Reversible conversion of epithelial and mesenchymal phenotypes in SV40 large T antigen-immortalized rat liver cell lines

Takato Takenouchi, Miyako Yoshioka, Noriko Yamanaka, Hiroshi Kitani
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引用次数: 11

Abstract

EMT (epithelial—mesenchymal transition) is a key process in the development of liver fibrosis. This process is also essential for liver morphogenesis in embryonic development. To study the cellular and molecular basis of EMT, we established two phenotypically different SV40 large T antigen-immortalized cell lines from rat hepatocytes. The first cell line, which had an epithelial morphology and was established in DMEM (Dulbecco's modified Eagle's medium)/Ham's F-12 (DF)-based medium (RL/DF cells), expressed CK18 (cytokeratin 18), a marker of parenchymal hepatocytes. The other, a mesenchymal-like cell line established in DMEM-based medium (RL/DMEM cells), expressed αSMA (α-smooth muscle actin), a marker of hepatic myofibroblasts. Epithelial RL/DF cells underwent phenotypic changes, such as increased expression of αSMA, when the culture medium was switched to DMEM-based medium. In contrast, mesenchymal RL/DMEM cells were induced to express the epithelial marker CK18 with a concomitant decrease in αSMA expression when the culture medium was replaced with DF-based medium. These cell lines may provide novel in vitro models for the study of the conversion between epithelial and mesenchymal phenotypes during EMT in liver fibrosis and morphogenesis.

Abstract Image

SV40大T抗原永生化大鼠肝细胞系上皮和间充质表型的可逆转化
上皮间质转化(epithelial-mesenchymal transition, EMT)是肝纤维化的关键过程。这一过程也是胚胎发育过程中肝脏形态发生所必需的。为了研究EMT的细胞和分子基础,我们从大鼠肝细胞中建立了两种表型不同的SV40大T抗原永生化细胞系。在DMEM (Dulbecco's modified Eagle's medium)/Ham's F-12 (DF)培养基(RL/DF细胞)中建立的第一个具有上皮形态的细胞系表达CK18(细胞角蛋白18),这是实质肝细胞的标记物。另一个是在基于DMEM的培养基中建立的间充质样细胞系(RL/DMEM细胞),表达α-平滑肌肌动蛋白(α-平滑肌肌动蛋白),这是肝肌成纤维细胞的标志。将上皮细胞RL/DF转换为以dmem为基础的培养基后,细胞发生了αSMA表达增加等表型变化。相比之下,当用df培养基代替培养基时,诱导间充质RL/DMEM细胞表达上皮标记物CK18,同时αSMA表达降低。这些细胞系可能为研究EMT在肝纤维化和形态发生过程中上皮和间充质表型之间的转换提供新的体外模型。
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