The Ethylacetate Extract Of Blaps Rynchopetera Fairmaire Inhibited The Proliferation And Angiogenesis Of Ovarian Cancer

【Objective】To observe the effects of ethylacetate extract of Blaps rynchopetera Fairmaire on SKOV3 proliferation and angiogenesis of ovarian cancer,the study aims to clarity the mechanism of ethyl acetate extract of Blaps rynchopetera Fairmaire treat ovarian cancer.【Method】1.Extraction of ethylmethyl acetate from Blaps rynchopetera Fairmaire: The living adult Blaps rynchopetera Fairmaire was soaked in 60% ethanol,and the polar parts of ethyl acetate were extracted by petroleum ether,trichloromethane and ethyl acetate,and then the ethylacetate extract was obtained after lyophilization.2.cell experiment: The various concentrations ethylacetate extract of Blaps rynchopetera Fairmaire was divided into different concentrations(0μg/m L group,5μg/m L group,10μg/m L group,25μg/m L group,50μg/m L group,100μg/m L group),and cultured for 2h,6h,12 h,24h,36 h,48h,respectively.3.CCK-8 experiment: The optimal inhibitory concentration and inhibitory time of SKOV3 proliferation of ovarian cancer cells were determined by the ethylacetate extract of Blaps rynchopetera Fairmaire.4.Westernblot experiment: The expressions of β-catenin protein,Cyclin D1 protein and P21 protein were detected after different concentrations of ethylacetate extract of Blaps rynchopetera Fairmaire were treated for different time.5、Immunohistochemical experiment:The expression of VEGF and CD34 molecules were detected after the optimum treatment time of different concentrations of ethyl-acetate extract of Blaps rynchopetera Fairmaire【Results】1.The living adults were soaked in 60% ethanol for two rounds and then extracted and lyophilized by three organic solvents.Crude extracts of petroleum ether,trichloromethane and ethyl acetate from different polar parts were obtained successfully.Finally,ethylacetate extract was retained,and the yield of ethyl acetate was 0.2%.2.CCK-8 experiment: The optimal inhibitory concentration and the optimal inhibitory time of SKOV3 were 10μg/m L and 48 h.3.Westernblot experiment: SKOV3 cells were treated with ethylacetate extract of Blaps rynchopetera Fairmaire,the expression levels of β-catenin and Cyclin D1 were lower than0μg/m L at different concentrations for 48 h,and the expression levels of β-catenin and Cyclin D1 were the lowest at 10μg/m L.The expressions of β-catenin protein and Cyclin D1 protein decreased gradually with the increase of time when 10μg/m L was treated for different time,and the expressions of β-catenin protein and Cyclin D1 protein were the lowest at 48 h.The expression of P21 protein was higher than 0μg/m L at different concentrations for 48 h,and higher at 10μg/m L.When 10μg/m L was treated for different time,the expression of P21 protein decreased gradually with the increase of time,and the expression of P21 protein was the lowest at 48 hours.4.Immunohistochemical experiment: SKOV3 cells were treated with ethylacetate extract of Blaps rynchopetera Fairmaire.When treated with different concentrations for 48 h,the expression levels of VEGF and CD34 were decreased to different degrees,and the expression levels were the lowest at the concentration of 10μg/m L.When the concentration of 10μg/m L was treated for different time,the expression levels of VEGF and CD34 molecules gradually decreased with the increase of time.【Conclusions】1.The ethylacetate extract of Blaps rynchopetera Fairmaire has inhibition of tumor cell proliferation properties on ovarian cancer cells SKOV3 in vitro.2.The ethylacetate extract of Blaps rynchopetera Fairmaire down-regulated β-catenin and Cyclin D1 protein and up-regulated P21 protein on ovarian cancer cells SKOV3,could inhibit cell proliferation.3.The ethylacetate extract of Blaps rynchopetera Fairmaire down-regulated expression of VEGF and CD34 molecules on ovarian cancer cells SKOV3,could inhibit angiogenesis.