Cinobufagin inhibits hepatocellular carcinoma EMT-like stemness via VEGF/VEGFR2 autocrine signaling.

IF 2.8 4区医学 Q3 ENDOCRINOLOGY & METABOLISM

Discover. Oncology Pub Date : 2025-05-26 DOI:10.1007/s12672-025-02707-9

Mingjun Ye, Shujun Chen, Chen Lu, Yangpei Wu, Jiaming Tian, Anqi Han, Jimin Zhu, Baikun Li, Qinglin Li

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Abstract

Purpose: This study aimed to explore the role of the VEGFa/VEGFR2 autocrine pathway in cinobufagin-induced inhibition of hepatocellular carcinoma metastasis.

Methods: A CCK-8 assay was performed to assess cell viability. Scratch healing, Transwell, and sphere formation assays were used to measure the effects of cinobufagin on cell migration, invasion, and tumor sphere formation. An immunofluorescence double staining method was used to detect the localization of VEGFa and VEGFR2. The effects of inhibiting the VEGFa/VEGFR2 autocrine pathway on Huh7 cell metastasis and the effects of the VEGFa/VEGFR2 autocrine pathway on the regulation of PI3K/AKT-dependent migration, invasion, and epithelial‒mesenchymal transition were examined through use of the VEGFR2 inhibitor apatinib and the PI3K inhibitor LY294002. The effects of cinobufagin on the VEGFa/VEGFR2 autocrine pathway and tumor metastasis were assessed in transplanted tumors.

Results: Cinobufagin inhibited Huh7 cell viability, migration, invasion, and tumor sphere formation in a dose-dependent manner. In addition, colocalization between VEGFa and VEGFR2 was detected in Huh7 cells. The results revealed that apatinib treatment significantly inhibited PI3K/AKT-dependent migration, invasion, and epithelial‒mesenchymal transition. The VEGFa/VEGFR2 autocrine pathway, the PI3K/AKT pathway, and epithelial-mesenchymal transition-associated protein markers were attenuated by cinobufagin in Huh7 cells. Additionally, cinobufagin attenuated the growth of hepatocellular carcinoma tumors in the Huh7 xenograft model and significantly downregulated the VEGFa/VEGFR2 autocrine pathway, the PI3K/AKT pathway, and the epithelial‒mesenchymal transition.

Conclusion: Cinobufagin may attenuate the PI3K/AKT-dependent metastatic potential of hepatocellular carcinoma by inhibiting the VEGFa/VEGFR2 autocrine pathway.

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Cinobufagin通过VEGF/VEGFR2自分泌信号抑制肝细胞癌emt样干细胞。

目的：本研究旨在探讨VEGFa/VEGFR2自分泌通路在朱诺费金诱导的肝细胞癌转移抑制中的作用。方法：采用CCK-8法测定细胞活力。使用划痕愈合、Transwell和球体形成试验来测量蟾毒球蛋白对细胞迁移、侵袭和肿瘤球体形成的影响。采用免疫荧光双染色法检测VEGFa和VEGFR2的定位。通过使用VEGFR2抑制剂阿帕替尼和PI3K抑制剂LY294002，研究了抑制VEGFa/VEGFR2自分泌途径对Huh7细胞转移的影响，以及VEGFa/VEGFR2自分泌途径对PI3K/ akt依赖性迁移、侵袭和上皮-间质转化的调节作用。评估cinobufagin对移植瘤中VEGFa/VEGFR2自分泌通路及肿瘤转移的影响。结果：Cinobufagin抑制Huh7细胞活力、迁移、侵袭和肿瘤球形成呈剂量依赖性。此外，在Huh7细胞中检测到VEGFa和VEGFR2的共定位。结果显示，阿帕替尼治疗显著抑制PI3K/ akt依赖性迁移、侵袭和上皮-间质转化。在Huh7细胞中，cinobufagin可减弱VEGFa/VEGFR2自分泌通路、PI3K/AKT通路和上皮-间质过渡相关蛋白标志物。此外，在Huh7异种移植模型中，cinobufagin可以抑制肝癌肿瘤的生长，并显著下调VEGFa/VEGFR2自分泌途径、PI3K/AKT途径和上皮-间质转化。结论：Cinobufagin可能通过抑制VEGFa/VEGFR2自分泌途径，减弱PI3K/ akt依赖性肝癌转移潜能。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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