Institute of Cellular Medicine, Newcastle University, Level 4 William Leech Building, Medical School, Framlington Place, Newcastle Upon Tyne NE2 4HH, UK.
Institute of Cellular Medicine, Newcastle University, Level 4 William Leech Building, Medical School, Framlington Place, Newcastle Upon Tyne NE2 4HH, UK.
Differentiation. 2018 Jul-Aug;102:10-18. doi: 10.1016/j.diff.2018.05.003. Epub 2018 May 22.
The rodent pancreatic AR42J-B13 (B-13) cell line differentiates into non-replicative hepatocyte-like cells in response to glucocorticoid mediated via the glucocorticoid receptor (GR). The aims of this study were to identify a human cell line that responds similarly and investigate the mechanisms underpinning any alteration in differentiation. Exposing the human pancreatic adenocarcinoma (HPAC) cell line to 1-10 µM concentrations of dexamethasone (DEX) resulted an inhibition of proliferation, suppressed carcinoembryonic antigen expression, limited expression of pancreatic acinar and hepatic gene expression and significant induction of the constitutively-expressed hepatic CYP3A5 mRNA transcript. These changes were associated with a pulse of genomic DNA methylation and suppressed notch signalling activity. HPAC cells expressed high levels of GR transcript in contrast to other nuclear receptors - such as the glucocorticoid-activated pregnane X receptor (PXR) - and GR transcriptional function was activated by DEX in HPAC cells. Expression of selected hepatocyte transcripts in response to DEX was blocked by co-treatment with the GR antagonist RU486. These data indicate that the HPAC response to glucocorticoid exposure includes an inhibition in proliferation, alterations in notch signalling and a limited change in the expression of genes associated with an acinar and hepatic phenotype. This is the first demonstration of a human cell responding to similarly to the rodent B-13 cell regarding formation of hepatocyte-like cells in response to glucocorticoid. Identifying and modulating the ablating factor(s) may enhance the hepatocyte-like forming capacity of HPAC cells after exposure to glucocorticoid and generate an unlimited in vitro supply of human hepatocytes for toxicology studies and a variety of clinical applications.
啮齿动物胰腺 AR42J-B13（B-13）细胞系在糖皮质激素受体（GR）介导的糖皮质激素作用下分化为非复制性肝样细胞。本研究旨在鉴定一种类似反应的人类细胞系，并研究分化过程中任何改变的机制。将人胰腺腺癌（HPAC）细胞系暴露于 1-10µM 浓度的地塞米松（DEX）中会导致增殖受到抑制、抑制癌胚抗原表达、限制胰腺腺泡和肝基因表达，并显著诱导固有表达的肝 CYP3A5 mRNA 转录物。这些变化与基因组 DNA 甲基化的脉冲和抑制 Notch 信号活性有关。HPAC 细胞表达高水平的 GR 转录物，与其他核受体（如糖皮质激素激活的孕烷 X 受体（PXR））形成对比，GR 转录功能在 HPAC 细胞中被 DEX 激活。DEX 对选定的肝细胞转录物的表达的影响被 GR 拮抗剂 RU486 的共同处理所阻断。这些数据表明，HPAC 对糖皮质激素暴露的反应包括增殖抑制、Notch 信号改变以及与腺泡和肝表型相关的基因表达的有限改变。这是首次证明人类细胞对糖皮质激素的反应类似于啮齿动物 B-13 细胞，在糖皮质激素作用下形成肝样细胞。鉴定和调节消融因子可能会增强 HPAC 细胞在暴露于糖皮质激素后的肝样细胞形成能力，并为毒理学研究和各种临床应用生成无限的人肝细胞体外供应。
