ABSTRACT
Background and purpose:Ovarian cancer is associated with a high recurrence and mortality due to the existence of cancer stem cells. The purpose of this study was to investigate the effect of casticin (CAS) on the capability of self-renewal in ovarian cancer stem cell like cells (OCSLCs) derived from SKOV3 cell line. Methods:SKOV3 cell line cells were cultured in vitro, and OCSLCs were obtained and amplified through suspended culture with conditioned medium of the stem cells. The phosphorylation level of FoxO3a was analyzed using Western blot. The protein expression of FoxO3a was inhibited by FoxO3a speciifc siRNA transfection, and then the ratio of sphere-formation was detected. Results: Compared with parental cells, OCSLCs over-expressed phosphorylated FoxO3a (pFoxO3a) and had elevated ratio of sphere-formation [(3.1±0.3)% vs (34.8±6.8)%, P<0.05]. CAS significantly inhibited the capability of sphere-formation in OCSLCs and down-regulated the expression level of pFoxO3a. And the transfection of FoxO3a speciifc siRNA suppressed the protein expression of FoxO3a and attenuated the inhibitory effect of CAS on the sphere-formation of OCSLCs. Conclusion: Reduced expression level of pFoxO3a is involved in the effect that CAS inhibits sphere-formation of OCSLCs derived from SKOV3 cell line.
ABSTRACT
Objective To investigate whether 5, 7-dihydrox-8-nitrochrysin (NOC) induces apoptosis in U937 monocytic leukae-mia cells is involved in the regulation of the activity of PKD 2.Methods U937 cell line cells were cultured in vitro .Apoptosis rate was analyzed by flow cytometry (FCM) using propidium iodide (PI) staining.DNA ladder bands were observed by DNA agarose gel electro-phoresis.The phosphorylated protein expression of PKD 2 was analyzed using Western blot .Results NOC (2.5, 5.0, and 10.0μmol/L) increased apoptosis rate in U937 cells in a concentration-dependent manner ( P <0.05).After treatment with NOC (5.0 and 10.0μmol/L) for 24 h, U937 cells presented typical DNA ladder bands .At the same time, not only did NOC effectively down-regulate the ex-pression of PDK2 phosphorylated protein , but also increased apoptosis rate in U 937 cells in the presence of G?6976, a specific inhibitor of PKD2.Conclusions The effect that NOC induces apoptosis in U 937 cells is related to the inhibition of the activity of PKD 2.
ABSTRACT
Aim To investigate whether 8-bromo-7-methoxychrysin ( BrMC ) can reverse the epithelial-mesenchymal transition ( EMT) in sphere-forming cells (SFCs) derived from cervical cancer SiHa cell line. Methods Human cervical cancer SiHa cell line was cultured in vitro and then SFCs were obtained from Si-Ha cell line using suspension culture with the stem-condition culture system. After treatment with BrMC for 72 h, the morphology of SFCs of SiHa cell line was observed via monolayer adherence culture. The expres-sion levels of E-cadherin, N-cadherin and Twist1 pro-teins were analyzed using western blot assay. Results SFCs of SiHa cell line presented the mesenchyme-like( spindle-shaped) cell shape and exhibited reduced E-cadherin and elevated N-cadherin protein expres-sion, compared with the parental cells. After treatment with BrMC(1. 0 μmol·L-1 ) for 72 h, SFCs of SiHa cell line changed from spindle-shaped cells in to poly-gon shaped cells under monolayer adherence culture. After treatment with BrMC for 24 h, the expression lev-els of E-cadherin protein were up-regulated and the ex-pression levels of N-cadherin and Twist1 proteins were decreased in SFCs derived from SiHa cell line. Con-clusion BrMC effectively reverses EMT of SFCs de-rived from cervical cancer SiHa cell line, and its mechanism is involved in down-regulating the expres-sion of Twist protein.