Axl and Vascular Endothelial Growth Factor Receptors Exhibit Variations in Membrane Localization and Heterogeneity Across Monolayer and Spheroid High-Grade Serous Ovarian Cancer Models
Axl and Vascular Endothelial Growth Factor Receptors Exhibit Variations in Membrane Localization and Heterogeneity Across Monolayer and Spheroid High-Grade Serous Ovarian Cancer Models
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DOI：
10.1089/genbio.2022.0034
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发表时间：
2023-02-01
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影响因子：
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通讯作者：
Imoukhuede, Princess I.
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文献类型：
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作者：
Fang, Yingye;Imoukhuede, Princess I.
血管内皮生长因子受体（VEGF）和Axl是在卵巢癌治疗中靶向的受体酪氨酸激酶（RTK）。二维单层培养和三维球状体是RTK靶向药物筛选的常见模型：单层简单且经济，而球状体包括几种遗传和组织学肿瘤特征。RTK膜定位决定了RTK信号传导和药物反应，然而，它在这些模型中没有表征。我们定量质膜RTK浓度，并显示差异RTK丰度和异质性在单层与球体。我们显示，OVCAR 8球状体中质膜上的VEGFR 1浓度比单层高10倍; OVCAR 8球状体比单层更不均匀，表现出低Axl（6200/细胞）和高Axl亚群（25，000/细胞）的双峰分布。此外，质膜Axl浓度在化学敏感性（OVCAR 3）和化学抗性（OVCAR 8）细胞之间相差100倍，并且在化学抗性细胞系之间相差10倍（OVCAR 5对OVCAR 8）。这些系统的发现可以指导卵巢癌模型的选择用于药物筛选。
Vascular endothelial growth factor receptors (VEGFRs) and Axl are receptor tyrosine kinases (RTK) that are targeted in ovarian cancer therapy. Two-dimensional monolayer culture and three-dimensional spheroids are common models for RTK-targeted drug screening: monolayers are simple and economical while spheroids include several genetic and histological tumor features. RTK membrane localization dictates RTK signaling and drug response, however, it is not characterized in these models. We quantify plasma membrane RTK concentrations and show differential RTK abundance and heterogeneity in monolayers versus spheroids. We show VEGFR1 concentrations on the plasma membrane to be 10 times higher in OVCAR8 spheroids than in monolayers; OVCAR8 spheroids are more heterogeneous than monolayers, exhibiting a bimodal distribution of a low-Axl (6200/cell) and a high-Axl subpopulation (25,000/cell). In addition, plasma membrane Axl concentrations differ by 100 times between chemosensitive (OVCAR3) and chemoresistant (OVCAR8) cells and by 10 times between chemoresistant cell lines (OVCAR5 vs. OVCAR8). These systematic findings can guide ovarian cancer model selection for drug screening.