Tumor Biology

, Volume 33, Issue 4, pp 1053–1058 | Cite as

RETRACTED ARTICLE: Calcium/calmodulin-dependent kinase inhibitor induces growth inhibition, cell cycle arrest, and apoptosis in human choriocarcinoma cells

  • Noriyuki Takai
  • Tami Ueda
  • Naoko Kira
  • Terukazu Ishii
  • Toshie Yoshida
  • Satoko Koga
  • Masakazu Nishida
  • Kaei Nasu
  • Hisashi Narahara
Research Article


KN-93, a membrane-permeant calcium/calmodulin-dependent kinase-selective inhibitor, induces apoptosis in some lines of human tumor cells. We investigated the effect of KN-93 in the choriocarcinoma cell line, BeWo. BeWo cells were treated with various concentrations of KN-93, and changes in cell growth, the cell cycle, apoptosis, and related parameters were examined. A WST-1 assay showed that BeWo cells were sensitive to the growth inhibitory effect of KN-93. Cell cycle analysis indicated that exposure to KN-93 decreased the proportion of cells in the S phase and increased the proportion in the G0/G1 phases of the cell cycle. Induction of apoptosis was confirmed by Annexin V staining of externalized phosphatidylserine, by the loss of mitochondrial transmembrane potential, and by antibodies directed against histones from fragmented DNA. This induction occurred in conjunction with the altered expression of genes related to cell growth, malignant phenotype, and apoptosis. These results suggest that KN-93 may serve as a therapeutic agent for the treatment of choriocarcinoma.


Calcium/calmodulin-dependent kinase KN-93 Cell cycle Apoptosis Choriocarcinoma 



This study was supported by a grant-in-aid (no. 21592139 to NT) for scientific research from the Ministry of Education, Culture, Sports, Science, and Technology, Japan, and a research fund at the discretion of the President, Oita University (HN).

Conflicts of interest



  1. 1.
    Jones WB, Cardinale C, Lewis Jr JL. Management of the high-risk gestational trophoblastic disease: the Memorial Hospital experience. Int J Gynecol Cancer. 1997;7:27–33.PubMedCrossRefGoogle Scholar
  2. 2.
    Berkowitz RS, Goldstein DP, Bernstein MR. Modified triple chemotherapy in the management of high-risk gestational trophoblastic tumors. Gynecol Oncol. 1984;19:173–81.PubMedCrossRefGoogle Scholar
  3. 3.
    Surwit EA, Hammond CB. Treatment of metastatic trophoblastic disease with poor prognosis. Obstet Gynecol. 1980;55:565–70.PubMedGoogle Scholar
  4. 4.
    Satsu H, Manabe M, Shimizu M. Activation of Ca2+/calmodulin-dependent protein kinase II is involved in hyperosmotic induction of the human taurine transporter. FEBS Lett. 2004;569:123–8.PubMedCrossRefGoogle Scholar
  5. 5.
    Rodriguez-Mora O, LaHair M, McCubrey J. FranklinR. Calcium/calmodulin-dependent kinase I and calcium/calmodulin-dependent kinase participate in the control of cell cycle progression in MCF-7 human breast cancer cells. Cancer Res. 2005;65:5408–16.PubMedCrossRefGoogle Scholar
  6. 6.
    Matsushita M, Nairn A. Characterization of the mechanism of regulation of Ca2+/calmodulin-dependent protein kinase I by calmodulin and by Ca2+/calmodulin-dependent protein kinase. J Biol Chem. 1998;273:21473–81.PubMedCrossRefGoogle Scholar
  7. 7.
    Tokumitsu H, Brickey D, Glod J, Hidaka H, Sikela J, Soderling T. Activation mechanisms for Ca2+/calmodulin-dependent protein kinase IV. Identification of a brain CaM-kinase IV kinase. J Biol Chem. 1994;269:28640–7.PubMedGoogle Scholar
  8. 8.
    Racioppi L, Means AR. Calcium/calmodulin-dependent kinase IV in immune and inflammatory responses: novel routes for an ancient traveler. Trends Immunol. 2008;29:600–7.PubMedCrossRefGoogle Scholar
  9. 9.
    Tombes RM, Grant S, Westin EH, Krystal G. G1 cell cycle arrest and apoptosis are induced in NIH 3T3 cells by KN-93, an inhibitor of CaMK-II (the multifunctional Ca2+/CaM kinase). Cell Growth Differ. 1995;6:1063–70.PubMedGoogle Scholar
  10. 10.
    Kawamura K, Grabowski D, Krivacic K, Hidaka H, Ganapathi R. Cellular events involved in the sensitization of etoposide-resistant cells by inhibitors of calcium–calmodulin-dependent processes. Role for effects on apoptosis, DNA cleavable complex, and phosphorylation. Biochem Pharmacol. 1996;52:1903–9.PubMedCrossRefGoogle Scholar
  11. 11.
    Sumi M, Kiuchi K, Ishikawa T, Ishii A, Hagiwara M, Nagatsu T, Hidaka H. The newly synthesized selective Ca2+/calmodulin dependent protein kinase II inhibitor KN-93 reduces dopamine contents in PC12h cells. Biochem Biophys Res Commun. 1991;181:968–75.PubMedCrossRefGoogle Scholar
  12. 12.
    Kahl C, Means A. Regulation of cyclin D1/Cdk4 complexes by calcium/calmodulin-dependent protein kinase I. J Biol Chem. 2004;279:15411–9.PubMedCrossRefGoogle Scholar
  13. 13.
    Shang S, Takai N, Nishida M, Miyazaki T, Nasu K, Miyakawa I. CaMKIV expression is associated with clinical stage and PCNA labeling index in endometrial carcinoma. Int J Mol Med. 2003;11:181–6.PubMedGoogle Scholar
  14. 14.
    Takai N, Ueda T, Nasu K, Yamashita S, Toyofuku M, Narahara H. Targeting calcium/calmodulin-dependence kinase I and II as a potential anti-proliferation remedy for endometrial carcinomas. Cancer Lett. 2009;277:235–43.PubMedCrossRefGoogle Scholar
  15. 15.
    Takai N, Desmond JC, Kumagai T, Gui D, Said JW, Whittaker S, et al. Histone deacetylase inhibitors have a profound anti-growth activity in endometrial cancer cells. Clin Cancer Res. 2004;10:1141–9.PubMedCrossRefGoogle Scholar
  16. 16.
    Takai N, Ueda T, Nishida M, Nasu K, Narahara H. Bufalin induces growth inhibition, cell cycle arrest and apoptosis in human endometrial and ovarian cancer cells. Int J Mol Med. 2008;21:637–43.PubMedGoogle Scholar
  17. 17.
    Rimon G, Bazenet CE, Philpott KL, Rubin LL. Increased surface phosphatidylserine is an early marker of neuronal apoptosis. J Neurosci Res. 1997;48:563–70.PubMedCrossRefGoogle Scholar
  18. 18.
    Chen Y, Kramer DL, Diegelman P, Vujcic S, Porter CW. Apoptotic signaling in polyamine analogue-treated SK-MEL-28 human melanoma cells. Cancer Res. 2001;61:6437–44.PubMedGoogle Scholar
  19. 19.
    Takai N, Ueda T, Nasu K, Narahara H. Erucylphosphocholine shows a strong anti-growth activity in human endometrial and ovarian cancer cells. Gynecol Oncol. 2008;111:336–43.PubMedCrossRefGoogle Scholar
  20. 20.
    Johnson D, Walker C. Cyclins and cell cycle checkpoints. Annu Rev Pharmacol Toxicol. 1999;39:295–312.PubMedCrossRefGoogle Scholar
  21. 21.
    Sherr C. G1 phase progression: cycling on cue. Cell. 1994;79:551–5.PubMedCrossRefGoogle Scholar
  22. 22.
    Kyushima N, Watanabe J, Hata H, Jobo T, Kameya T, Kuramoto H. Expression of cyclin A in endometrial adenocarcinoma and its correlation with proliferative activity and clinicopathological variables. J Cancer Res Clin Oncol. 2002;128:307–12.PubMedCrossRefGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2012

Authors and Affiliations

  • Noriyuki Takai
    • 1
  • Tami Ueda
    • 1
  • Naoko Kira
    • 1
  • Terukazu Ishii
    • 1
  • Toshie Yoshida
    • 1
  • Satoko Koga
    • 1
  • Masakazu Nishida
    • 1
  • Kaei Nasu
    • 1
  • Hisashi Narahara
    • 1
  1. 1.Department of Obstetrics and Gynecology, Faculty of MedicineOita UniversityOitaJapan

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