Novel RNA oligonucleotide improves liver function and inhibits liver carcinogenesis in vivo.
Novel RNA oligonucleotide improves liver function and inhibits liver carcinogenesis in vivo.
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DOI：
10.1002/hep.26669
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发表时间：
2014-01
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影响因子：
--
通讯作者：
Habib NA
中科院分区：
文献类型：
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作者：
Reebye V;Sætrom P;Mintz PJ;Huang KW;Swiderski P;Peng L;Liu C;Liu X;Lindkaer-Jensen S;Zacharoulis D;Kostomitsopoulos N;Kasahara N;Nicholls JP;Jiao LR;Pai M;Spalding DR;Mizandari M;Chikovani T;Emara MM;Haoudi A;Tomalia DA;Rossi JJ;Habib NA
肝细胞癌（HCC）主要发生在肝硬化患者中。在这里，我们展示了一种基于 RNA 的创新靶向方法，可增强内源性白蛋白的产生，同时减少肝脏肿瘤负担。我们设计了短激活RNA（saRNA）来增强C/EBPα（CCAAT/增强子结合蛋白-α）的表达，C/EBPα是白蛋白基因表达的转录调节剂和激活剂。在 C/EBPα-saRNA 转染的 HepG2 细胞中，除了白蛋白分泌增加 3 倍和细胞增殖减少 50% 外，还观察到 C/EBPα 和白蛋白 mRNA 水平增加。在患有多灶性肝肿瘤的肝硬化大鼠模型中静脉注射 C/EBPα-saRNA 可使循环血清白蛋白增加 30% 以上，显示肝功能改善的证据。肿瘤负荷减少了 80% (p = 0.003)，肿瘤前转化标志物减少了 40%。由于 C/EBPα 已知通过视网膜母细胞瘤、p21 和细胞周期蛋白具有抗增殖活性；我们在 C/EBPα-saRNA 转染的 HepG2 细胞中使用 mRNA 表达肝癌特异性微阵列，以确认对细胞凋亡、血管生成和转移的负调控强烈富集的基因的下调。上调基因富集为肿瘤抑制因子和细胞分化的正调节因子。 C/EBPα-saRNA 转染细胞的定量 PCR 和蛋白质印迹分析表明，除了已知的 C/EBPα 抗增殖靶标外，我们还观察到 IL6R、c-Myc 的抑制和 STAT3 磷酸化的减少。我们首次证明，一种新型可注射 saRNA 寡核苷酸可增强 C/EBPα 表达，成功降低临床相关肝硬化/HCC 模型中的肿瘤负荷，同时改善肝功能。
Hepatocellular carcinoma (HCC) occurs predominantly in patients with liver cirrhosis. Here, we show an innovative RNA-based targeted approach to enhance endogenous albumin production whilst reducing liver tumour burden. We designed short-activating RNAs (saRNA) to enhance expression of C/EBPα (CCAAT/enhancer-binding protein-α), a transcriptional regulator and activator of albumin gene expression. Increased levels of both C/EBPα and albumin mRNA in addition to a 3-fold increase in albumin secretion and 50% decrease in cell proliferation was observed in C/EBPα-saRNA transfected HepG2 cells. Intravenous injection of C/EBPα-saRNA in a cirrhotic rat model with multifocal liver tumours increased circulating serum albumin by over 30% showing evidence of improved liver function. Tumour burden decreased by 80% (p = 0.003) with a 40% reduction in a marker of pre-neoplastic transformation. Since C/EBPα has known anti-proliferative activities via retinoblastoma, p21 and cyclins; we used mRNA expression liver cancer specific microarray in C/EBPα-saRNA transfected HepG2 cells to confirm down-regulation of genes strongly enriched for negative regulation of apoptosis, angiogenesis and metastasis. Up-regulated genes were enriched for tumour suppressors and positive regulators of cell differentiation. A quantitative PCR and Western-blot analysis of C/EBPα-saRNA transfected cells suggested that in addition to the known anti-proliferative targets of C/EBPα, we also observed suppression of IL6R, c-Myc and reduced STAT3 phosphorylation. We demonstrate for the first time that a novel injectable saRNA-oligonucleotide that enhances C/EBPα expression successfully reduces tumour burden and simultaneously improves liver function in a clinically relevant liver cirrhosis/HCC model.