Activation of caspase-3 in renal cell carcinoma cells by anthracyclines or 5-fluorouracil

  • Authors:
    • Xiu-Xian Wu
    • Y. Kakehi
    • Y. Mizutani
    • J. Lu
    • T. Terachi
    • O. Ogawa
  • View Affiliations

  • Published online on: July 1, 2001     https://doi.org/10.3892/ijo.19.1.19
  • Pages: 19-24
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Abstract

The caspase family of proteases is speculated to have a crucial role in apoptosis. The effect of treatment with adriamycin (ADR), cisplatin (CDDP), 5-fluorouracil (5-FU), vinblastine (VLB), IFN-α, or IFN-γ on the activation of caspase-3, -6, -8, and -9 in renal cell carcinoma (RCC) cells was investigated, to clarify the mechanisms of chemo- and immunotherapeutic agent-mediated apoptosis. Caspase activity was determined by a quantitative colorimetric assay. Apoptosis was monitored by acridine-orange staining assay. Treatment of ACHN cells with CDDP, VLB, IFN-α, or IFN-γ did not activate caspase-3, but its activity was increased 7.2-fold (p=0.0001) with ADR and 2.8-fold (p=0.0385) with 5-FU in comparison with control. Furthermore, when the ADR treatment time was shortened from 24 to 8 or 2 h, the same caspase-3 activation occurred. Activation of caspase-3 was also observed in six freshly isolated human RCC cells after the treatment with ADR. Of the six freshly derived RCC cells treated with 5-FU, caspase-3 activity was increased 3.1-fold (p=0.0051) and 2.4-fold (p=0.0346) in two of them, respectively. Epirubicin and pirarubicin, compounds closely related to ADR, also respectively enhanced 4.2-fold (p=0.0052) and 2.8-fold (p=0.0147) caspase-3 activity in ACHN cells. The activation of caspase-3 observed with a colorimetric assay was confirmed with immunocytochemical analysis using the anti-active caspase-3 mAb, which specifically recognizes the active form of caspase-3. Furthermore, both active caspase-3 and apoptosis triggered by either ADR or 5-FU were inhibited significantly by the general caspase inhibitor Z-VAD-FMK, or a specific caspase-3 inhibitor DMQD-CHO. These findings provide a mechanistic explanation for anthracyclines and 5-FU induced-apoptosis.

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July 2001
Volume 19 Issue 1

Print ISSN: 1019-6439
Online ISSN:1791-2423

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Spandidos Publications style
Wu X, Kakehi Y, Mizutani Y, Lu J, Terachi T and Ogawa O: Activation of caspase-3 in renal cell carcinoma cells by anthracyclines or 5-fluorouracil. Int J Oncol 19: 19-24, 2001
APA
Wu, X., Kakehi, Y., Mizutani, Y., Lu, J., Terachi, T., & Ogawa, O. (2001). Activation of caspase-3 in renal cell carcinoma cells by anthracyclines or 5-fluorouracil. International Journal of Oncology, 19, 19-24. https://doi.org/10.3892/ijo.19.1.19
MLA
Wu, X., Kakehi, Y., Mizutani, Y., Lu, J., Terachi, T., Ogawa, O."Activation of caspase-3 in renal cell carcinoma cells by anthracyclines or 5-fluorouracil". International Journal of Oncology 19.1 (2001): 19-24.
Chicago
Wu, X., Kakehi, Y., Mizutani, Y., Lu, J., Terachi, T., Ogawa, O."Activation of caspase-3 in renal cell carcinoma cells by anthracyclines or 5-fluorouracil". International Journal of Oncology 19, no. 1 (2001): 19-24. https://doi.org/10.3892/ijo.19.1.19