Skip to main content
Japanese Journal of Cancer Research : Gann logoLink to Japanese Journal of Cancer Research : Gann
. 1998 Sep;89(9):940–946. doi: 10.1111/j.1349-7006.1998.tb00652.x

Caspase‐3 Activation Is Not Responsible for Vinblastine‐induced Bcl‐2 Phosphorylation and G2/M Arrest in Human Small Cell Lung Carcinoma Ms‐1 Cells

Etsu Tashiro 1, Siro Simizu 1, Minoru Takada 2, Kazuo Umezawa 1, Masaya Imoto 1,
PMCID: PMC5921943  PMID: 9818030

Abstract

Vinblastine arrests cells in the G2/M phase of the cell cycle and subsequently induces cell death by apoptosis. We found that treatment of cells with vinblastine induced phosphorylation of Bcl‐2, resulting in the dissociation of Bcl‐2 and Bax. Moreover, vinblastine‐induced apoptosis was suppressed by an inhibitor of caspase‐3, Ac‐DEVD‐CHO; and a 17‐kDa active fragment of caspase‐3 was detected following vinblastine treatment, suggesting that caspase‐3 is involved in vinblastine‐induced apoptosis. However, Ac‐DEVD‐CHO affected neither vinblastine‐induced Bcl‐2 phosphorylation nor vinblastine‐induced G2/M arrest. Vinblastine caused G2/M arrest prior to apoptosis, whereas vinblastine‐induced apoptosis was not dependent on the duration of the G2/M phase. Thus, vinblastine‐induced apoptosis might be mediated by the phosphorylation of Bcl‐2, resulting in Bcl‐2 inactivation, and by subsequent activation of caspase‐3.

Keywords: key words, Vinblastine, Bcl‐2, Caspase‐3, G2/M arrest

Full Text

The Full Text of this article is available as a PDF (594.7 KB).

REFERENCES

  • 1. ) Jordan , M. A. , Wendell , K. , Gardiner , S. , Derry , W. B. , Copp , H. and Wilson , L.Mechanism of inhibition of cell proliferation by Vinca alkaloids . Cancer Res. , 51 , 2212 – 2222 ( 1991. ). [PubMed] [Google Scholar]
  • 2. ) Howard , S. M. H. , Theologides , A. and Sheppard , J. R.Comparative effects of vindesine, vinblastine, and vincristine on mitotic arrest and hormonal response of L1210 leukemia cells . Cancer Res. , 40 , 2695 – 2700 ( 1980. ). [PubMed] [Google Scholar]
  • 3. ) Wendell , K. L. , Wilson , L. and Jordan , M. A.Mitotic block in HeLa cells by vinblastine: ultrastructural changes in kinetochore‐microtubule attachment and in centrosomes . J. Cell Sci. , 104 , 261 – 272 ( 1993. ). [DOI] [PubMed] [Google Scholar]
  • 4. ) Donehower , L. A. and Bradley , A.The tumor suppressor p53 . Biochim. Biophys. Acta , 1155 , 181 – 205 ( 1993. ). [DOI] [PubMed] [Google Scholar]
  • 5. ) Zambetti , G. P. and Levine , A. J.A comparison of the biological activities of wild‐type and mutant p53 . FASEB J. , 7 , 855 – 865 ( 1993. ). [DOI] [PubMed] [Google Scholar]
  • 6. ) Yuan , J. , Shahan , S. , Lendoux , S. , Ellis , H. M. and Horvitz , H. R.The C. elegans cell death gene ced‐3 encodes a protein similar to mammalian interleukin‐1β‐converting enzyme . Cell , 75 , 641 – 652 ( 1993. ). [DOI] [PubMed] [Google Scholar]
  • 7. ) Lazebnik , Y. A. , Kaufmann , S. H. , Desnoyers , S. , Poirier , G. G. and Earnshaw , W. C.Cleavage of poly(ADP‐ribose) polymerase by a proteinase with properties like ICE . Nature , 371 , 346 – 347 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 8. ) Nicholson , D. W. , Ali , A. , Thornberry , N. A. , Vaillancourt , J. P. , Ding , C. K. , Gallant , M. , Gareau , Y. , Griffin , P. , Labelle , M. , Lazebnik , Y. A. , Munday , N. A. , Raju , S. M. , Smulson , M. E. , Yamin , T. T. , Yu , V. L. and Miller , D. K.Identification and inhibition of the ICE/CED‐3 protease necessary for mammalian apoptosis . Nature , 376 , 37 – 43 ( 1995. ). [DOI] [PubMed] [Google Scholar]
  • 9. ) Tewari , M. , Quan , L. T. , O'Rourke , K. , Desnoyers , S. , Zeng , Z. , Beidler , D. R. , Poirier , G. G. , Salvesen , G. S. and Dixit , V. M.Yama/CPP32β, a mammalian homolog of CED‐3, is a CrmA‐inhibitable protease that cleaves the death substrate poly(ADP‐ribose) polymerase . Cell , 81 , 801 – 809 ( 1995. ). [DOI] [PubMed] [Google Scholar]
  • 10. ) Hengartner , M. O. and Horvitz , H. R.Programmed cell death in Caenorhabditis elegans . Curr. Opin. Genet. Dev. , 4 , 581 – 586 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 11. ) Hengartner , M. O. and Horvitz , H. R.C. elegans cell‐survival gene Ced‐9 encodes a functional homolog of the mammalian protooncogene Bcl‐2 . Cell , 76 , 665 – 676 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 12. ) Chresta , C. M. , Masters , J. R. W. and Hickman , J. A.Hypersensitivity of human testicular tumors to etoposide‐induced apoptosis is associated with functional p53 and a high Bax:Bcl‐2 ratio . Cancer Res. , 56 , 1834 – 1841 ( 1996. ). [PubMed] [Google Scholar]
  • 13. ) Hockenbery , D. M. , Oltvai , Z. N. , Yin , X.‐M. , Milliman , C. L. and Korsmeyer , S. J.Bcl‐2 functions in an antioxidant pathway to prevent apoptosis . Cell , 75 , 241 – 251 ( 1993. ). [DOI] [PubMed] [Google Scholar]
  • 14. ) Kane , D. J. , Sarafian , T. A. , Anton , R. , Hahn , H. , Gralla , E. B. , Valentine , J. S. , Ord , T. and Bredesen , D. E.Bcl‐2 inhibition of neural death: decreased generation of reactive oxygen species . Science , 262 , 1274 – 1277 ( 1993. ). [DOI] [PubMed] [Google Scholar]
  • 15. ) Shimizu , S. , Eguchi , Y. , Kamiike , W. , Matsuda , H. and Tsujimoto , Y.Bcl‐2 expression prevents activation of the ICE protease cascade . Oncogene , 12 , 2251 – 2257 ( 1996. ). [PubMed] [Google Scholar]
  • 16. ) Ibrado , A. M. , Huang , Y. , Fang , G. and Bhalla , K.Bcl‐xL overexpression inhibits taxol‐induced Yama protease activity and apoptosis . Cell Growth Differ. , 7 , 1087 – 1094 ( 1996. ). [PubMed] [Google Scholar]
  • 17. ) Ibrado , A. M. , Huang , Y. , Fang , G. , Liu , L. and Bhalla , K.Overexpression of Bcl‐2 or Bcl‐xL inhibits Ara‐C‐induced CPP32/Yama protease activity and apoptosis of human acute myelogenous leukemia HL‐60 cells . Cancer Res. , 56 , 4743 – 4748 ( 1996. ). [PubMed] [Google Scholar]
  • 18. ) Guan , R. J. , Khatra , B. S. and Cohlberg , J. A.Phosphorylation of bovine neurofilament proteins by protein kinase FA (glycogen synthesis kinase 3) . J. Biol. Chem. , 266 , 8262 – 8267 ( 1991. ). [PubMed] [Google Scholar]
  • 19. ) Haldar , S. , Chintapalli , J. and Croce , C. M.Taxol induces bcl‐2 phosphorylation and death of prostate cancer cells . Cancer Res. , 56 , 1253 – 1255 ( 1996. ). [PubMed] [Google Scholar]
  • 20. ) Blagosklonny , M. , Schulte , T. , Nguyen , P. , Trepel , J. and Neckers , L. M.Taxol‐induced apoptosis and phosphorylation of Bcl‐2 protein involves c‐Raf‐1 and represents a novel c‐Raf‐1 signal transduction pathway . Cancer Res. , 56 , 1851 – 1854 ( 1996. ). [PubMed] [Google Scholar]
  • 21. ) Guan , R. J. , Moss , S. F. , Arber , N. , Krajewski , S. , Reed , J. C. and Holt , P. R.30 kDa phosphorylated form of Bcl‐2 protein in human colon . Oncogene , 12 , 2605 – 2609 ( 1996. ). [PubMed] [Google Scholar]
  • 22. ) Erhardt , P. and Cooper , G. M.Activation of the CPP32 apoptotic protease by distinct signaling pathways with differential sensitivity to Bcl‐xL . J. Biol. Chem. , 271 , 17601 – 17604 ( 1996. ). [DOI] [PubMed] [Google Scholar]
  • 23. ) Haldar , S. , Jena , N. and Croce , C. M.Inactivation of Bcl‐2 by phosphorylation . Proc. Natl. Acad. Sci. USA , 92 , 4507 – 4511 ( 1995. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 24. ) Haldar , S.Basu , A. and Croce , C. M.Bcl2 is the guardian of microtubule integrity . Cancer Res. , 57 , 229 – 233 ( 1997. ). [PubMed] [Google Scholar]
  • 25. ) Ibrado , A. M. , Liu , L. and Bhalla , K.Bcl‐XL overexpression inhibits progression of molecular events leading to paclitaxel‐induced apoptosis of human acute myeloid leukemia HL‐60 cells . Cancer Res. , 57 , 1109 – 1115 ( 1997. ). [PubMed] [Google Scholar]
  • 26. ) Blagosklonny , M. V. , Giannakakou , P. , El‐Deiry , W. S. , Kingston , D. G. I. , Higgs , P. I. , Neckers , L. and Fojo , T.Raf‐1/bcl‐2 phosphorylation: a step from microtubule damage to cell death . Cancer Res. , 57 , 130 – 135 ( 1997. ). [PubMed] [Google Scholar]
  • 27. ) Ito , T. , Deng , X. , Carr , B. and May , W. S.Bcl‐2 phosphorylation required for anti‐apoptosis function . J. Biol. Chem. , 272 , 11671 – 11673 ( 1997. ). [DOI] [PubMed] [Google Scholar]
  • 28. ) Xiang , J. , Chao , D. T. and Korsmeyer , S. J.Bax‐induced cell death may not require interleukin 1β‐converting enzyme‐like proteases . Proc. Natl. Acad. Sci. USA , 93 , 14559 – 14563 ( 1996. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 29. ) Simonian , P. L. , Grillot , D. A. M. , Andrews , D. W. , Leber , B. and Nunez , G.Bax homodimerization is not required for Bax to accelerate chemotherapy‐induced cell death . J. Biol. Chem. , 271 , 32073 – 32077 ( 1996. ). [DOI] [PubMed] [Google Scholar]

Articles from Japanese Journal of Cancer Research : Gann are provided here courtesy of Wiley

RESOURCES