Min Junxia, Stegner Andrew L, Alexander Hannah, Alexander Stephen
Division of Biological Sciences, University of Missouri, Columbia, MO 65211-7400, USA.
Eukaryot Cell. 2004 Jun;3(3):795-805. doi: 10.1128/EC.3.3.795-805.2004.
The efficacy of the chemotherapy drug cisplatin is often limited due to resistance of the tumors to the drug, and increasing the potency of cisplatin without increasing its concentration could prove beneficial. A previously characterized Dictyostelium discoideum mutant with increased resistance to cisplatin was defective in the gene encoding sphingosine-1-phosphate (S-1-P) lyase, which catalyzes the breakdown of S-1-P, an important regulatory molecule in cell function and development and in the regulation of cell fate. We hypothesized that the increased resistance to cisplatin was due to an elevation of S-1-P and predicted that lowering levels of S-1-P should increase sensitivity to the drug. We generated three strains that stably overexpress different levels of the S-1-P lyase. The overexpressor strains have reduced growth rate and, confirming the hypothesis, showed an expression-dependent increase in sensitivity to cisplatin. Consistently, treating the cells with D-erythro-N,N,-dimethylsphingosine, a known inhibitor of sphingosine kinase, increased the sensitivity of mutant and parent cells to cisplatin, while addition of exogenous S-1-P or 8-Br-cyclic AMP made the cells more resistant to cisplatin. The increased sensitivity of the overexpressors to cisplatin was also observed with the cisplatin analog carboplatin. In contrast, the response to doxorubicin, 5-flurouracil, or etoposide was unaffected, indicating that the involvement of the sphingolipid metabolic pathway in modulating the response to cisplatin is not part of a global genotoxic stress response. The augmented sensitivity to cisplatin appears to be the result of an intracellular signaling function of S-1-P, because D. discoideum does not appear to have endothelial differentiation growth (EDG/S1P) receptors. Overall, the results show that modulation of the sphingolipid pathway at multiple points can result in increased sensitivity to cisplatin and has the potential for increasing the clinical usefulness of this important drug.
化疗药物顺铂的疗效常常因肿瘤对该药物产生耐药性而受到限制,在不增加顺铂浓度的情况下提高其效力可能会带来益处。先前鉴定出的一种对顺铂耐药性增强的盘基网柄菌突变体,其编码鞘氨醇-1-磷酸(S-1-P)裂解酶的基因存在缺陷,该酶催化S-1-P的分解,S-1-P是细胞功能、发育以及细胞命运调控中一种重要的调节分子。我们推测对顺铂耐药性增强是由于S-1-P水平升高所致,并预测降低S-1-P水平应会增加对该药物的敏感性。我们构建了三个稳定过表达不同水平S-1-P裂解酶的菌株。过表达菌株的生长速率降低,并且正如所推测的那样,显示出对顺铂的敏感性呈表达依赖性增加。同样,用已知的鞘氨醇激酶抑制剂D-赤藓醇-N,N-二甲基鞘氨醇处理细胞,可增加突变体细胞和亲本细胞对顺铂的敏感性,而添加外源性S-1-P或8-溴环磷酸腺苷则使细胞对顺铂更具耐药性。顺铂类似物卡铂也观察到过表达菌株对其敏感性增加。相比之下,对阿霉素、5-氟尿嘧啶或依托泊苷的反应未受影响,这表明鞘脂代谢途径参与调节对顺铂的反应并非整体遗传毒性应激反应的一部分。对顺铂敏感性增加似乎是S-1-P细胞内信号传导功能的结果,因为盘基网柄菌似乎没有内皮分化生长(EDG/S1P)受体。总体而言,结果表明在多个点调节鞘脂途径可导致对顺铂的敏感性增加,并有可能提高这种重要药物的临床效用。