Cinobufotalin as a Novel Agent to Inhibit in-Vitro Epithelial Ovarian Cancer Cell Proliferationc, Migrat Ion and Invasion

Abstract

ObjectiveCinobufotalin (CINO), a cardiotonic steroid (CTS) or bufadienolide, is extracted from the skin secretions of giant toads and is utilized in traditional Chinese medicine (Chan Su). CINO has been used as a cardiotonic, diuretic and a hemostatic agent. Our lab is familiar with CINO and has shown it to inhibit cytotrophoblast cells function. Recently, it has been shown that CINO also inhibits the lung cancer cell function, and has been further implicated in other disease processes. In the present study, we propose to pursue this potential application of CINO using ovarian tumor cell line SK-OV-3.Study DesignWe evaluated the in-vitro effect of CINO on ovarian cancer cell line SK-OV-3. Cells were treated with 0.1, 1, 5, and 10 µM CINO. Cell proliferation was measured using a CellTiter Assay (Promega), which is a colorimetric method for determining the number of viable cells. Cell migration was measured using a CytoSelect Assay (Cell Biolabs). Cell invasion was measured using a FluoroBlok Assay (BD). Cell viability was measure using a CellTiter Assay (Promega). Cell cycle progression was evaluated by a Cell Cycle Phase Determination Kit (Cayman Chemical) and apoptosis was evaluated by an Apoptotic Blebs Assay Kit (Cayman Chemical). Cell cycle arrest and apoptotic signaling was determined by fluorescence-activated cell sorting (FACS) analysis.ResultsCINO at ≥0.5 µM inhibited SKOV-3 cell proliferation, migration, and invasion (p<0.05). There was a higher (p<0.05) percentage of S phase cells in groups treated with CINO at 0.5 µM. CINO at ≥0.5 µM down regulated expression of PCNA and caused cell death.ConclusionThis data demonstrates that CINO impairs SK-OV-3 cell function via cell cycle arrest and apoptotic signaling. These findings demonstrate the complex nature of this compound. Not only is CINO directly modulating the actions of the Na/K ATPase through classic mechanism of cardiotonic steroids, but is also directly influencing the nuclear expression of proteins involved in cell cycle progression and DNA repair. Additional investigational studies looking into the molecular pathways involved in altering cell cycle and entry into apoptosis are warranted.In conclusion, we have shown CINO to impair SK-OV3 cell function via cell cycle arrest and apoptotic signaling and suggest that CINO might be further investigated as a novel anti-ovarian cancer agent.

Differential Effects of In Vitro Treatment with Cinobufotalin on Three Types of Ovarian Cancer Cells

“…According to our study, reduced expression of PCNA was observed across all three cell lines on treatment with cinobufotalin. In a prior study by McDowell et al with SK-OV-3 ovarian cancer cells, it was observed that cinobufotalin concentrations exceeding 0.5 μM reduced PCNA expression and cell viability and induced apoptotic signaling (19). CRL-1978 and CRL-11731 lines exhibited similar trends.…”

“…Morphologically it is an epithelial cell line from a French-Canadian ancestry patient with early onset ovarian cancer. A recent study from our laboratory demonstrated that cinobufotalin at in vitro concentrations greater than 0.5 μM inhibited SK-OV-3 cell proliferation, migration and invasion (19). The goal of this study was to determine if the cytotoxic effect of cinobufotalin is cell-specific or if it affects each cancer cell line similarly.…”

Differential Mechanism of Action of 3,4’,7-O-trimethylquercetin in Three Types of Ovarian Cancer Cells

“…Studies are being constantly performed to understand the effect of different novel compounds on proliferation and migration of cells (14,15). McDowell et al examined the effect of a novel compound, cinobufotalin (CINO), on an ovarian cancer cell line in vitro and found that it decreased the proliferation, migration, and invasion of the cancer cells (16). It was also found that CINO was able to hinder the progression of cytotrophoblast cells and ovarian cancer cells by inducing cell cycle arrest and apoptosis (16,17).…”

“…McDowell et al examined the effect of a novel compound, cinobufotalin (CINO), on an ovarian cancer cell line in vitro and found that it decreased the proliferation, migration, and invasion of the cancer cells (16). It was also found that CINO was able to hinder the progression of cytotrophoblast cells and ovarian cancer cells by inducing cell cycle arrest and apoptosis (16,17). Studies like these opened an avenue to discover and explore the benefit of other natural compounds such as Quercetin.…”

MiR-145 on the Proliferation of Ovarian Cancer Cells by Regulating the Expression of MMP-2/MMP-9

“…In order to further explore its effect on the biological behavior of OC cells and its related molecular mechanisms, people constructed a pCMV-N-Flag-PinX1 overexpression recombinant vector, which was transiently transfected into human OC cell SKOV3 by liposome, and The expression changes of PinX1 in cells were detected at the mRNA and protein levels, and the results showed that the expression level of PinX1 was significantly up-regulated and the PinX1 gene could be successfully expressed. However, the successful construction of a stable over-expressed PinX1 vector will likely become a gene target A powerful tool for treatment (30,31). In addition, CCK8, research results show that after overexpression of PinX1, SKOV3 cell viability is inhibited, and cell invasion ability is significantly reduced, suggesting that in OC cells, low-expression PinX1 may promote the malignant proliferation of OC, and infiltration and metastasis of tumors closely related (32,33).…”