Synergistic Action of Genistein and Calcitriol in Immature Osteosarcoma MG-63 Cells by SGPL1 Up-Regulation.
Synergistic Action of Genistein and Calcitriol in Immature Osteosarcoma MG-63 Cells by SGPL1 Up-Regulation.
复制标题
SGPL1上调在未成熟骨肉瘤Mg-63细胞中染料木黄酮和钙三醇在未成熟骨肉瘤中的协同作用。
DOI：
10.1371/journal.pone.0169742
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Kraft K
中科院分区：
文献类型：
--
作者：
Engel N;Adamus A;Schauer N;Kühn J;Nebe B;Seitz G;Kraft K
植物雌激素，如染料木素（大豆中最重要的异黄酮），显示出浓度依赖性的抗雌激素或雌激素作用。高浓度金雀异黄素 (>10 μM) 也会促进体外骨癌细胞的增殖。另一方面，维生素 D 家族中最活跃的成分骨化三醇已被证明在体外和体内具有肿瘤保护作用。本研究的目的是检查金雀异黄素和骨化三醇在两种骨肉瘤细胞系 MG-63（早期成骨细胞）、Saos-2（成熟成骨细胞）和原代成骨细胞中的协同作用。因此，根据细胞周期时相改变、雌激素 (ER) 和维生素 D 受体 (VDR) 表达、活细胞代谢监测和代谢组学进行了初步筛选。暴露于 100 μM 金雀异黄素和 10 nM 骨化三醇的组合可将增殖细胞数量减少至对照水平，增加 ERß 和 VDR 表达，并减少细胞外酸化 (40%) 和呼吸活动 (70%)，主要是在 MG-63 细胞中。为了确定 MG-63 细胞系的潜在细胞机制，通过 GC/MS 技术进行代谢分析。联合治疗显着影响脂质和氨基酸，而能量代谢的代谢物没有改变。 log2比率的比较分析表明，联合治疗后，仅代谢物乙醇胺高度上调。结果是：1-磷酸鞘氨醇裂解酶 (SGPL1) 强烈过度表达 (350%)，它不可逆地降解 1-磷酸鞘氨醇 (S1P)，从而产生乙醇胺。 S1P 的产生和分泌与癌细胞迁移和侵袭能力的增强有关。从这些结果可以得出结论，高浓度金雀异黄素在未成熟骨肉瘤细胞中的肿瘤促进作用因骨化三醇的共同施用而降低，主要是通过S1P的分解而降低。应该在转移性异种移植小鼠模型中测试这种抗转移途径是否可以通过联合治疗来刺激。
Phytoestrogens such as genistein, the most prominent isoflavone from soy, show concentration-dependent anti-estrogenic or estrogenic effects. High genistein concentrations (>10 μM) also promote proliferation of bone cancer cells in vitro. On the other hand, the most active component of the vitamin D family, calcitriol, has been shown to be tumor protective in vitro and in vivo. The purpose of this study was to examine a putative synergism of genistein and calcitriol in two osteosarcoma cell lines MG-63 (early osteoblast), Saos-2 (mature osteoblast) and primary osteoblasts. Thus, an initial screening based on cell cycle phase alterations, estrogen (ER) and vitamin D receptor (VDR) expression, live cell metabolic monitoring, and metabolomics were performed. Exposure to the combination of 100 μM genistein and 10 nM calcitriol reduced the number of proliferative cells to control levels, increased ERß and VDR expression, and reduced extracellular acidification (40%) as well as respiratory activity (70%), primarily in MG-63 cells. In order to identify the underlying cellular mechanisms in the MG-63 cell line, metabolic profiling via GC/MS technology was conducted. Combined treatment significantly influenced lipids and amino acids preferably, whereas metabolites of the energy metabolism were not altered. The comparative analysis of the log2-ratios revealed that after combined treatment only the metabolite ethanolamine was highly up-regulated. This is the result: a strong overexpression (350%) of the enzyme sphingosine-1-phosphate lyase (SGPL1), which irreversibly degrades sphingosine-1-phosphate (S1P), thereby, generating ethanolamine. S1P production and secretion is associated with an increased capability of migration and invasion of cancer cells. From these results can be concluded that the tumor promoting effect of high concentrations of genistein in immature osteosarcoma cells is reduced by the co-administration of calcitriol, primarily by the breakdown of S1P. It should be tested whether this anti-metastatic pathway can be stimulated by combined treatment also in metastatic xenograft mice models.