Elimination of Cancer Stem-Like “Side Population” Cells in Hepatoma Cell Lines by Chinese Herbal Mixture “Tien-Hsien Liquid”
Figure 2
Huh 7 side population cells possessed higher clonogenicity and expression of stem cell markers. (a) Cell sorting of Huh7 cells using Hoechst 33342. The R2 gate represented the SP cells (1.69% of total cells) and the R1 gate represented the non-SP cells. Both of them were collected for the subsequent experiments. (b) RT-PCR assay showed that the SP cells possessed higher expression of stem cell markers. (c) Clonogenic assay showing the clonogenicity of SP cells was markedly higher than that of non-SP cells. (d) Cell morphology of freshly sorted SP and non-SP cells at days 0, 3, and 9 after seeding. As shown in the photographs, the differences in the shapes of SP (small and round in shape) and non-SP cells (fibroblast-like) were remarkable. (e) Protein expression and nuclear location of -catenin in SP and non-SP cells. The SP cells had much higher nuclear -catenin than the non-SP cells. Similar phenomenon was also observed in the cytoplasmic -catenin level. Scale bar, 50 μm. Similar results of clonogenicity and stemness genes expression were also obtained in HepG2 and Hep3B SP cells (data not shown). NSP: non-SP cells.
