© 1997 by Oxford University Press
Journal Of The National Cancer Institute, Vol 89, 783-789, Copyright © 1997 by Oxford University Press
O Micheau, E Solary, A Hammann, F Martin and MT Dimanche-Boitrel
BACKGROUND: The transmembrane receptor Fas, together with its protein-
binding partner (Fas ligand), is a key regulator of programmed cell death
(i.e., apoptosis). Fas and Fas ligand also influence the ability of
cytotoxic T lymphocytes and natural killer cells to eliminate tumor cells.
However, by inducing apoptosis in activated T cells, the Fas/Fas ligand
system may protect some tumor cells from clearance by the immune system.
Anticancer drugs enhance Fas ligand expression on the surface of Fas
receptor-expressing leukemia cells, thus suggesting that apoptosis caused
by these drugs may be mediated via the Fas/Fas ligand system. PURPOSE: This
study was conducted to further investigate the relationship between the
modulation of Fas receptor gene and protein expression by treatment of
cells with cytotoxic drugs and the immune clearance of tumor cells.
METHODS: Fas expression on human HT29 colon carcinoma cells treated with a
variety of anticancer drugs (cisplatin, doxorubicin, mitomycin C,
fluorouracil, and camptothecin) was analyzed by use of quantitative flow
cytometry. Human HCT8R and HCT116 colon carcinoma cells and human U937
leukemia cells were treated with cisplatin only and analyzed in the same
way. Fas ligand messenger RNA and protein levels were studied by use of a
reverse transcription- polymerase chain reaction assay and by flow
cytometry. Fas gene expression and messenger RNA levels in
cisplatin-treated HT29 cells were characterized by use of in vitro nuclear
run-on and northern blot hybridization assays. The cytotoxic activities of
agonistic anti-Fas antibodies, Fas ligand, and allogeneic peripheral blood
leukocytes, in the absence or presence of Fas-blocking monoclonal
antibodies, against tumor cells were assessed by methylene blue staining
and chromium-51 release assays. RESULTS: Clinically relevant concentrations
of cisplatin, doxorubicin, mitomycin C, fluorouracil, or camptothecin
enhanced Fas receptor expression on the plasma membrane of HT29 cells.
Cisplatin-mediated increases in Fas expression were confirmed in HCT8R,
HCT116, and U937 cells. The enhancement of Fas protein expression was
associated with an increased sensitivity of cisplatin-treated tumor cells
to agonistic anti-Fas antibodies, to soluble Fas ligand, and to allogeneic
peripheral blood leukocyte-mediated cytotoxicity. Each of these effects was
blocked by co-treatment of the cells with antagonistic anti-Fas antibody.
CONCLUSION AND IMPLICATIONS: In addition to their direct cytotoxic effects,
chemotherapeutic drugs sensitize tumor cells to Fas-mediated cytotoxicity
and Fas-dependent immune clearance. On the basis of these findings, new
strategies might be developed to improve the efficacy of these drugs.
ARTICLES
Sensitization of cancer cells treated with cytotoxic drugs to fas- mediated cytotoxicity
Contrat Jeune Formation de l'Institut National de la Sante et de la Recherche Medicale (INSERM) 94-08, Unite de Formation et de Recherchede Medecine, Dijon, France.
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