Effects Of Casticin On Production Of Reactive Oxygen Species And Apoptosis In Ovarian Cancer HO-8910Cells

ObjectiveTo observation of reactive oxygen species（ROS） in casticin-induced of ovariancancer HO-8910cells apoptosis, and explore induction of apoptosis by CAS isinvolved in ROS-mediated mitochondrial apoptosis signaling pathway.MethodsHuman ovarian cancer HO-8910cells were cultured in vitro. The apoptotic rate wasanalyzed by flow cytometry (FCM) with Annexin V-FITC/PI double staining.Intracellular ROS was measured by FCM with fluorescent probe2’,7’-dichlorodihydrofluorescein diacetate (H2DCFH-DA). The FCM with rhodamine123was used to analyze the mitochondrial transmembrane potential. Western Blottingwas used to analyze the expression of Cyt C, Bax and Bcl-2protein in HO-8910cells.ResultFCM analysis with Annexin V-FITC/PI double staining indicated that the apoptoticrate by treatment with2.0,4.0μg/mL CAS for48h was9.46±0.95%and14.47±1.47%,respectively, and the difference has statistical significance comparison with theNAC group(4.63±0.57%，5.30±0.50%)and the vehicle group(0.2%DMSO，0.3±0.10%).FCM analysis with H2DCFH-DA staining showed that the Mean fluorescenceintensity by treatment with2.0,4.0μg/mL CAS for48h was36.73±1.52，57.53±2.83，and the difference has statistical significance comparison with the NACgroup(13.63±1.25，14.63±1.60)..FCM analysis with rhodamine123stainingindicated that the mitochondrial transmembrane potential by treatment with 2.0,4.0μg/mL CAS for48h was12.73±1.25、18.20±1.49，and apparently lower thanthe NAC group(56.20±2.73，59.7±1.79)and the vehicle group(0.2%DMSO,84.67±3.73).. Western blotting showed that HO-8910cell treated with CAS(2.0and4.0μg/mL) for48h upregulated the expression of CytC, Bax protein, the averagerelative gray scale values were1.604±0.327,1.781±0.21(P <0.05) and1.311±0.325,1.665±0.030(P <0.05), compared to solvent control group (1.258±0.456,1.047±0.356) a statistically significant difference (P <0.05).The expression ofBcl-2protein decreased,average relative gray value of1.361±0.0241.302±0.010(P <0.05), the vehicle control group was1.674±0.217.ConclusionsCAS-induced apoptosis of human ovarian cancer HO-8910cells may be associatedwith ROS-mediated mitochondrial apoptosis signaling pathway.