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Pretreatment Sensitizes Human Melanoma Cells to TRAIL/Apo2 Ligand-Induced Apoptosis1
Center for Drug Discovery and Development, Taussig Cancer Center, Cleveland Clinic Foundation, Cleveland, OH 44195
All human melanoma cell lines (assessed by annexin V and TUNEL
assays) were resistant to apoptosis induction by TRAIL/Apo2L protein.
TRAIL/Apo2L activated caspase-8 and caspase-3, but subsequent apoptotic
events such as poly(ADP-ribose) polymerase cleavage and DNA
fragmentation were not observed. To probe the molecular mechanisms of
cellular resistance to apoptosis, melanoma cell lines were analyzed for
expression of apoptosis regulators (apoptotic protease-associated
factor-1, FLIP, caspase-8, caspase-9, caspase-3, cellular inhibitor of
apoptosis, Bcl-2, or Bax); no correlation was observed. TRAIL/Apo2L was
induced in melanoma cell lines by IFN-
and had been correlated with
apoptosis induction. Because IFN- induced other gene products that
have been associated with apoptosis, it was postulated that one or more
IFN-stimulated genes might sensitize cells to TRAIL/Apo2L. Melanoma
cell lines were treated with IFN- for 16–24 h before treatment with
TRAIL/Apo2L. Regardless of their sensitivity to either cytokine alone,
>30% of cells underwent apoptosis in response to the combined
treatment. Induction of apoptosis by IFN- and TRAIL/Apo2L in
combination correlated with synergistic activation of caspase-9, a
decrease in mitochondrial potential, and cleavage of poly(ADP-ribose)
polymerase. Cleavage of X-linked inhibitor of apoptosis following
IFN- and TRAIL/Apo2L treatment was observed in sensitive WM9, A375,
or WM3211 cells but not in resistant WM35 or WM164 cells. Thus, in
vitro IFN- and TRAIL/Apo2L combination treatment had more potent
apoptotic and anti-growth effects when compared with either
cytokine alone in melanoma cells lines.
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