Inhibition of proliferation and migration of hepatocellular carcinoma-like stem cells and expression of stemness transcription factors by Plumbagin
Objective:To investigate the effects of plumbagin(PLB)on the proliferation,migration and expression of stem-ness transcription factors of hepatocellular carcinoma stem cells.Methods:The liver cancer stem cells were enriched by multi-cell tumor sphere culture mode.The liver cancer cell group and liver cancer stem cell group were set up with PLB concentration 0,5,7.5,10 μmol/L groups,and drug treatment time was 24 h.The cell proliferation,sphere formation and migration ability in vitro were detected by MTT assay,plate cloning assay,suspension stem cell sphere formation assay and scratch test.The expression of stemness genes and proteins was detected by qRT-PCR and Western blotting.Results:HepG2 stem cells were successfully en-riched.Compared with HepG2 cell group,the clone formation rate of HepG2 stem cells group was decreased,and the scratch heal-ing rate and sphere formation rate in vitro were increased,and the differences were statistically significant(P<0.05).Compared with the PLB 0 μmol/L group,the colony formation rate,sphere formation rate and scratch healing rate of HepG2 stem cells in the PLB 5,7.5,10 μmol/L groups were decreased.The mRNA expression levels of EpCAM,BMI1,c-MYC,Oct-4,Nanog stemness related genes and Notch signaling pathway were decreased,and the protein expression level of c-MYC was decreased(P<0.05).Conclusion:PLB can inhibit the proliferation,migration and sphere formation ability of hepatoma-like stem cells in vitro,thereby inhibiting the stemness of hepatoma cells.The mechanism may be related to the down-regulation of the mRNA expression levels of EpCAM,BMI1,c-MYC,Oct-4,Nanog stemness related genes and Notch signaling pathway,and the expression level of c-MYC protein.
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