Molecular mechanisms regulating the establishment of hepatocyte polarity during human hepatic progenitor cell differentiation into a functional hepatocyte-like phenotype
Molecular mechanisms regulating the establishment of hepatocyte polarity during human hepatic progenitor cell differentiation into a functional hepatocyte-like phenotype
复制标题
在人肝祖细胞分化为功能性肝细胞样表型过程中调节肝细胞极性建立的分子机制。
DOI：
10.1242/jcs.110551
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发表时间：
2012-12-01
影响因子：
4
通讯作者：
Zhang, Haiyan
中科院分区：
文献类型：
--
作者：
Hua, Mingxi;Zhang, Weitao;Zhang, Haiyan
肝细胞的正常功能需要肝细胞极性的建立和维持。然而，调节肝细胞极性产生的机制还不完全清楚。人胎肝祖细胞（hFHPCs）向功能性肝细胞的分化为研究肝细胞发育的分子机制提供了一个强有力的体外模型系统。在这项研究中，我们使用了两个阶段的分化协议，以产生功能极化肝细胞样细胞（HLC）从hFHPC。对hFHPC、不成熟-HLC和成熟-HLC的一式三份样品进行全局基因表达谱分析。当基于分化阶段比较差异基因表达时，鉴定了一些可能对建立和维持肝细胞极性至关重要的基因。这些基因包括编码肌动蛋白抑制剂结合蛋白、蛋白酪氨酸激酶活性分子和信号传导途径组分的基因，如PTK 7、PARD 3、PRKCI和CDC 42。基于已知和预测的蛋白质-蛋白质相互作用，候选基因被分配到网络中并聚类到功能类别中。使用实时RT-PCR证实了这些基因中的几个的表达。通过使用小干扰RNA使基因失活，我们证明了PTK 7和PARD 3促进肝极性形成并影响F-肌动蛋白组织。这些结果提供了独特的见解，在肝细胞分化过程中的极化复杂的过程中，指示关键基因和信号分子控制肝细胞分化。
The correct functioning of hepatocytes requires the establishment and maintenance of hepatocyte polarity. However, the mechanisms regulating the generation of hepatocyte polarity are not completely understood. The differentiation of human fetal hepatic progenitor cells (hFHPCs) into functional hepatocytes provides a powerful in vitro model system for studying the molecular mechanisms governing hepatocyte development. In this study, we used a two-stage differentiation protocol to generate functional polarized hepatocyte-like cells (HLCs) from hFHPCs. Global gene expression profiling was performed on triplicate samples of hFHPCs, immature-HLCs and mature-HLCs. When the differential gene expression was compared based on the differentiation stage, a number of genes were identified that might be essential for establishing and maintaining hepatocyte polarity. These genes include those that encode actin filament-binding protein, protein tyrosine kinase activity molecules, and components of signaling pathways, such as PTK7, PARD3, PRKCI and CDC42. Based on known and predicted protein-protein interactions, the candidate genes were assigned to networks and clustered into functional categories. The expression of several of these genes was confirmed using real-time RT-PCR. By inactivating genes using small interfering RNA, we demonstrated that PTK7 and PARD3 promote hepatic polarity formation and affect F-actin organization. These results provide unique insight into the complex process of polarization during hepatocyte differentiation, indicating key genes and signaling molecules governing hepatocyte differentiation.