Fibrotic liver extracellular matrix induces cancerous phenotype in biomimetic micro-tissues of hepatocellular carcinoma model  

作　　者：Kosar Nouri Abbas Piryaei Homeyra Seydi Ibrahim Zarkesh Ibrahim Ghoytasi Bahare Shokouhian Mustapha Najimi Massoud Vosough 

机构地区：[1]Department of Developmental Biology,University of Science and Culture,ACECR 14155-4364 Tehran,Iran [2]Department of Regenerative Medicine,Cell Science Research Center,Royan Institute for Stem Cell Biology and Technology,ACECR 14155-4364 Tehran,Iran [3]Department of Stem Cells and Developmental Biology,Cell Science Research Center,Royan Institute for Stem Cell Biology and Technology,ACECR 14155-4364 Tehran,Iran [4]Department of Biology and Anatomical Sciences,School of Medicine,Shahid Beheshti University of Medical Sciences,Tehran,Iran [5]Department of Tissue Engineering and Applied Cell Sciences,School of Advanced Technologies in Medicine,Shahid Beheshti University of Medical Sciences,Tehran,Iran [6]Department of Cell Engineering,Cell Science Research Center,Royan Institute for Stem Cell Biology and Technology,ACECR 14155-4364 Tehran,Iran [7]Department of Medical Biotechnology,School of Advanced Technologies in Medicine,Tehran University of Medical Sciences,Tehran,Iran [8]Laboratory of Pediatric Hepatology and Cell Therapy,Institute of Experimental and Clinical Research(IREC),UCLouvain,Brussels,Belgium [9]Experimental Cancer Medicine,Institution for Laboratory Medicine,Karolinska Institute,Stockholm,Sweden

出　　处：《Hepatobiliary & Pancreatic Diseases International》2025年第1期92-103,共12页国际肝胆胰疾病杂志(英文版)

基　　金：financially supported by grants from Royan In-stitute(grant No.400000200);Bahar Tashkhis Teb Co.(BTT,9703,9809,and 9903)。

摘　　要：Background: Despite considerable advancements in identifying factors contributing to the development of hepatocellular carcinoma(HCC), the pathogenesis of HCC remains unclear. In many cases, HCC is a consequence of prolonged liver fibrosis, resulting in the formation of an intricate premalignant microenvironment. The accumulation of extracellular matrix(ECM) is a hallmark of premalignant microenvironment. Given the critical role of different matrix components in regulating cell phenotype and function, this study aimed to elucidate the interplay between the fibrotic matrix and malignant features in HCC. Methods: Liver tissues from both control(normal) and carbon tetrachloride(CCl_(4))-induced fibrotic rats were decellularized using sodium dodecyl sulfate(SDS) and Triton X-100. The resulting hydrogel from decellularized ECM was processed into micro-particles via the water-in-oil emulsion method. Microparticles were subsequently incorporated into three-dimensional liver biomimetic micro-tissues(MTs) comprising Huh-7 cells, human umbilical vein endothelial cells(HUVECs), and LX-2 cells. The MTs were evaluated using the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS) assay at day 11, immunofluorescence staining, immunoblotting, and spheroid migration assay at day 14 after co-culture. Results: Fibrotic matrix from CCl4-treated rat livers significantly enhanced the growth rate of the MTs and their expression of CCND1 as compared to the normal one. Fibrotic matrix, also induced the expression of epithelial-to-mesenchymal transition(EMT)-associated genes such as TWIST1, ACTA2, MMP9, CDH2, and VIMENTIN in the MTs as compared to the normal matrix. Conversely, the expression of CDH1 and hepatic maturation genes HNF4A, ALB, CYP3A4 was decreased in the MTs when the fibrotic matrix was used. Furthermore, the fibrotic matrix increased the migration of the MTs and their secretion of alpha-fetoprotein. Conclusions: Our findings suggest a regulatory role for the fibrotic matrix in promo

关 键 词：Liver fibrosis Decellularized extracellular matrix SPHEROID Epithelial-to-mesenchymal transition Cell differentiation Hepatocellular carcinoma 

分 类 号：R73[医药卫生—肿瘤]