Cancer and Translational Research Lab, Dr. D.Y. Patil Biotechnology & Bioinformatics Institute, Dr. D.Y. Patil Vidyapeeth, Pimpri, Pune, Maharashtra, India.
Bioinformatics Research Laboratory, Dr. D. Y. Patil Biotechnology and Bioinformatics Institute, Dr. D. Y. Patil Vidyapeeth, Pimpri, Pune, India.
Asian Pac J Cancer Prev. 2024 Feb 1;25(2):433-446. doi: 10.31557/APJCP.2024.25.2.433.
Cancer cells exhibit selective metabolic reprogramming to promote proliferation, invasiveness, and metastasis. Sphingolipids such as sphingosine and sphinganine have been reported to modulate cell death processes in cancer cells. However, the potential of extracellular sphinganine and its mimetic compounds as inducers of cancer cell death has not been thoroughly investigated.
We obtained extracellular conditioned medium from HCT-116 cells treated with the previously reported anticancer composition, goat urine DMSO fraction (GUDF). The extracellular metabolites were purified using a novel and in-house developed vertical tube gel electrophoresis (VTGE) technique and identified through LC-HRMS. Extracellular metabolites such as sphinganine, sphingosine, C16 sphinganine, and phytosphingosine were screened for their inhibitory role against intracellular kinases using molecular docking. Molecular dynamics (MD) simulations were performed to study the inhibitory potential of a novel designed modified mimetic sphinganine (MMS) (Pubchem CID: 162625115) upon c-Src kinase. Furthermore, inhibitory potential and ADME profile of MMS was compared with luteolin, a known c-Src kinase inhibitor.
Data showed accumulation of sphinganine and other sphingolipids such as C16 sphinganine, phytosphingosine, and ceramide (d18:1/14:0) in the extracellular compartment of GUDF-treated HCT-116 cells. Molecular docking projected c-Src kinase as an inhibitory target of sphinganine. MD simulations projected MMS with strong (-7.1 kcal/mol) and specific (MET341, ASP404) binding to the inhibitory pocket of c-Src kinase. The projected MMS showed comparable inhibitory role and acceptable ADME profile over known inhibitors.
In summary, our findings highlight the significance of extracellular sphinganine and other sphingolipids, including C16 sphinganine, phytosphingosine, and ceramide (d18:1/14:0), in the context of drug-induced cell death in HCT-116 cancer cells. Furthermore, we demonstrated the importance of extracellular sphinganine and its modified mimetic sphinganine (MMS) as a potential inhibitor of c-Src kinase. These findings suggest that MMS holds promise for future applications in targeted and combinatorial anticancer therapy.
癌细胞表现出选择性的代谢重编程，以促进增殖、侵袭和转移。已报道鞘脂如神经酰胺和神经酰胺具有调节癌细胞死亡过程的作用。然而，细胞外神经酰胺及其模拟化合物作为诱导癌细胞死亡的潜在物质尚未得到充分研究。
我们从用先前报道的抗癌组合物山羊尿 DMSO 馏分（GUDF）处理的 HCT-116 细胞中获得细胞外条件培养基。使用新型内部开发的垂直管凝胶电泳（VTGE）技术纯化细胞外代谢物，并通过 LC-HRMS 进行鉴定。筛选鞘氨醇、鞘氨醇、C16 神经酰胺和植物鞘氨醇等细胞外代谢物，通过分子对接研究其对细胞内激酶的抑制作用。进行分子动力学（MD）模拟研究新型设计的改良模拟神经酰胺（MMS）（Pubchem CID：162625115）对 c-Src 激酶的抑制潜力。此外，还比较了 MMS 的抑制潜力和 ADME 谱与已知的 c-Src 激酶抑制剂木犀草素。
数据显示，在 GUDF 处理的 HCT-116 细胞的细胞外隔室中积累了神经酰胺和其他鞘脂，如 C16 神经酰胺、植物鞘氨醇和神经酰胺（d18:1/14:0）。分子对接预测 c-Src 激酶是神经酰胺的抑制靶点。MD 模拟预测 MMS 与 c-Src 激酶的抑制口袋具有强烈的（-7.1 kcal/mol）和特异性（MET341、ASP404）结合。预测的 MMS 显示出与已知抑制剂相当的抑制作用和可接受的 ADME 谱。
总之，我们的研究结果强调了细胞外神经酰胺和其他鞘脂（包括 C16 神经酰胺、植物鞘氨醇和神经酰胺（d18:1/14:0））在 HCT-116 癌细胞药物诱导细胞死亡中的重要性。此外，我们还证明了细胞外神经酰胺及其修饰的模拟神经酰胺（MMS）作为 c-Src 激酶潜在抑制剂的重要性。这些发现表明，MMS 在靶向和组合抗癌治疗方面具有应用前景。
