Study On Effect Of Periplaneta Americana Extract On High Metastatic Ovarian Cancer Cell Line HO-8910PM

ObjectiveProbe into the effects of Periplaneta Americana extract on proliferation,apoptosis,cell migration and angiogenesis of human high metastatic ovarian cancer cell line HO-8910 PM and explore its relevant mechanism,to provide theoretical guidance for its clinical treatment of ovarian cancer.Methods1.The experimental group: Periplaneta americana extract(10,20,30,40mg/L)group and the blank group(without medicine).2.MTT method and cell scratch test were used to measure the influence of growth proliferation of different concentrations of Periplaneta Americana extract on HO-8910 PM.3.Western blot was used to detect the expression of AKT,p-AKT,P38,p-P38,ERK1/2,p-ERK1/2 and VEGF in HO-8910 PM under different concentrations of Periplaneta Americana to surmise the relationship between cell apoptosis and PI3K-AKT,Ras/Raf/MEK/ERK and P38 MAPK signaling pathways and the effects of Periplaneta Americana on HO-8910 PM angiogenesis.4.Using JC-1 dyeing to observe the changes of mitochondrial membrane potential of HO-8910 PM cells with different concentrations of Periplaneta Americana extract under a fluorescence microscope.5.The effects of Annexin V-FITC staining on cell proliferation and apoptosis of HO-8910 PM cells with different concentrations of Periplaneta Americana extract were measured by Flow Cytometry(FCM).6.Cell scratch test to detect the impact of Periplaneta Americana extract on cell migration.Results1.MTT method: Under same processing time,Periplaneta Americana extract can inhibit the proliferation of HO-8910 PM and with the increase of drug concentration its growth inhibition rate also increased,and calculated IC50 was(22.08±1.5)mg/L.2.Western blot: With the increase of Periplaneta Americana extract concentration,the phosphorylation levels of AKT,ERK1/2,P38 in cells gradually decreased,and the protein levels of AKT,P38 and ERK1/2 did not change significantly.With the concentration of Periplaneta Americana extract increased VEGF protein expression levels gradually decreased.3.JC-1 staining showed that compared with the blank group,the mitochondrial membrane potential of the drug group decreased more significantly.4.Flow cytometry showed that the apoptotic rates in the blank group and PA extract groups were(15.1 ± 0.21),(15.5 ± 0.47),(20.3 ± 0.89)and(27.1 ±0.70).The effect was concentration-dependent.When the concentration of PA was lower than 30 mg/L,the apoptosis rate increased with the increase of PA concentration.5.Cell scratch test: The cell proliferation ability of Periplaneta Americana drug group decreased significantly,and even large area cell death appeared with the increase of concentration.ConclusionsPeriplaneta Americana extract could inhibit the proliferation of high metastatic cell line HO-8910 PM,promote its cell apoptosis and inhibit its migration and angiogenesis,the mechanism of promoting apoptosis may be related to the decrease of p-AKT,p-ERK1/2 and p-P38 protein expression.