskeleton by PIP2 in cytokinesis. Biol Cell 98:377–388PubMedCrossRefGoogle Scholar
  38. 38.
    Canman JC, Lewellyn L, Laband K, Smerdon SJ, Desai A, Bowerman B, Oegema K (2008) Inhibition of Rac by the GAP activity of centralspindlin is essential for cytokinesis. Science 322:1543–1546PubMedCrossRefGoogle Scholar
  39. 39.
    Lim CP, Cao X (1999) Serine phosphorylation and negative regulation of Stat3 by JNK. J Biol Chem 274:31055–31061PubMedCrossRefGoogle Scholar
  40. 40.
    Rawlings JS, Rosler KM, Harrison DA (2004) The JAK/STAT signaling pathway. J Cell Sci 117:1281–1283PubMedCrossRefGoogle Scholar
  41. 41.
    Wong MK, Lo AI, Lam B, Lam WK, Ip MS, Ho JC (2010) Erlotinib as salvage treatment after failure to first-line gefitinib in non-small cell lung cancer. Cancer Chemother Pharmacol 65:1023–1028PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2012

Authors and Affiliations

  • Amna Ali
    • 1
  • Tatyana S. Sidorova
    • 1
  • Diane F. Matesic
    • 1
  1. 1.Department of Pharmaceutical Sciences, College of Pharmacy and Health SciencesMercer UniversityAtlantaUSA

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