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In vitro anti-motile effects of Rhoifolin, a flavonoid extracted from Callicarpa nudiflora on breast cancer cells via downregulating Podocalyxin-Ezrin interaction during Epithelial Mesenchymal Transition
- Source :
- Phytomedicine. 93:153486
- Publication Year :
- 2021
- Publisher :
- Elsevier BV, 2021.
-
Abstract
- Background : Callicarpa nudiflora (C. nudiflora), which is a medical herb in genus of Callicarpa, widely grows in the southern part of China. Several investigations had shown that this herb exerts anti-tumor effects. Ezrin is an important membrane-cytoskeleton-binding protein. By organizing membrane proteins and orchestrating their signal transduction, Ezrin contributes to modulation of cytoskeleton rearrangement in cell motility. Purpose To investigate the anti-motile properties of Rhoifolin (RFL), a flavonoid from C. nudiflora, and to determine whether its effects are related to the inhibition on Podocalyxin (PODXL)-Ezrin signal transduction. Methods : To determine suitable concentration of RFL and exposure time on breast cancer cells, the effects of RFL on viability of breast cancer cells were evaluated by MTT assay. Then, the anti-migratory properties of RFL were determined by AP 48 chamber system and ORISTM cell migration assay. F-actin in MDA-MB-231 cells was visualized by Alexa Fluor™ 488 conjugated Phalloidin. Immunoprecipitation was involved to access the effects of RFL on the interaction between Ezrin and PODXL. In addition, several EMT markers, including E-cadherin, Vimentin, Snail and Slug, were measured by Western Blotting assay and cell immunofluorescent analysis. Finally, the effects of RFL on cell migration, expression of Ezrin and EMT markers were verified by small interfering RNA (siRNA) mediated gene silencing. Results : We showed here that treatments with 10 and 40 μM of RFL induced significant inhibitions on cell migration and alterations on the location and organization of actin cytoskeleton in breast cancer cells. Next, it was found that RFL suppressed Ezrin phosphorylation and consequent interaction with PODXL, significantly. Also, this compound showed an obvious inhibitory effect on TGF-β1-induced EMT in MDA-MB-231 cells. Furthermore, data from RNA interfering assay confirmed that the inhibitory effects of RFL on Ezrin was enhanced by the deletion of Ezrin. Conclusion : RFL shows anti-motile properties on breast cancer cells, which is due to its potential to downregulate Podocalyxin-Ezrin interaction during Epithelial Mesenchymal Transition.
- Subjects :
- Small interfering RNA
Epithelial-Mesenchymal Transition
Sialoglycoproteins
Cell
Pharmaceutical Science
Breast Neoplasms
Vimentin
macromolecular substances
Callicarpa
Disaccharides
03 medical and health sciences
0302 clinical medicine
Ezrin
Cell Movement
Cell Line, Tumor
Drug Discovery
medicine
Humans
MTT assay
Glycosides
Epithelial–mesenchymal transition
030304 developmental biology
Flavonoids
Pharmacology
0303 health sciences
biology
Chemistry
Cell migration
Actin cytoskeleton
Cell biology
Cytoskeletal Proteins
medicine.anatomical_structure
Complementary and alternative medicine
030220 oncology & carcinogenesis
biology.protein
Molecular Medicine
Female
Subjects
Details
- ISSN :
- 09447113
- Volume :
- 93
- Database :
- OpenAIRE
- Journal :
- Phytomedicine
- Accession number :
- edsair.doi.dedup.....3e5cb3c01b8795ceae157752ff24f75b