Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston, SC.
Laboratory of Biochemistry and Molecular Biology, San Paolo University Hospital, Medical School, University of Milan, Italy.
J Lipid Res. 2013 Mar;54(3):794-805. doi: 10.1194/jlr.M033985. Epub 2012 Nov 16.
Sphingomyelin synthase (SMS) produces sphingomyelin while consuming ceramide (a negative regulator of cell proliferation) and forming diacylglycerol (DAG) (a mitogenic factor). Therefore, enhanced SMS activity could favor cell proliferation. To examine if dysregulated SMS contributes to leukemogenesis, we measured SMS activity in several leukemic cell lines and found that it is highly elevated in K562 chronic myelogenous leukemia (CML) cells. The increased SMS in K562 cells was caused by the presence of Bcr-abl, a hallmark of CML; stable expression of Bcr-abl elevated SMS activity in HL-60 cells while inhibition of the tyrosine kinase activity of Bcr-abl with Imatinib mesylate decreased SMS activity in K562 cells. The increased SMS activity was the result of up-regulation of the Sms1 isoform. Inhibition of SMS activity with D609 (a pharmacological SMS inhibitor) or down-regulation of SMS1 expression by siRNA selectively inhibited the proliferation of Bcr-abl-positive cells. The inhibition was associated with an increased production of ceramide and a decreased production of DAG, conditions that antagonize cell proliferation. A similar change in lipid profile was also observed upon pharmacological inhibition of Bcr-abl (K526 cells) and siRNA-mediated down-regulation of BCR-ABL (HL-60/Bcr-abl cells). These findings indicate that Sms1 is a downstream target of Bcr-abl, involved in sustaining cell proliferation of Bcr-abl-positive cells.
鞘磷脂合酶 (SMS) 在消耗神经酰胺(细胞增殖的负调节剂)并形成二酰基甘油 (DAG)(有丝分裂原)的同时产生鞘磷脂。因此,增强 SMS 活性可能有利于细胞增殖。为了研究失调的 SMS 是否有助于白血病发生,我们测量了几种白血病细胞系中的 SMS 活性,发现 K562 慢性髓系白血病 (CML) 细胞中的 SMS 活性显著升高。K562 细胞中增加的 SMS 是由于存在 Bcr-abl,这是 CML 的标志;Bcr-abl 的稳定表达可提高 HL-60 细胞中的 SMS 活性,而使用伊马替尼抑制 Bcr-abl 的酪氨酸激酶活性则可降低 K562 细胞中的 SMS 活性。SMS 活性的增加是 Sms1 同工型上调的结果。使用 D609(一种药理学 SMS 抑制剂)抑制 SMS 活性或通过 siRNA 下调 SMS1 表达可选择性抑制 Bcr-abl 阳性细胞的增殖。这种抑制与神经酰胺产量增加和 DAG 产量减少有关,这两种情况都会拮抗细胞增殖。在药理学抑制 Bcr-abl(K526 细胞)和 siRNA 介导的 BCR-ABL 下调(HL-60/Bcr-abl 细胞)时,也观察到脂质谱发生类似变化。这些发现表明 Sms1 是 Bcr-abl 的下游靶标,参与维持 Bcr-abl 阳性细胞的增殖。