N-acylated serinol is a novel ceramide mimic inducing apoptosis in neuroblastoma cells
N-acylated serinol is a novel ceramide mimic inducing apoptosis in neuroblastoma cells
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DOI：
10.1074/jbc.275.1.177
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发表时间：
2000-01-07
影响因子：
4.8
通讯作者：
Yu, RK
中科院分区：
文献类型：
--
作者：
Bieberich, E;Kawaguchi, T;Yu, RK
以丝氨醇（2-氨基-1，3-丙二醇）为原料，通过N-酰化反应合成了一种新的神经酰胺结构类似物，并研究了其对神经母细胞瘤细胞糖脂生物合成和细胞分化的影响。与N-棕榈酰化丝氨醇（C16-丝氨醇）孵育增加了内源性神经酰胺的浓度50-80%，并引起快速分裂的低密度细胞凋亡，但在汇合的细胞。与佛波酯同时孵育不能抑制细胞死亡，已知佛波酯通过激活蛋白激酶C（PKC）拮抗神经酰胺诱导的细胞凋亡。利用亲和层析法从大鼠脑中分离纯化了C16-丝氨醇的潜在靶蛋白。凝胶活性测定显示，从C16-serinol-Sepharose的酶包含三个丝氨酸/苏氨酸特异性蛋白激酶的分子量为50，70，和95 kDa。使用PKC ζ特异性抗体在Western印迹上对70-kDa蛋白进行免疫染色。在体外磷酸化试验中，纯化的PKC zeta可被C16-丝氨醇直接激活。Go 6983通过抑制PKC zeta抑制神经母细胞瘤细胞的凋亡诱导。我们的总体结果表明，C16-丝氨醇诱导的神经母细胞瘤细胞凋亡至少部分介导的PKC ζ的激活条件下进行细胞分裂。因此，N-酰化丝氨醇可用于诱导有丝分裂细胞中的凋亡，并且可具有治疗神经系统癌症的治疗潜力。
A novel structural analog of ceramide was synthesized by N-acylation of serinol (2-amino-1,3-propanediol) and studied for its effects on glycolipid biosynthesis and cell differentiation of neuroblastoma cells. Incubation with N-palmitoylated serinol (C16-serinol) increased the concentration of endogenous ceramide by 50-80% and caused apoptosis in rapidly dividing low density cells but not in confluent cells. Cell death was not suppressed by simultaneous incubation with phorbol ester, known to antagonize ceramide-induced apoptosis by activation of protein kinase C (PKC). Purification of potential target proteins of C16-serinol was achieved by affinity chromatography of a protein preparation from rat brain on immobilized C16-serinol. A gel activity assay revealed that the eluate from C16-serinol-Sepharose contained three serine/threonine-specific protein kinases with molecular masses of 50, 70, and 95 kDa. The 70-kDa protein was immunostained on a Western blot using a PKC zeta-specific antibody. The purified PKC zeta could be activated directly by C16-serinol in an in vitro phosphorylation assay. Induction of apoptosis in neuroblastoma cells was suppressed by inhibition of PKC zeta with Go 6983. Our overall results indicate that apoptosis in neuroblastoma cells induced by C16-serinol was at least partially mediated by activation of PKC zeta on condition of ongoing cell division. N-Acylated serinols may thus be useful for induction of apoptosis in mitotic cells and may be of therapeutic potential for treatment of cancer in the nervous system.