血管生成素（ANG）的分泌通过骨形态发生蛋白6（BMP6）/ID1信号通路，使内皮细胞在FIN56诱导的铁死亡性肝细胞癌中倾向于发生血管生成。

ANG secretion predisposes endothelial cells toward angiogenesis in FIN56-induced ferroptotic hepatocellular carcinoma via the BMP6/ID1 signaling pathway.

作者信息

Liu Xin, Long Gang, Wang Qingwen, Wang Jihong, Chen Xiaojing, Zhou Yuhang, You Kai, Li Han, Jiang Yi, Zhang Zhiyao, Du Xinrui, Han Yunwei

机构信息

Department of Oncology, The Affiliated Hospital, Southwest Medical University, Luzhou, 646000, China.

Department of General Surgery, The First Affiliated Hospital of Harbin Medical University, Harbin, 150007, China.

出版信息

Free Radic Biol Med. 2025 Jun 18;238:17-35. doi: 10.1016/j.freeradbiomed.2025.06.029.

DOI:10.1016/j.freeradbiomed.2025.06.029

PMID:40541804

Abstract

BACKGROUND

Hepatocellular carcinoma (HCC) poses significant challenges due to its high malignancy and limited treatment options. FIN56 has emerged as a potent inducer of ferroptosis, yet its precise mechanism of action in HCC remains elusive.

METHODS

Ferroptosis induction by FIN56 in HCC cells was assessed by quantifying malondialdehyde (MDA), 4-hydroxynonenal (4-HNE), reactive oxygen species (ROS), glutathione (GSH) and Liperfluo levels, as well as evaluating changes in mitochondrial morphology. Additionally, cell proliferation and migration assays were performed to assess the functional impact of FIN56 on HCC cells. Tube formation and sprouting assays were performed using human umbilical vein endothelial cells (HUVECs). Proteomics analysis and immunosorbent assay (ELISA) identified angiogenin (ANG) as a secreted protein in the supernatant of HCC cells. Furthermore, both xenograft and syngeneic tumor models were established to investigate the potential impact of FIN56-induced ferroptosis on the tumor microenvironment.

RESULTS

RNA sequencing analysis of FIN56-treated HCC cells identified differentially expressed genes mainly associated with ferroptosis, cell proliferation, and migration. FIN56 effectively induced ferroptosis in HCC cells, while simultaneously inhibiting their proliferation and migration. RNA sequencing of HUVECs exposed to conditioned medium (CM) from FIN56-treated HCC cells (FIN56-CM) showed significant alterations in endothelial cell growth and gene expression. Further experiments revealed that FIN56-treated HCC cells secreted substantial levels of ANG after 12-18h, which activated the BMP6/ID1 signaling axis in endothelial cells, thereby enhancing their tube formation. Notably, the BMP signaling inhibitor LDN214117 partially suppressed ANG-mediated effects on endothelial cells. Finally, both ferroptosis and pro-angiogenic effects of FIN56 were confirmed in xenograft tumor model.

CONCLUSION

FIN56 induces ferroptosis in HCC cells, leading to the secretion of ANG. The secreted ANG acts as a critical signaling molecule, activating the BMP6/ID1 pathway in HUVECs and contributing to tumor microenvironment regulation.

摘要

背景

肝细胞癌（HCC）因其高恶性和有限的治疗选择而带来重大挑战。FIN56已成为铁死亡的有效诱导剂，但其在HCC中的精确作用机制仍不清楚。

方法

通过定量丙二醛（MDA）、4-羟基壬烯醛（4-HNE）、活性氧（ROS）、谷胱甘肽（GSH）和Liperfluo水平，以及评估线粒体形态的变化，来评估FIN56在HCC细胞中诱导铁死亡的情况。此外，进行细胞增殖和迁移试验以评估FIN56对HCC细胞的功能影响。使用人脐静脉内皮细胞（HUVECs）进行管形成和发芽试验。蛋白质组学分析和免疫吸附测定（ELISA）确定血管生成素（ANG）为HCC细胞上清液中的一种分泌蛋白。此外，还建立了异种移植和同基因肿瘤模型，以研究FIN56诱导的铁死亡对肿瘤微环境的潜在影响。

结果

对经FIN56处理的HCC细胞进行RNA测序分析，确定了主要与铁死亡、细胞增殖和迁移相关的差异表达基因。FIN56有效地诱导了HCC细胞中的铁死亡，同时抑制了它们的增殖和迁移。对暴露于经FIN56处理的HCC细胞（FIN56-CM）的条件培养基（CM）中的HUVECs进行RNA测序，显示内皮细胞生长和基因表达有显著改变。进一步的实验表明，经FIN56处理的HCC细胞在12-18小时后分泌大量的ANG，其激活了内皮细胞中的BMP6/ID1信号轴，从而增强其管形成。值得注意的是，BMP信号抑制剂LDN214117部分抑制了ANG对内皮细胞的作用。最后在异种移植肿瘤模型中证实了FIN56的铁死亡和促血管生成作用。