von Hippel-Lindau (VHL) disease, due to mutations of the tumor suppressorVHLgene, is a rare hereditary syndrome with a high risk of developing clear cell renal cell carcinoma (ccRCC). We asked whether the VHL-C162F mutation interferes with proliferation, migration, healing and forming colony ability by using wild-typeVHL(WT VHL) and VHL-C162F reconstituted cells. We then analyzed the in vitro impact of the sunitinib treatment on VHL-C162F cells. We showed that VHL-C162F mutations have no impact on cell morphology, colony formation and migration ability but confer a significant higher healing ability than in WT VHL cells. RNA sequencing analysis revealed that VHL-C162F mutation upregulates genes involved in hypoxia and epithelial mesenchymal transition (EMT) pathways by comparison with VHL WT cells. We next showed a decrease in healing ability in VHL-C162F cells depleting on ZHX2, an oncogenic driver of ccRCC, highlighting the potential involvement of ZHX2 in aggressiveness of the VHL-C162F cells. Moreover, we found that sunitinib treatment inhibits ZHX2 expression and induces a reduced proliferation correlating with downregulation of P-ERK. Sunitinib treatment also conferred a more mesenchymal profile to VHL-C162F cells with significant downregulation of E-cadherin and upregulation of N-cadherin, Slug and AXL. Sunitinib therapy may therefore promote disease progression in VHL-C162F patients.
von Hippel-Lindau (VHL) 病是一种罕见的遗传性综合征，由抑癌基因VHL突变引起，患者罹患透明细胞肾细胞癌 (ccRCC) 的风险极高。本研究通过使用野生型VHL (WT VHL) 和 VHL-C162F 重建细胞，探究 VHL-C162F 突变是否影响细胞的增殖、迁移、愈合及集落形成能力。随后，我们分析了舒尼替尼处理对 VHL-C162F 细胞的体外影响。结果显示，VHL-C162F 突变对细胞形态、集落形成和迁移能力无显著影响，但相较于 WT VHL 细胞，该突变赋予细胞显著更强的愈合能力。RNA 测序分析表明，与 VHL WT 细胞相比，VHL-C162F 突变上调了涉及缺氧和上皮间质转化 (EMT) 通路的相关基因。进一步研究发现，在敲低 ZHX2（一种 ccRCC 的致癌驱动因子）后，VHL-C162F 细胞的愈合能力下降，提示 ZHX2 可能参与该突变细胞的侵袭性表型。此外，舒尼替尼处理可抑制 ZHX2 表达，并通过下调 P-ERK 导致细胞增殖减少。同时，舒尼替尼处理使 VHL-C162F 细胞表现出更强的间质特征，具体表现为 E-钙黏蛋白显著下调，而 N-钙黏蛋白、Slug 和 AXL 显著上调。因此，舒尼替尼治疗可能促进 VHL-C162F 患者的疾病进展。
肿瘤(癌症)患者之家