Experimental Study On The Anti-ovarian Cancer Effect Of Oleuropein In Vitro And In Vivo

Objective:To observe the effects of different concentrations of oleuropein and different action time on the proliferation of ovarian cancer SKOV3 cells,and the effects of oleuropein on the growth of transplanted tumors of nude mice subcutaneous SKOV3 cells,and to explore the possible mechanism of oleuropein against ovarian cancer.Methods:Ovarian cancer SKOV3 cells were cultured and inoculated on five 96-well plates.Four of them were experimental groups respectively added with oleuropein at a concentration of 100,200,400 and 800 ?g/ml,and the rest was set up as a control group(no oleuropein intervention).The proliferation inhibition of SKOV3 cells with different drug concentrations at three time points of 24 H,48H and 72 H was detected by CCK-8 assay.A model of subcutaneous tumor bearing SKOV3 cells in the back of nude mice was established.Twenty nude mice bearing tumors were randomly divided into two groups: oleuropein group: 50 mg·10ml·kg-1 intraperitoneally,saline group: 0.9 % sodium chloride injection 10 ml · kg-1 intraperitoneal injection.Both groups were administered once every other day for 8 weeks,free water intake,and standard SPF feed.The tumor volume and body weight of nude mice were measured and recorded once a week.After the nude mice were sacrificed,the tumor tissues were completely isolated and the relative proliferation rate of the tumors was calculated.Western blot was used to detect the expression levels of pro-apoptosis factor Bax and anti-apoptosis factor Bcl-2 in the two groups of tumor tissues.Results : In vitro experiments,compared with the control group,oleuropein concentration of 100,200,400,800ug/ml inhibited the proliferation of SKOV3 cells,the difference was statistically significant(P<0.05);At different time points,The oleuropein at 800ug/ml had the strongest inhibitory effect on cells(P<0.05).In the drug concentration of 800ug/ml,the cells grew slowly.There was no statistical difference in cell proliferation at the two time points of 24 h and 48H(p>0.05),but compared with 24 h and 72 h,cell proliferation was statistically significant(p<0.05);at the same concentration,with the prolongation of the time of drug action,there was no significant difference in inhibition rate(P> 0.05).In vivo,the tumor volume of oleuropein group in nude mice was significantly smaller than that of saline group(P<0.05);the relative tumor proliferation rate of oleuropein group was 70.3%;compared with saline group,oleuropein group Bax protein expression was up-regulated while Bcl-2 protein expression was down-regulated.Conclusion:Oleuropein can inhibit the proliferation of ovarian cancer SKOV3 cells in vitro and inhibit the growth of ovarian cancer SKOV3 cells subcutaneous transplanted tumor in nude mice.It can play an anti-ovarian cancer role by inducing apoptosis and inhibiting cell proliferation.The mechanism may be related to the enhancement of the expression of apoptosis-promoting factor Bax,the inhibition of the expression of anti-apoptosis factor Bcl-2,and may be related to the regulation of proto-oncogenes or tumor suppressor genes in cancer cells.The specific mechanism needs further study,and oleuropein may become a new adjuvant anti-ovarian cancer drug with potential applications.