Open AccessInhibiting of self-renewal, migration and invasion of ovarian cancer stem cells by blocking TGF-β pathway
Author(s) -
Haiyan Wen,
Min Qian,
Jing He,
Meihui Li,
Qing Yu,
Zhengwei Leng
Publication year - 2020
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0230230
Subject(s) - epithelial–mesenchymal transition , vimentin , flow cytometry , cell migration , gentamicin protection assay , biology , cancer research , cancer stem cell , ovarian cancer , stem cell , smad , transforming growth factor , microbiology and biotechnology , in vitro , cancer , metastasis , immunology , biochemistry , genetics , immunohistochemistry
Objective To investigate the effect and mechanism of SB525334 on self-renewal, migration and invasion of ovarian cancer stem cells. Methods ALDHhigh-expressing cancer stem cells (CSCs) were isolated from human ovarian cancer cell line SKOV-3 by flow cytometry and treated with 2μg/mL SB525334 for 6h. The sphere forming assay was used to detect the ability of self-renewal of CSCs and the colony formation assay was used to detect the tumorigenicity in vitro . Transwell migration and invasion assay were used to detect the migration and invasion ability of CSCs. To further explore the mechanism, real-time quantitative PCR and flow cytometry were used to detect the mRNA and protein expression of TGF-β, Smad2, Smad3, phosphorylated Smad2, phosphorylated Smad3 and Smad4, respectively. Expressions of epithelial-mesenchymal transition (EMT)-related genes E-cadherin, Snail, Vimentin were also assessed. Results The self-renewal ability, tumorigenicity in vitro , migration and invasion ability of CSCs were significantly attenuated after SB525334 treatment. The expressions of TGF-β, phosphorylated Smad2, phosphorylated Smad3, Snail, and Vimentin were decreased, while Smad4 and E-cadherin expressions were increased. Conclusion SB525334 may inhibit the self-renewal, invasion and migration of ovarian CSCs by blocking the TGF-β/Smad/EMT pathway.