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[Cancer Research 59, 4392-4399, September 1, 1999]
© 1999 American Association for Cancer Research
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[Cancer Research 59, 4392-4399, September 1, 1999]
© 1999 American Association for Cancer Research

Experimental Therapeutics

Hybrid Polar Histone Deacetylase Inhibitor Induces Apoptosis and CD95/CD95 Ligand Expression in Human Neuroblastoma1

Richard D. Glick2, Steven L. Swendeman2, Dennis C. Coffey, Richard A. Rifkind, Paul A. Marks, Victoria M. Richon and Michael P. La Quaglia3

Departments of Surgery [R. D. G., S. L. S., D. C. C., M. P. L.] and Cell Biology [R. A. R., P. A. M., V. M. R.], Sloan-Kettering Institute and Memorial Sloan-Kettering Cancer Center, New York, New York 10021

Inhibitors of histone deacetylase (HDAC) have been shown to have both apoptotic and differentiating effects on various tumor cells. M-carboxycinnamic acid bishydroxamide (CBHA) is a recently developed hybrid polar compound structurally related to hexamethylene bisacetamide. CBHA is a potent inhibitor of HDAC activity. CBHA induces cellular growth arrest and differentiation in model tumor systems. We undertook an investigation of the effects of CBHA on human neuroblastoma cell lines in vitro. When added to cultures of a panel of neuroblastoma cell lines, CBHA induced the accumulation of acetylated histones H3 and H4, consistent with the inhibition of HDAC. Concentrations of CBHA between 0.5 µM and 4 µM led to apoptosis in nine of nine neuroblastoma cell lines. Apoptosis was assessed by DNA fragmentation analysis and the appearance of a sub-G1 (<2N ploidy) population by flow cytometric analysis. The addition of a caspase inhibitor (benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone) completely abrogated CBHA-induced apoptosis in three of three cell lines. The addition of cycloheximide greatly reduced CBHA-induced apoptosis, suggesting that apoptotic induction was dependent on de novo protein synthesis. In addition, CBHA induced the expression of both CD95 (APO-1/Fas) and CD95 ligand within 12 h. The effect of CBHA on human neuroblastoma cells suggests that this agent and structurally related synthetic hybrid polar compounds have therapeutic potential for the treatment of this malignancy.




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