The current study aims to evaluate the antiproliferative activity of B-norcholesteryl benzimidazole compounds in human ovarian cancer cells (SKOV3). Our experimental data indicates that the tested compounds can induce apoptosis in SKOV3 cells, block S-phase growth, and decrease mitochondrial membrane potential. Western blot results showed that B-norcholesteryl benzimidazole compounds (1 and 2) induced apoptosis in SKOV3 cells via activation of the mitochondrial signaling pathway. Following SKOV3 cells treatment with compounds 1 and 2, the cell metabolism was assessed using the UHPLC-QE-MS (Ultra High Performance Liquid Chromatography-Q Exactive Orbitrap- Mass Spectrometry) non-target metabolomics analysis method. The results showed 10 metabolic pathways that mediated the effects of compound 1, including arginine and proline metabolism; alanine, aspartate, and glutamate metabolism; histidine metabolism; D-glutamine and D-glutamine and D-glutamate metabolism; cysteine and methionine metabolism; aminoacyl-tRNA biosynthesis; purine metabolism; Glutathione metabolism; D-Arginine and D-ornithine metabolism; and Nitrogen metabolism. From the perspective of metabolomics, compound 1 inhibits intracellular metabolism, protein synthesis, and slows down energy metabolism in SKOV3 cells. These changes result in the inhibition of proliferation and signal transduction, abrogate invasive and metastatic properties, and induce apoptosis, thus, exerting anti-tumor effects. Application of compound 2 altered activation of metabolic pathways in SKOV3 cells. The main metabolic pathways involved were glycerophospholipid metabolism; arginine and proline metabolism; purine metabolism; glycine, serine, and threonine metabolism; and ether lipid metabolism. The metabolic pathway with the greatest impact and the deepest enrichment was the glycerophospholipid metabolism. In conclusion, compound 2 inhibits proliferation of SKOV3 cells by interfering with glycerate metabolism, which plays a major role in regulation of cell membrane structure and function. Additionally, compound 2 can inhibit the invasion and metastasis of SKOV3 cells and induce apoptosis via interfering with the metabolism of arginine and proline.

本研究旨在评估B-降胆固醇酯类苯并咪唑化合物在人卵巢癌细胞（SKOV3）中的抗增殖活性。我们的实验数据表明，所测试的化合物可以诱导SKOV3细胞凋亡，阻断S期生长并降低线粒体膜电位。Western印迹结果表明，B-降胆固醇基苯并咪唑化合物（1和2）通过激活线粒体信号传导途径诱导SKOV3细胞凋亡。用化合物1和2处理SKOV3细胞后，使用UHPLC-QE-MS（超高效液相色谱-Q Exactive Orbitrap-质谱）非目标代谢组学分析方法评估细胞代谢。结果表明，有10种代谢途径介导化合物1的作用，其中包括精氨酸和脯氨酸代谢。丙氨酸，天冬氨酸和谷氨酸代谢；组氨酸代谢 D-谷氨酰胺和D-谷氨酰胺和D-谷氨酸的代谢；半胱氨酸和蛋氨酸的代谢；氨酰基-tRNA的生物合成；嘌呤代谢；谷胱甘肽代谢；D-精氨酸和D-鸟氨酸代谢；和氮代谢。从代谢组学的角度看，化合物1抑制细胞内代谢，蛋白质合成，并减慢SKOV3细胞中的能量代谢。这些变化导致对增殖和信号转导的抑制，消除侵袭和转移特性，并诱导细胞凋亡，从而发挥抗肿瘤作用。化合物2的应用改变了SKOV3细胞中代谢途径的激活。涉及的主要代谢途径是甘油磷脂代谢。精氨酸和脯氨酸代谢；嘌呤代谢；甘氨酸，丝氨酸和苏氨酸代谢；和醚类脂代谢。影响最大，富集程度最大的代谢途径是甘油磷脂代谢。总之，化合物2通过干扰甘油酸酯代谢来抑制SKOV3细胞的增殖，甘油酸酯代谢在调节细胞膜结构和功能中起主要作用。另外，化合物2可通过干扰精氨酸和脯氨酸的代谢来抑制SKOV3细胞的侵袭和转移并诱导凋亡。