胸腺醌抑制血管生成能力并促进人乳腺癌干细胞的间质上皮转化。
Thymoquinone inhibited vasculogenic capacity and promoted mesenchymal-epithelial transition of human breast cancer stem cells.
机构信息
Student Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran.
Department of Biochemistry and Clinical Laboratories, Tabriz University of Medical Sciences, Tabriz, Iran.
出版信息
BMC Complement Med Ther. 2021 Mar 4;21(1):83. doi: 10.1186/s12906-021-03246-w.
BACKGROUND
Vasculogenic mimicry (VM) is characterized by the formation of tubular structure inside the tumor stroma. It has been shown that a small fraction of cancer cells, namely cancer stem cells (CSCs), could stimulate the development of vascular units in the tumor niche, leading to enhanced metastasis to the remote sites. This study aimed to study the inhibitory effect of phytocompound, Thymoquinone (TQ), on human breast MDA-MB-231 cell line via monitoring Wnt/PI3K signaling pathway.
METHODS
MDA-MB-231 CSCs were incubated with different concentrations of TQ for 48 h. The viability of CSCs was determined using the MTT assay. The combination of TQ and PI3K and Wnt3a inhibitors was examined in CSCs. By using the Matrigel assay, we measured the tubulogenesis capacity. The percent of CD24 CSCs and Rhodamine 123 efflux capacity was studied using flow cytometry analysis. Protein levels of Akt, p-Akt, Wnt3a, vascular endothelial-cadherin (VE-cadherin), and matrix metalloproteinases-2 and -9 (MMP-2 and -9) were detected by western blotting.
RESULTS
TQ decreased the viability of CSCs in a dose-dependent manner. The combination of TQ with PI3K and Wnt3a inhibitors reduced significantly the survival rate compared to the control group (p < 0.05). TQ could blunt the stimulatory effect of vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), fibroblast growth factor (FGF) on CSCs (p < 0.05). The vasculogenic capacity of CSCs was reduced after being-exposed to TQ (p < 0.05). Western blotting revealed the decrease of CSCs metastasis by suppressing MMP-2 and -9. The protein level of VE-cadherin was also diminished in TQ-treated CSCs as compared to the control cell (p < 0.05), indicating inhibition of mesenchymal-endothelial transition (MendT). TQ could suppress Wnt3a and PI3K, which coincided with the reduction of the p-Akt/Akt ratio. TQ had the potential to decrease the number of CD24 CSCs and Rhodamine 123 efflux capacity after 48 h.
CONCLUSION
TQ could alter the vasculogenic capacity and mesenchymal-epithelial transition of human breast CSCs in vitro. Thus TQ together with anti-angiogenic therapies may be a novel therapeutic agent in the suppression of VM in breast cancer.
背景
血管生成拟态（VM）的特征是肿瘤基质内管状结构的形成。已经表明，一小部分癌细胞，即癌症干细胞（CSC），可以刺激肿瘤龛位中血管单位的发育，导致远处转移的增强。本研究旨在通过监测 Wnt/PI3K 信号通路，研究植物化合物 Thymoquinone（TQ）对人乳腺癌 MDA-MB-231 细胞系的抑制作用。
方法
将 MDA-MB-231 CSC 与不同浓度的 TQ 孵育 48 小时。使用 MTT 测定法测定 CSC 的活力。在 CSC 中检查 TQ 与 PI3K 和 Wnt3a 抑制剂的组合。通过使用 Matrigel 测定法，我们测量了管形成能力。使用流式细胞术分析研究 CD24 CSC 的百分比和 Rhodamine 123 流出能力。通过 Western blot 检测 Akt、p-Akt、Wnt3a、血管内皮钙黏蛋白（VE-cadherin）和基质金属蛋白酶-2 和 -9（MMP-2 和 -9）的蛋白水平。
结果
TQ 以剂量依赖性方式降低 CSC 的活力。与对照组相比，TQ 与 PI3K 和 Wnt3a 抑制剂的组合显着降低了存活率（p<0.05）。TQ 可以削弱血管内皮生长因子（VEGF）、表皮生长因子（EGF）、成纤维细胞生长因子（FGF）对 CSC 的刺激作用（p<0.05）。CSC 的血管生成能力在暴露于 TQ 后降低（p<0.05）。Western blot 显示，通过抑制 MMP-2 和 -9，CSC 的转移减少。与对照细胞相比，TQ 处理的 CSC 中 VE-cadherin 的蛋白水平也降低（p<0.05），表明间充质-内皮转化（MendT）的抑制。TQ 可以抑制 Wnt3a 和 PI3K，这与 p-Akt/Akt 比值的降低一致。TQ 有可能在 48 小时后降低 CD24 CSC 的数量和 Rhodamine 123 流出能力。
结论
TQ 可以改变人乳腺癌 CSC 的血管生成能力和间充质上皮转化。因此，TQ 与抗血管生成治疗相结合可能是抑制乳腺癌中 VM 的新型治疗剂。
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