Tumour stroma and microenvironment have been shown to affect hepatocellular carcinoma (HCC) growth, with activated hepatic stellate cells (HSC) as a major contributor in this process. Recent evidence suggests that the energy sensor adenosine monophosphate-activated kinase (AMPK) may mediate a series of essential processes during carcinogenesis and HCC progression. Here, we investigated the effect of different HCC cell lines with known TP53 or CTNBB1 mutations on primary human HSC activation, proliferation and AMPK activation. We show that conditioned media obtained from multiple HCC cell lines differently modulate human hHSC proliferation and hHSC AMPK activity in a paracrine manner. Pharmacological treatment of hHSC with AICAR and Compound C inhibited the HCC-induced proliferation/activation of hHSC through AMPK-dependent and AMPK-independent mechanisms, which was further confirmed using mouse embryonic fibroblasts (MEFs) deficient of both catalytic AMPKα isoforms (AMPK^α1/α2-/-) and wild type (wt) MEF. Both compounds induced S-phase cell-cycle arrest and, in addition, AICAR inhibited the mTORC1 pathway by inhibiting phosphorylation of 4E-BP1 and S6 in hHSC and wt MEF. Datamining of the Cancer Genome Atlas (TCGA) and the Liver Cancer (LICA-FR) showed that AMPKα1 (PRKAA1) and AMPKα2 (PRKAA2) expression differed depending on the mutation (TP53 or CTNNB1), tumour grading and G1-G6 classification, reflecting the heterogeneity in human HCC. Overall, we provide evidence that AMPK modulating pharmacological agents negatively modulate HCC-induced hHSC activation and may therefore provide a novel approach to target the mutual, tumour-promoting interactions between hHSC and HCC.

中文翻译：

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AICAR和化合物C在体外负调控HCC诱导的人类肝星状细胞的活化

肿瘤基质和微环境已显示会影响肝细胞癌（HCC）的生长，其中活化的肝星状细胞（HSC）是此过程的主要贡献者。最近的证据表明，能量传感器单磷酸腺苷激活激酶（AMPK）可能介导一系列在癌变和HCC进程中的基本过程。在这里，我们研究了具有已知TP53或CTNBB1突变的不同HCC细胞系对人原代HSC激活，增殖和AMPK激活的影响。我们显示，从多个HCC细胞系获得的条件培养基以旁分泌方式不同地调节人hHSC增殖和hHSC AMPK活性。^{α1/α2 -/-}）和野生型（wt）MEF。两种化合物均诱导S期细胞周期停滞，此外，AICAR通过抑制hHSC和wt MEF中4E-BP1和S6的磷酸化来抑制mTORC1途径。癌症基因组图谱（TCGA）和肝癌（LICA-FR）的数据挖掘表明AMPKα1（PRKAA1）和AMPKα2（PRKAA2）的表达因突变（TP53或CTNNB1），肿瘤分级和G1-G6分类而异人类肝癌的异质性。总体而言，我们提供的证据表明，AMPK调节药理学药物可负面调节HCC诱导的hHSC活化，因此可能提供一种靶向hHSC与HCC之间相互促进肿瘤相互作用的新颖方法。