IGFBP-7 transforms hepatic stellate cells into an HCC-promoting phenotype in MASLD.

Hepatic fibrosis is the strongest contributor to hepatocarcinogenesis in metabolic dysfunction-associated steatotic liver disease (MASLD); however, the underlying mechanisms have yet to be fully elucidated. In 94 human MASLD biopsy samples, artificial intelligence-based morphological phenotyping of hepatic fiber and multi-omics analyses revealed that insulin growth factor-binding protein 7 (IGFBP-7) secreted from senescent periportal endothelial cells might transform stellate cells into a hepatocarcinogenesis-promoting phenotype. To test the effect of IGFBP-7 on HSC, a hepatic stellate cell line, LX-2, was cultured with recombinant IGFBP-7 (100ng/mL), resulting in their transformation to a more activated form than the control. Overall design: Hepatic stellate cell (HSC) line LX-2 (Merck Sigma-Aldrich, #SCC064) was cultured with DMEM and 2% fetal bovine serum (FBS). After incubation without FBS for 18 hours, cells were treated with IGFBP-7 (100ng/mL, R&D, #1334-B7) or phosphate-buffered saline and were incubated for an additional 24 hours. The total RNA of the treated cells was extracted using TRI Reagent (Sigma-Aldrich) and subjected to RNA sequencing. Library preparation was performed using NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (New England Biolabs) and sequencing was performed with NovaSeq 6000, according to the manufacturer's instructions.

在代谢功能障碍相关性脂肪性肝病（metabolic dysfunction-associated steatotic liver disease, MASLD）患者中，肝纤维化是肝细胞癌变发生的最主要驱动因素，但其潜在分子机制尚未完全阐明。研究人员对94例人类MASLD活检样本开展了肝纤维的人工智能辅助形态表型分析与多组学分析，结果发现：衰老门管区内皮细胞分泌的胰岛素样生长因子结合蛋白7（insulin growth factor-binding protein 7, IGFBP-7）可将肝星状细胞转化为促肝细胞癌变表型。为验证IGFBP-7对肝星状细胞（hepatic stellate cell, HSC）的作用，研究人员采用重组IGFBP-7（100ng/mL）处理肝星状细胞系LX-2，结果显示其较对照组细胞转化为更活化的表型。实验整体设计：肝星状细胞系LX-2（默克Sigma-Aldrich，货号#SCC064）采用DMEM培养基与2%胎牛血清（fetal bovine serum, FBS）进行培养。经无FBS孵育18小时后，分别用IGFBP-7（100ng/mL，R&D，货号#1334-B7）或磷酸盐缓冲液处理细胞，继续孵育24小时。使用TRI Reagent（Sigma-Aldrich）提取处理后细胞的总RNA，随后进行RNA测序。采用NEBNext Ultra II Directional RNA Library Prep Kit for Illumina（纽英伦生物技术，New England Biolabs）完成文库构建，并按照试剂盒说明书使用NovaSeq 6000平台完成测序。