Cancer stem cells (CSCs) are believed to be responsible for
tumor metastasis, recurrence, and high mortality of
cancer patients due to their high tumorigenicity resistance to chemo-
radiotherapy.
Morusin possesses anti-
cancer activity through attenuation of NF-κB activity, which is up-regulated in cancer stem cells. The purpose of this study is to confirm the growth and migration inhibition effect of
morusin on human cervical CSCs, and to clarify its partial mechanism of activity. Human cervical CSCs were enriched using non-adhesive culture system. Their stemness characteristics were identified with
tumor sphere formation, self-renewal,
toluidine blue staining, migration assays, RT-PCR analysis, and immunofluorescence staining of putative stem cell markers, Oct4, SOX2, and ALDH1; the epithelial-to-mesenchymal (EMT) transition markers and relevant
transcription factors were evaluated with Western blotting. The growth and migration inhibition effects of
morusin on human cervical CSCs were tested by cell proliferation,
tumor sphere formation, and transwell assay; apoptotic death of human cervical CSCs in response to
morusin was measured with
DAPI staining, apoptotic DNA fragmentation; NF-κBp65, Bcl-2, Bax, and
caspase-3 protein expressions were detected through Western blotting. Under this non-adhesive culture system, typical
tumor spheres appeared within 5-7 days, the
tumor sphere formation, self-renewal, and cell migration, expressions of putative stem cell markers, EMT markers, and relevant
transcription factors of the
tumor sphere cells were increased significantly. After
morusin treatment, the proliferation,
tumor sphere formation, and migration of human cervical CSCs were decreased significantly,
DAPI-stained apoptotic cells increased, apoptotic
DNA fragmentations formed evidently; the expression levels of NF-κBp65 and Bcl-2 decreased significantly, Bax, and
caspase-3 increased significantly in a dose-dependent manner. Using the non-adhesive culture system, human cervical CSCs were enriched and expanded.
Morusin has the potential to target and kill CSCs, and can inhibit human cervical growth and migration through NF-κB attenuation mediated apoptosis induction.