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Abstract：
Background: This study aimed to examine the effect of 7-Ketositosterol (7-KSS), on sphingomyelin/ceramide metabolites and apoptosis in human breast MCF-7 and human liver HepG2 cancer cells. Methods: Anti-proliferative effects of 7-KSS treatment were assessed at different concentrations and periods. Cell viability was assessed through MTT analysis, whereas the levels of sphingosine-1-phosphate (S1P), sphingomyelins (SMs), and ceramides (CERs) were measured using LC-MS/MS. Phosphorylated 44/42 ERK1/2 and NF-κB p65 (Ser536) protein levels were measured by Western blot analysis and immunofluorescence staining. Apoptosis was evaluated by TUNEL staining and flow cytometric assessment of annexin-V and propidium iodide (PI) labeling. Results: Treatment with 7-KSS significantly decreased cell survival and S1P, p-44/42 ERK1/2, and p-NF-κB p65 protein levels in cancer cells compared to controls. A substantial rise was detected in intracellular amounts of C16-C24 CERs and apoptosis in cancer cells incubated with 7-KSS. Conclusions: 7-KSS stimulated ceramide accumulation and apoptosis while decreasing cell proliferation via downregulating S1P, p-44/42 ERK1/2, and p-NF-κB p65 protein levels.
摘要：
背景：本研究旨在研究7-酮甾醇（7-KSS）的作用，对人乳腺癌MCF-7和人肝癌HepG2细胞鞘磷脂/神经酰胺代谢产物和凋亡的影响。方法：评估7-KSS治疗在不同浓度和时期的抗增殖作用。通过MTT分析评估细胞活力，而鞘氨醇-1-磷酸（S1P）的水平，鞘磷脂（SM），使用LC-MS/MS测量神经酰胺（CER）。通过蛋白质印迹分析和免疫荧光染色测量磷酸化44/42ERK1/2和NF-κBp65（Ser536）蛋白水平。通过TUNEL染色和膜联蛋白V和碘化丙啶(PI)标记的流式细胞术评估来评估细胞凋亡。结果:7-KSS处理显著降低细胞存活率和S1P,与对照组相比，癌细胞中p-44/42ERK1/2和p-NF-κBp65蛋白水平。在与7-KSS孵育的癌细胞中，C16-C24CER的细胞内量和凋亡检测到了显着升高。结论：7-KSS通过下调S1P，刺激神经酰胺积累和凋亡，同时降低细胞增殖，p-44/42ERK1/2和p-NF-κBp65蛋白水平。
