Study of Caesalpinia Sappan Aqueous Extract Inhibiting Effects in Human Ovarian Cancer Cell

Abstract: Objective:To observe caesalpinia sappan aqueous extract's growth inhibiting effects and investigated the mechanism of apoptosis-inducing on human ovarian cancer cell line SKOV3.Establishing the implantation nude mice models of ovarian cancer, so as to observe the caesalpinia sappan aqueous extract's elicit antitumor immunity in ovarian cancer model established in nude mice.Methds:Cell growth inhibiting effect was detected by cytometry; To examine cells morphological changes after the treatment by trypan blue staining under a light microscope; Electronic microscopy was used to observe ultrastructure of SKOV3 cell effected by caesalpinia sappan aqueous extract; With different concentration of caesalpinia sappan aqueous extract acting on cells, cells cycle and apoptosis rate were analyzed by flow cytometry (FCM). With different concentration of caesalpinia sappan aqueous extract acting on cells, immunocytochemistry was used to detect the expression of Caspase-3, Caspase-9 and survivin assay in ovarian cancer cell line SKOV3. SKOV3 cell line was inoculated subcutaneously and intraperitoneally into nude mice, and both models were randomly divided into three groups: control group, caesalpinia sappan aqueous extract group, DDP group. Tumor formation in mice was observed every 3 days, recorded the length of the tumor formation, measured the length and width so as to calculate the volume of tumor in the models of subcutaneously implantation. Recorded survival time of every intraperitoneal implantation mice, compared of the mean survival time of control groups with caesalpinia sappan aqueous extract groups and DDP groups.Result:Caesalpinia sappan aqueous extract can effectively inhibit growth of SKOV3; morphology typical of apoptosis (cell shrinkage, chromatin condensation and apoptotic body formation) were observed and in a concentration-dependent manner. Flow cytometry detection showed:the in vitro experiment revealed growth of SKOV3 was inhibited after treatment by caesalpinia sappan aqueous extract at different concentration. Apoptosis rate was increased with the increasing of the concentration of caesalpinia sappan aqueous extract.Immunocytochemistry showed that:positive rate of Caspase-3, Caspase-9 increased with the increasing of the concentration of caesalpinia sappan aqueous extract, while the gray scale lowered. Positive rate of surviving decreased with the increasing of the concentration of caesalpinia sappan aqueous extract, while the gray scale increased. At the concentration of 0、25μg/ml,50μg/ml, 100μg/ml,200μg/ml:the positive rate of caspase-3 were 5.29%,12.61%, 24.60%,100%,100%; the positive rate of caspase-9 were 12.32%,29.17%,57.42%,100%, 100%; the positive rate of survivin were 81.62%,60.53%,36.94%,55.69%,36.92%.Animal experiment results show that in Subcutaneous-implantation model, the mean volume of control groups was 0.545±0.134cm3, caesalpinia sappan aqueous extract groups and DDP groups were respectively 0.304±0.088cm3,0.260±0.100cm3. Comparisons of the mean volume of control groups with the volume of caesalpinia sappan aqueous extract groups and DDP groups have statistical significant difference. (P<0.01). As observation was terminated in the models of intraperitoneally implantation, the mean survival time of control groups was 62±8.97 days, caesalpinia sappan aqueous extract groups and DDP groups were respectively 81±7.12 days,90±0 days. Comparisons of the mean survival time of control groups with the survival time of caesalpinia sappan aqueous extract groups and DDP groups have statistical significant difference. (P<0.01).Conclusion:Caesalpinia sappan aqueous extract can obviously inhibit growth of human ovarian cancer cell line SKOV3, and induce apoptosis of it by increasing expression of Caspase-3, Caspase-9 and decreasing expression of survivin. The growth of the tumor in nude mice were markedly inhibited and the survival time of nude mice was extended by caesalpinia sappan aqueous extract in a certain dose…
Key words: human ovarian cancer cells; caesalpinia sappan aqueous extract; apoptosis mechanism; nude mice; animal experiment

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