A Newly Established Murine Cell Line as a Model for Hepatocellular Cancer in Non-Alcoholic Steatohepatitis
A Newly Established Murine Cell Line as a Model for Hepatocellular Cancer in Non-Alcoholic Steatohepatitis
复制标题
DOI：
10.3390/ijms20225658
复制
发表时间：
2019-11-01
影响因子：
5.6
通讯作者：
Fragoulis, Athanassios
中科院分区：
文献类型：
--
作者：
Kroh, Andreas;Walter, Jeanette;Fragoulis, Athanassios
由于全球肥胖患病率的增加，非酒精性脂肪性肝炎（NASH）已成为肝细胞癌（HCC）的主要危险因素。然而，NASH及其向HCC发展的病理生理机制尚不完全清楚。因此，本研究的目的是产生一种模型特异性nash来源的HCC细胞系。我们建立了小鼠NASH-HCC模型，并研究了获得的癌细胞（N-HCC25）的染色体畸变、细胞类型特异性标志物的表达、对营养物质的依赖以及mTOR的功能重要性。与健康肝细胞相比，N-HCC25表现出一些染色体畸变。肝细胞（HNF4）、EMT （Twist、Snail）和癌症干细胞标志物（CD44、EpCAM、CK19、Sox9）在这些细胞中同时表达。增殖高度依赖于葡萄糖和FBS的供应，而不是谷氨酰胺。用第二代mTOR抑制剂（KU-0063794）治疗导致细胞生长以剂量依赖的方式强烈下降。相比之下，第一代mTOR抑制剂（依维莫司）仅能轻微降低细胞增殖。细胞周期分析显示，观察到的生长减少最有可能是由于G(1)/G(0)细胞周期阻滞。这些结果表明，N-HCC25是一种来自NASH背景的高增殖HCC细胞系，可能作为未来NASH源性HCC研究的合适体外模型。
Non-alcoholic steatohepatitis (NASH) has become a major risk factor for hepatocellular cancer (HCC) due to the worldwide increasing prevalence of obesity. However, the pathophysiology of NASH and its progression to HCC is incompletely understood. Thus, the aim of this study was to generate a model specific NASH-derived HCC cell line. A murine NASH-HCC model was conducted and the obtained cancer cells (N-HCC25) were investigated towards chromosomal aberrations, the expression of cell type-specific markers, dependency on nutrients, and functional importance of mTOR. N-HCC25 exhibited several chromosomal aberrations as compared to healthy hepatocytes. Hepatocytic (HNF4), EMT (Twist, Snail), and cancer stem cell markers (CD44, EpCAM, CK19, Sox9) were simultaneously expressed in these cells. Proliferation highly depended on the supply of glucose and FBS, but not glutamine. Treatment with a second generation mTOR inhibitor (KU-0063794) resulted in a strong decrease of cell growth in a dose-dependent manner. In contrast, a first generation mTOR inhibitor (Everolimus) only slightly reduced cell proliferation. Cell cycle analyses revealed that the observed growth reduction was most likely due to G(1)/G(0) cell cycle arrest. These results indicate that N-HCC25 is a highly proliferative HCC cell line from a NASH background, which might serve as a suitable in vitro model for future investigations of NASH-derived HCC.