Raj Ajay Kumar, Upadhyay Vidhi, Lokhande Kiran Bharat, Swamy K Venkateswara, Bhonde Ramesh Ramchandra, Sarode Sachin C, Sharma Nilesh Kumar
Cancer and Translational Research Lab, Dr. D. Y. Patil Biotechnology & Bioinformatics Institute, Dr. D. Y. Patil Vidyapeeth, Pimpri, Pune 411033, Maharashtra, India.
Bioinformatics Research Laboratory, Dr. D. Y. Patil Biotechnology and Bioinformatics Institute, Dr. D. Y. Patil Vidyapeeth, Pimpri, Pune 411033, Maharashtra, India.
Biomedicines. 2023 Mar 13;11(3):889. doi: 10.3390/biomedicines11030889.
The objective of this study was to explore the biological relevance of free fatty acids derived from cow urine DMSO fraction (CUDF) by employing in vitro and in silico approaches.
Metabolic heterogeneity at the intra- and intercellular levels contributes to the metabolic plasticity of cancer cells during drug-induced response. Free fatty acid (FFA) availability at intra- and intercellular levels is related to tumor heterogeneity at interpatient and xeno-heterogeneity levels.
We collected fresh urine from healthy cows and subjected it to fractionation in DMSO using drying, vortexing, and centrifugation. Finally, the sterile filtrate of cow urine DMSO fraction (CUDF) was evaluated for antiproliferative and proapoptotic effects in MCF-7 and ZR-75-1 breast cancer cells using routine cell-based assays. Intracellular metabolites were studied with the help of a novel in-house vertical tube gel electrophoresis (VTGE) method to reveal the nature of CUDF components in MCF-7 cells. Identified intracellular FFAs were studied for their molecular interactions with targeted receptor histone deacetylase (HDAC) using molecular docking and molecular dynamics (MD) simulations.
CUDF showed a significant reduction in cell viability and cell death in MCF-7 and ZR-75-1 breast cancer cells. Interestingly, FFAs tetracosanedioic acid, 13Z-docosenoic acid (erucic acid), nervonic acid, 3-hydroxy-tetradecanoic acid, and 3-hydroxcapric acid were found inside the treated MCF-7 cancer cells. These FFAs, including tetracosanedioic acid, indicated a specific affinity to HDAC at their inhibitory sites, similar to trichostatin A, a known inhibitor.
This study reports on FFAs derived from CUDF as potential antiproliferative and pro-cell death agents against breast cancer cells. MD simulations hinted at tetracosanedioic acid and other FFAs as inhibitors of HDAC that could explain the observed effects of FFAs in cancer cells.
本研究的目的是通过体外和计算机模拟方法探索源自牛尿二甲基亚砜馏分（CUDF）的游离脂肪酸的生物学相关性。
细胞内和细胞间水平的代谢异质性有助于癌细胞在药物诱导反应期间的代谢可塑性。细胞内和细胞间水平的游离脂肪酸（FFA）可用性与患者间肿瘤异质性和异种异质性水平相关。
我们收集了健康奶牛的新鲜尿液，并通过干燥、涡旋和离心在二甲基亚砜中进行分馏。最后，使用常规细胞检测方法评估牛尿二甲基亚砜馏分（CUDF）的无菌滤液对MCF-7和ZR-75-1乳腺癌细胞的抗增殖和促凋亡作用。借助一种新型的内部垂直管凝胶电泳（VTGE）方法研究细胞内代谢物，以揭示MCF-7细胞中CUDF成分的性质。使用分子对接和分子动力学（MD）模拟研究鉴定出的细胞内游离脂肪酸与靶向受体组蛋白脱乙酰酶（HDAC）的分子相互作用。
CUDF在MCF-7和ZR-75-1乳腺癌细胞中显示出细胞活力和细胞死亡的显著降低。有趣的是，在处理过的MCF-7癌细胞内发现了游离脂肪酸十四烷二酸、13Z-二十二碳烯酸（芥酸）、神经酸、3-羟基十四烷酸和3-羟基癸酸。这些游离脂肪酸，包括十四烷二酸，在其抑制位点对HDAC表现出特异性亲和力，类似于已知抑制剂曲古抑菌素A。
本研究报道了源自CUDF的游离脂肪酸作为针对乳腺癌细胞的潜在抗增殖和促细胞死亡剂。分子动力学模拟表明十四烷二酸和其他游离脂肪酸是HDAC的抑制剂，这可以解释游离脂肪酸在癌细胞中观察到的作用。
