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Abstract

Purpose: The side population (SP) is thought enriched with hepatocellular cancer stem cells (HCSCs); however, it?s properties has not been fully explored in fresh tissues of hepatocellular cancer (HCC). Methods: Hoechst33342 was used to isolate the SP cells from HCC cell line HepG2 and fresh tumor tissues of 21 HCC patients by flow cytometry. Properties of the SP cells were established by cell proliferation assay, capacity to form spheroids in the serum-free medium, invasion and migration assays, tumorigenicity in NOD/SCID mice and mRNA microarray analysis. Results: SP cells were observed in HepG2 and 18 of the 21 fresh tissue samples. SP cells showed higher abilities of forming spheroids, invasion and migration, and there was no significant difference in cells proliferation between the two groups. Tumors could generate from SP but not non-SP (NSP) cells in a low dose of subcutaneous injection to the NOD/SCID mice (5×102cells/ mouse). The mRNA microarray analysis in HepG2 SP cells revealed that 2057 gene expressions were up-regulated and 3189 down-regulated. Conclusion: Our work indicates that SP cells can be sorted from HepG2 and HCC fresh samples. Compared with NSP cells, SP has the properties of higher invasion, migration and stemness.

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