Angiolini Virginia Andrea, Cruz Carolina Uribe, López Mónica Luján, Simon Laura, Matte Ursula
Gene Therapy Center, Hospital de Clínicas de Porto Alegre, Rua Ramiro Barcelos 2350, Porto Alegre, RS 90035-903 Brazil ; Post-Graduation Program in Child and Adolescent Health, Federal University of Rio Grande do Sul, Porto Alegre, Brazil.
Gene Therapy Center, Hospital de Clínicas de Porto Alegre, Rua Ramiro Barcelos 2350, Porto Alegre, RS 90035-903 Brazil.
Cancer Cell Int. 2017 Jan 4;17:5. doi: 10.1186/s12935-016-0370-x. eCollection 2017.
Hepatocellular carcinoma is a common cancer, ranking third in cancer-associated deaths. An important cause of cancer patients' mortality is metastasis. At the start of metastasis progression, there is an epithelial-mesenchymal transition, characterized by matrix degradation, junction reductions and vessels formation. HuH-7 is a cell line used in research as an in vitro model for hepatocellular carcinoma. It is known that two-dimensional growth reflects tumor characteristics poorly. In contrast, three-dimensional cultures provide a better approach to the study of tumorigenic potential. The purpose of this work was to mimic a three-dimensional environment in order to assess gene expression of some epithelial-mesenchymal transition and metastasis progression markers in HuH-7 cells and compare them with traditional two-dimensional culture model.
HuH-7 cells were encapsulated in sodium alginate (three-dimensional model) to be compared with cells grown in two-dimensional flasks. After 4 days in culture, gene expression of , , and was analyzed by PCR and cytoskeleton assessment was performed by rhodamine-phalloidin staining.
Differences were found in gene expression, with a high increment of and reduction. The cytoskeleton morphology also showed differences, with a cytoplasm restricted only near the nuclei in the three-dimensional model.
This work shows the effects of using sodium alginate capsules as a three-dimensional model to the study of HuH-7. Cells in this 3D system show key markers of epithelial-mesenchymal transition, such as overexpression and down-regulation.
肝细胞癌是一种常见癌症，在癌症相关死亡中排名第三。癌症患者死亡的一个重要原因是转移。在转移进展开始时，会发生上皮-间质转化，其特征为基质降解、连接减少和血管形成。HuH-7是一种用于研究的细胞系，作为肝细胞癌的体外模型。已知二维生长很难反映肿瘤特征。相比之下，三维培养为研究致瘤潜力提供了更好的方法。这项工作的目的是模拟三维环境，以评估HuH-7细胞中一些上皮-间质转化和转移进展标志物的基因表达，并将其与传统二维培养模型进行比较。
将HuH-7细胞封装在海藻酸钠中（三维模型），与在二维培养瓶中生长的细胞进行比较。培养4天后，通过PCR分析、、和的基因表达，并通过罗丹明-鬼笔环肽染色进行细胞骨架评估。
发现基因表达存在差异，和的表达有高度增加，而减少。细胞骨架形态也显示出差异，在三维模型中，细胞质仅局限于细胞核附近。
这项工作展示了使用海藻酸钠胶囊作为三维模型对HuH-7进行研究的效果。该三维系统中的细胞显示出上皮-间质转化的关键标志物，如的过表达和的下调。
