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Murine hepatocellular carcinoma derived stem cells reveal epithelial-to-mesenchymal plasticity. | LitMetric

Murine hepatocellular carcinoma derived stem cells reveal epithelial-to-mesenchymal plasticity.

World J Stem Cells

the University of Queensland School of Medicine and the Gallipoli Medical Research Institute, Greenslopes Private Hospital, Brisbane, QLD 4120, Australia.

Published: September 2017

Aim: To establish a model to enrich and characterize stem-like cells from murine normal liver and hepatocellular carcinoma (HCC) cell lines and to further investigate stem-like cell association with epithelial-to-mesenchymal transition (EMT).

Methods: In this study, we utilized a stem cell conditioned serum-free medium to enrich stem-like cells from mouse HCC and normal liver cell lines, Hepa 1-6 and AML12, respectively. We isolated the 3-dimensional spheres and assessed their stemness characteristics by evaluating the RNA levels of stemness genes and a cell surface stem cell marker by quantitative reverse transcriptase-PCR (qRT-PCR). Next, we examined the relationship between stem cells and EMT using qRT-PCR.

Results: Three-dimensional spheres were enriched by culturing murine HCC and normal hepatocyte cell lines in stem cell conditioned serum-free medium supplemented with epidermal growth factor, basic fibroblast growth factor and heparin sulfate. The 3-dimensional spheres had enhanced stemness markers such as and and hepatic cancer stem cell (CSC) marker compared to parental cells grown as adherent cultures. We report that epithelial markers and were downregulated, while mesenchymal markers and were upregulated in 3-dimensional spheres. The 3-dimensional spheres also exhibited changes in expression of , and family of EMT transcription factors.

Conclusion: Our novel method successfully enriched stem-like cells which possessed an EMT phenotype. The isolation and characterization of murine hepatic CSCs could establish a precise target for the development of more effective therapies for HCC.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5620425PMC
http://dx.doi.org/10.4252/wjsc.v9.i9.159DOI Listing

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