Objective  To investigate the regulation effect of Morusin on stemness phenotype of laryngeal cancer stem cells.

  Methods  Separation and detection the proportion of CD133⁺ laryngeal cancer stem cells through flow cytometry; evaluation the self-renewal ability of CD133⁺ laryngeal cancer stem cells by tumor sphere formation assay; exploring the migration ability of CD133⁺ laryngeal cancer stem cells by Transwell assay; analyzing the cytotoxicity of chemotherapy drugs on CD133⁺ laryngeal cancer stem cells by modified MTT assay; detection of the expression levels of stemness associated markers by immunofluorescence staining, RT-qPCR and Western blot. After treatment with different concentrations of Morusin, cells were performed the above experiments for detection the self-renewal ability, migration ability, cytotoxicity resistance and expression of stemness associated markers.

  Results  Flow cytometry analysis showed that the proportion of CD133⁺ laryngeal cancer stem cells was (3.50±0.34)%, while after enrichment, the proportion increased to (93.20±5.23)%. CD133⁺ laryngeal cancer stem cells exhibited better self-renewal ability (P<0.001) and migratory ability (P<0.05); they were resistant to the cytotoxicity of chemotherapy drug (P<0.05), and highly expressed of stemness associated markers. After being treated with Morusin, the self-renewal and migratory abilities of CD133⁺ laryngeal cancer stem cells were reduced (P<0.05). In addition, after treated with Morusin, CD133⁺ laryngeal cancer stem cells were more sensitive to chemotherapy drugs; moreover, the expression levels of stemness associated markers were decreased.

  Conclusion  CD133⁺ laryngeal cancer stem cells possessed stemness phenotypic characteristics. Morusin attenuated stemness phenotype of laryngeal cancer stem cells, which may be related to its down-regulation effect on stemness associated markers.