A Novel Three-Dimensional Culture System: Decellularized-Tissue Obtained from Cirrhotic Livers Enhances an Epithelial-Mesenchymal Transition Phenotype in Hepatocellular Carcinoma Cells
A Novel Three-Dimensional Culture System: Decellularized-Tissue Obtained from Cirrhotic Livers Enhances an Epithelial-Mesenchymal Transition Phenotype in Hepatocellular Carcinoma Cells
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新型三维培养系统：从肝硬化肝脏获得的脱细胞组织增强肝细胞癌细胞的上皮-间质转化表型
DOI：
10.1016/j.jamcollsurg.2017.07.455
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发表时间：
2017
影响因子：
5.2
通讯作者：
Uemoto Shinji
中科院分区：
文献类型：
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作者：
Miyauchi Yuya;Yasuchika Kentaro;Fukumitsu Ken;Ishii Takamichi;Ogiso Satoshi;Kojima Hidenobu;Yamaoka Ryoya;Katayama Hokahiro;Kawai Takayuki;Uemoto Shinji
方法：雄性Lewis大鼠腹腔注射CCl4 8周，建立肝纤维化模型。采用脱细胞技术制备肝硬化肝支架。生成肝硬化肝脏支架后，将HCC细胞经胆管植入支架，培养7天。采用定量逆转录聚合酶链反应（qRT-PCR）分析HCC细胞的行为。结果：去细胞化的肝硬化肝脏胶原蛋白和硫酸氨基聚糖含量明显高于去细胞化的正常肝脏。去细胞化正常肝脏的HCC细胞进展为结节；去细胞化的肝硬化肝细胞癌以细胞外基质浸润的方式进展。qRT-PCR结果显示，去细胞化的肝硬化肝脏中，蜗牛、鼻涕虫、静脉蛋白和基质金属蛋白酶-9等间质标志物的表达明显高于去细胞化的正常肝脏。相反，与正常肝支架相比，肝硬化肝支架中上皮标志物E-cadherin的表达明显下调。结论：我们的新型三维培养系统保存了肝硬化肝脏的微环境。在去细胞化的肝硬化肝脏中培养的HCC细胞表现为上皮-间质转化表型。这种新的培养系统将有助于了解肝硬化的微环境和肝硬化的细胞生物学。
METHODS: Male Lewis rats were injected intraperitoneally with CCl4 for 8 weeks to establish the liver fibrosis model. Cirrhotic liver scaffolds were obtained using decellularized technique. After generating cirrhotic liver scaffolds, HCC cells were seeded into the scaffolds via the bile duct and cultured for 7 days. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was performed to analyze the behavior of HCC cells.RESULTS: The collagen and sulphated glycosaminoglycan content of the decellularized cirrhotic livers was significantly increased compared to that of decellularized normal livers. HCC cells in decellularized normal livers progressed to forming nodules; HCC cells in decellularized cirrhotic livers progressed with an infiltrating pattern between extracellular matrix. The qRT-PCR showed the expression of mesenchymal markers, such as snail, slug, vimentin, and matrix metalloproteinase-9, was significantly higher in decellularized cirrhotic livers than in decellularized normal livers. In contrast, expression of the epithelial marker, E-cadherin, in cirrhotic liver scaffolds was significantly downregulated compared to that in normal liver scaffolds.CONCLUSIONS: Our novel three-dimensional culture system was preserved the microenvironment of cirrhotic livers. HCC cells cultured in decellularized cirrhotic livers showed an epithelial-mesenchymal transition phenotype. This new culture system will be useful for understanding both microenvironment of cirrhotic livers and the cell biology in cirrhotic livers.