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Apoptosis

Clonal variability in CD95 expression is the major determinant in Fas-mediated, but not chemotherapy-mediated apoptosis in the RPMI 8226 multiple myeloma cell line

Leukemia volume 14, pages 830–840 (2000)Cite this article

Abstract

CD95 (Fas/APO-1) is a member of the TNFR superfamily that induces apoptosis following cross-linking with its cognate ligand, CD95L (FasL/APO-1L) or agonist antibody. The human myeloma cell line, RPMI 8226, has limited sensitivity to CD95-mediated apoptosis, with a maximum of 65% of the population responding. To determine the source of the limited sensitivity to CD95-mediated apoptosis, we isolated multiple clones from the RPMI-8226 cell line by limiting dilution. Analysis of these clones demonstrated that sensitivity to CD95-mediated cell death directly correlated with CD95 expression. Clones with high levels of CD95 expression had greater than 90% cell death, whereas cells with low levels of expression had less than 10% cell death. In contrast, no correlative differences were identified for other members of the DISC complex, or for members of the anti-apoptotic Bcl-2 family. We further examined the sensitivity of the 8226 clones to various cytotoxic agents. Although modest clonal variability was demonstrated in response to the chemotherapeutic drugs, doxorubicin, etoposide (VP-16), and vincristine, there was no correlation between CD95 function and sensitivity to chemotherapeutic drugs. These results indicate that in this cell line, receptor expression is rate limiting in CD95-mediated apoptosis, whereas CD95 expression was not a determinant in drug-induced programmed cell death.

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Acknowledgements

This study was supported in part by grants from the National Cancer Institute, CA77859 (WSD), core grant CA76292-01, and the flow cytometry and biostatistics core facilities at the H Lee Moffitt Center and Research Institute.

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Authors and Affiliations

  1. Clinical Investigations Program, H Lee Moffitt Cancer Center and Research Institute, University of South Florida, Tampa, FL, USA

    KH Shain, TH Landowski, I Buyuksal & WS Dalton

  2. Biostatistics Core, H Lee Moffitt Cancer Center and Research Institute, University of South Florida, Tampa, FL, USA

    AB Cantor

  3. Department of Biochemistry and Molecular Biology, University of South Florida, Tampa, FL, USA

    KH Shain & WS Dalton

  4. Department of Internal Medicine, University of South Florida, Tampa, FL, USA

    TH Landowski, I Buyuksal & WS Dalton

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Shain, K., Landowski, T., Buyuksal, I. et al. Clonal variability in CD95 expression is the major determinant in Fas-mediated, but not chemotherapy-mediated apoptosis in the RPMI 8226 multiple myeloma cell line. Leukemia 14, 830–840 (2000). https://doi.org/10.1038/sj.leu.2401776

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