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Generation of new hepatocyte-like in vitro models better resembling human lipid metabolism.
Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids ( IF 4.8 ) Pub Date : 2020-02-11 , DOI: 10.1016/j.bbalip.2020.158659
Camilla Pramfalk 1 , Tomas Jakobsson 2 , Cristy R C Verzijl 2 , Mirko E Minniti 2 , Clara Obensa 2 , Federico Ripamonti 2 , Maria Olin 2 , Matteo Pedrelli 2 , Mats Eriksson 3 , Paolo Parini 4
Affiliation  

In contrast to human hepatocytes in vivo, which solely express acyl-coenzyme A:cholesterol acyltransferase (ACAT) 2, both ACAT1 and ACAT2 (encoded by SOAT1 and SOAT2) are expressed in primary human hepatocytes and in human hepatoma cell lines. Here, we aimed to create hepatocyte-like cells expressing the ACAT2, but not the ACAT1, protein to generate a model that - at least in this regard - resembles the human condition in vivo and to assess the effects on lipid metabolism. Using the Clustered Regularly Interspaced Short Palindromic Repeats technology, we knocked out SOAT1 in HepG2 and Huh7.5 cells. The wild type and SOAT2-only-cells were cultured with fetal bovine or human serum and the effects on lipoprotein and lipid metabolism were studied. In SOAT2-only-HepG2 cells, increased levels of cholesterol, triglycerides, apolipoprotein B and lipoprotein(a) in the cell media were detected; this was likely dependent of the increased expression of key genes involved in lipid metabolism (e.g. MTP, APOB, HMGCR, LDLR, ACACA, and DGAT2). Opposite effects were observed in SOAT2-only-Huh7.5 cells. Our study shows that the expression of SOAT1 in hepatocyte-like cells contributes to the distorted phenotype observed in HepG2 and Huh7.5 cells. As not only parameters of lipoprotein and lipid metabolism but also some markers of differentiation/maturation increase in the SOAT2-only-HepG2 cells cultured with HS, this cellular model represent an improved model for studies of lipid metabolism.

中文翻译:

新型肝细胞样体外模型的产生更类似于人脂质代谢。

与体内仅表达酰基辅酶A:胆固醇酰基转移酶(ACAT)2的人肝细胞相比,ACAT1和ACAT2(由SOAT1和SOAT2编码)均在原代人肝细胞和人肝癌细胞系中表达。在这里,我们旨在创建表达ACAT2但不表达ACAT1蛋白质的肝细胞样细胞,以生成至少在这方面类似于人体状况的模型,并评估其对脂质代谢的影响。使用聚类的规则间隔的短回文重复序列技术,我们敲除了HepG2和Huh7.5细胞中的SOAT1。用胎牛或人血清培养野生型和仅SOAT2细胞,并研究其对脂蛋白和脂质代谢的影响。在仅SOAT2的HepG2细胞中,胆固醇,甘油三酸酯,检测到细胞培养基中的载脂蛋白B和脂蛋白(a);这可能取决于参与脂质代谢的关键基因(例如MTP,APOB,HMGCR,LDLR,ACACA和DGAT2)表达的增加。在仅SOAT2的Huh7.5细胞中观察到相反的作用。我们的研究表明,肝细胞样细胞中SOAT1的表达有助于在HepG2和Huh7.5细胞中观察到扭曲的表型。在用HS培养的SOAT2-only-HepG2细胞中,不仅脂蛋白和脂质代谢的参数增加,而且分化/成熟的一些标志物增加,这种细胞模型代表了脂质代谢研究的改进模型。在仅SOAT2的Huh7.5细胞中观察到相反的作用。我们的研究表明,肝细胞样细胞中SOAT1的表达有助于在HepG2和Huh7.5细胞中观察到扭曲的表型。在用HS培养的SOAT2-only-HepG2细胞中,不仅脂蛋白和脂质代谢的参数增加,而且分化/成熟的一些标志物增加,这种细胞模型代表了脂质代谢研究的改进模型。在仅SOAT2的Huh7.5细胞中观察到相反的作用。我们的研究表明,肝细胞样细胞中SOAT1的表达有助于在HepG2和Huh7.5细胞中观察到扭曲的表型。在用HS培养的SOAT2-only-HepG2细胞中,不仅脂蛋白和脂质代谢的参数增加,而且分化/成熟的一些标志物增加,这种细胞模型代表了脂质代谢研究的改进模型。
更新日期:2020-02-12
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