TGF‐β‐mediated LEFTY/Akt/GSK‐3β/Snail axis modulates epithelial‐mesenchymal transition and cancer stem cell properties in ovarian clear cell carcinomas

Abstract

Advanced ovarian clear cell carcinoma (OCCCa) shows poor prognosis with chemoresistance, which is associated with epithelial-mesenchymal transition (EMT)/cancer stem cell (CSC) features. The left-right determination factor (LEFTY), a novel member of the TGF-β superfamily, is a marker of stemness. Here we focused on the functional roles of LEFTY in OCCCas. OCCCa cell lines that were cultured in STK2, a serum-free medium for mesenchymal stem cells, or treated with TGF-β1 underwent morphological changes toward an EMT appearance, along with increased expression of LEFTY and Snail. The cells also showed CSC properties, as demonstrated by increases in the aldehyde dehydrogenase (ALDH)1 activity population, number of spheroid formation, and expression of several CSC markers. Inhibition of LEFTY expression induced decreases in the number of spindle-shaped cells and CSC features, while cells stably overexpressing LEFTY exhibited enhancement of such EMT/CSC properties. Finally, treatment of cells with TGF-β1 led to increased LEFTY expression and activation of Akt, which subsequently induced inactivation of GSK-3β, while inhibition of GSK-3β resulted in increased expression of both LEFTY and Snail. In clinical samples, LEFTY expression showed a tendency for positive associations with expression of vimentin, as well as Sox2 and ALDH1, in OCCCas with epithelial-like morphology, indicating a possible relationship between LEFTY and the epithelial-mesenchymal hybrid stage of the tumors. In conclusion, TGF-β-mediated LEFTY/Akt/GSK-3β/Snail axis may contribute to the establishment and maintenance of phenotypic characteristics of OCCCas through modulation of EMT/CSC properties.

Polyphenols Extracted from Artemisia annua L. Exhibit Anti-Cancer Effects on Radio-Resistant MDA-MB-231 Human Breast Cancer Cells by Suppressing Stem Cell Phenotype, β-Catenin, and MMP-9

“…The third one would be about whether β-catenin and MMP-9 are involved in RT resistance. To clearly answer this question, further study is needed, but β-catenin and MMP-9 are important molecules clearly involved in EMT [22][23][24]34] and cancer progression [25,26,35]. In addition, these proteins were significantly increased in RT-R MDA-MB-231 cells.…”

“…WNT/beta-catenin mediates the radiation resistance of breast progenitor cells [14]. β-catenin is an important molecule involved in EMT [22][23][24]. MMP-9 is most widely associated with cancer progression, due to its role in extracellular matrix remodeling and angiogenesis [25,26].…”

NT5E inhibition suppresses the growth of sunitinib‐resistant cells and EMT course and AKT/GSK‐3β signaling pathway in renal cell cancer

“…Moreover, Treesuwan et al found Cypripedin diminished an epithelial-to-mesenchymal transition through suppression of Akt/GSK-3β signaling in non-small cell lung cancer cells (32). In the study by Matsumoto et al, Transforming growth factor (TGF)-β-mediated LEFTY/Akt/GSK-3β/Snail axis modulates EMT and cancer stem cell properties in ovarian clear cell carcinomas (33). In order to confirm the effect of NT5E expression on EMT and AKT/GSK-3β signaling pathway in our study, EMT and AKT/GSK-3β signaling pathway-associated proteins were examined by Western blot.…”

Upregulation of fibronectin following loss of p53 function is a poor prognostic factor in ovarian carcinoma with a unique immunophenotype

“…(Supplementary Figure S1), as described previously [13,17]. In addition, spindle-shaped cells were defined as those that showed narrow and elongated phenotypes, along with weak or absent adhesions between cells, as described previously [17].…”

“…Four OCCCa cell lines, OVISE, ES2, OVTOKO, and TOV-21G were used as described previously [13,16,17], and two OHGSeCa cell lines, OVSAHO and OVCAR-3, were obtained from the National Institute of Biomedical Innovation (Osaka, Japan) and the American Type Culture Collection (Manassas, VA, USA), respectively. p53 shRNA knockdown cells were established using OVISE cells, which have a wild-type p53 gene and abundant expression of endogenous HNF-1b and ARID1A…”