Inhibition of phorbol ester-dependent differentiation of human promyelocytic leukemic (HL-60) cells by sphinganine and other long-chain bases.
Inhibition of phorbol ester-dependent differentiation of human promyelocytic leukemic (HL-60) cells by sphinganine and other long-chain bases.
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DOI：
10.1016/s0021-9258(18)67134-0
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发表时间：
1986-09
期刊：
影响因子：
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通讯作者：
A. Merrill;A. Sereni;V. Stevens;Y. Hannun;R. Bell;J. M. Kinkade
中科院分区：
文献类型：
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作者：
A. Merrill;A. Sereni;V. Stevens;Y. Hannun;R. Bell;J. M. Kinkade
研究了长链（类鞘氨醇）碱基对HL-60细胞佛波酯依赖性分化的影响，因为这些分子是蛋白激酶C的强效抑制剂（Hannun，Y.一、卢米斯角R.，梅里尔，A. H、小的，和贝尔，R. M.（1986）J.Biol.Chem.261，12604-12609）。24小时后，低浓度的鞘氨醇（1-5 μ M）阻断细胞粘附和响应佛波醇12-肉豆蔻酸酯13-乙酸酯的生长抑制，如通过细胞数量和酸性磷酸酶活性所测量的。鞘氨醇和鞘氨醇在1-3 μ M时可使粘附性降低50%;其他长链碱基在抑制蛋白激酶C的同时也有效。Sphinganine可降低HL-60细胞的佛波醇受体蛋白激酶C与[3 H]佛波醇二丁酸酯的结合，并抑制HL-60细胞对该酶的细胞渗透性激活剂二辛酰甘油的反应。用[3- 3 H]二氢鞘氨醇证明HL-60细胞的长链碱摄取，并且在1-3天内大部分已转化为神经酰胺。到第3天，大多数细胞已恢复粘附能力并表现出巨噬细胞特征，而悬浮液中的细胞不分化。HL-60细胞中游离鞘氨醇的水平测定为12.3 +/- 1.2 pmol/10（6）个细胞。这些结果表明，鞘氨醇碱抑制HL-60细胞中的蛋白激酶C，并可能作为这种酶的负效应生理功能。
The effects of long-chain (sphingoid) bases on the phorbol ester-dependent differentiation of HL-60 cells were investigated since these molecules are potent inhibitors of protein kinase C (Hannun, Y. A., Loomis, C. R., Merrill, A. H., Jr., and Bell, R. M. (1986) J. Biol. Chem. 261, 12604-12609). After 24 h, low concentrations of sphinganine (1-5 microM blocked both cell adherence and the inhibition of growth in response to phorbol 12-myristate 13-acetate, as measured by cell number and acid phosphatase activity. Sphinganine and sphingosine decreased adherence by 50% at 1-3 microM; other long-chain bases were effective in parallel to their inhibition of protein kinase C. Sphinganine decreased the binding of [3H]phorbol dibutyrate by the phorbol receptor of HL-60 cells, protein kinase C, and inhibited the response of HL-60 cells to dioctanoylglycerol, a cell permeable activator of this enzyme. Long-chain base uptake by HL-60 cells was demonstrated with [3-3H]sphinganine and within 1-3 days much had been converted to ceramides. By day 3, most of the cells had recovered the ability to adhere and exhibited macrophage characteristics, whereas cells in suspension did not differentiate. The level of free sphinganine in HL-60 cells was determined to be 12.3 +/- 1.2 pmol/10(6) cells. These results establish that sphingoid bases inhibit protein kinase C in HL-60 cells and may function physiologically as negative effectors of this enzyme.