| The incidence of ovarian cancer ranked third of gynecological malignancies, but its mortality is first. This is because ovarian is deep in pelvic cavity, most of the patients with onset of early symptoms, so it is difficult to detect and give the effective treatment. About60%-70%of patients with ovarian cancer are diagnosed at clinically advanced stages. The treatment of ovarian cancer is cells cytoreductive surgery combined with chemotherapy in the combined therapy. In recent years,although the application of paclitaxel and platinum-based drugs, making the five-year survival rate of ovarian cancer has increased, but ovarian cancer cells resistant to platinum drugs, highly recurrent advanced ovarian cancer, makes the five-year survival rate still hovering around20%to30%. Ovarian carcinomas have become one of the main threat to women’s health, so to find a way to increase the ovarian carcinomas cells to cisplatin sensitivity method to become one of the hot research now. Studies have shown that tumor development is not only related cells excessive proliferation, but also related with apoptosis is blocked. The imbalance of cell prolifertion and apoptosis, leading to the development of tumor. Therefore, the induction of more tumor cell apoptosis may delay the progress of the tumor.Cisplatin-induced apoptosis of tumor cells is achieved by an increase in intracellular of caspase-3expression. Smac gene is one of the promote apoptosis gene,the Smac ptotein which synthesis under the guidance has a role in promoting apoptosis,its overexpression in tumor cells,by increase of caspase-3expression, the induction of more malignant tumor cell apoptosis may delay the progression of malignant tumor development. The protein of SmacN7is one of the proposed synthetic Smac protein analogues, which keeps the natural biological activity of Smac protein. In this study, using cell-mediated immunity (SP method), methyl thiazoly tetrazolium (MTT) and colony formation assay, to investigate whether the SmacN7protein can increase the sensitivity of human ovarian caicinomas cells to cisplatin and the possible mechanisms, in ordor to provides a theoretical basis to advanced ovarian caicinomas. ObjectiveTo explore whether the mitochondrial pro-apoptotic polypeptides N7(SmacN7) can increase the sensitivity of human ovarian caicinomas cells to cisplatin and the relationship between the expression of Caspase-3protein. Methods1. Study Object:Logarithmic phase of growth of human ovarian caicinomas SKVO-3cells in vitro.2. Methods:Different concentrations of SmacN7protein combined with cisplatin and cisplatin alone which effect the culture of human ovarian caicinomas SKOV-3cells in vitro,Immunocytochemistry SP means to discuss the expression of Caspase-3protein in SKOV-3cytoplasmic after48h in different groups; after24h,48h,72h,using methyl thiazolyl tetrazolium (MTT) to detect the sffscts of cell proliferation;Colony formation of the experimental to study the rate of cell clones.3. Statistical method:Statistical analysis was done by SPSS17.0. One way ANOVA to analyst the datas.The statistical analysis of count data are ordinalvariables using the Chi-square test, and then spearman analysis the correlation; measurement data are expressed as mean±standard deviation. Different groups were compared using analysis of variance. Significant level being0.05. Results1. Compared to using cisplatin alone, SmacN7protein combination cisplatin application, the expression of Caspase-3protein can signicantlly increased (x2=40.663rs=0.45P=0.000)2. Using cisplatin alone,24h,48h,72h, cell proliferation inhibition rates were (6.15±1.12)%,(13.52±1.15)%and (29.31±1.07)%. SmacN7protein combination cisplatin application,50μg/L,100μg/L,and200μg/L, with the extend of the concentration of the increase of SmacN7protein or time longer,can significantly increased cell proliferation inhibition rate, after the drugs action24h,48h,72h later, the inhibition rates of proliferation were (9.26±1.73)%,(29.24±2.08)%and (48.97±2.09)%,(24.82±2.11)%,(43.12±1.72)%and (66.12±2.25)%,(43.95±1.23)%,(72.18±1.74)%and (81.97±2.17)%. SmacN7protein combination cisplatin application, SmacN7inhibited the proliferation of SKOV-3cells dependent on both time and concentration increase.Increase the concentration of SmacN7(50,100,200μg/L) and the role of the extension of time (24h,48h,72h), cell proliferation inhibition rate increased significantly. Same concentration at different time points cell proliferation inhibition rate difference was statistically significant (P<0.05); the same time at different concentrations cell proliferation inhibition rate difference was statistically significant (P<0.05).3. SmacN7protein combination cisplatin application;50μg/L,100μg/L and200μg/L, with the extend of the concentration of the increase of SmacN7protein, the cloned rate of SKOV-3cells were (21.17±0.55)%,(18.23±0.54)%,(15.27±0.56)%. Using cisplatin alone, the cloned rate of SKOV-3cells were (25.13±0.52)%. With the concentration of SmacN7(50,100,200μg/L), cell colony formation rate is gradually reduced. Difference was statistically signicant by statistical analysis (P<0.05). Conclusions1. The expression of Caspase-3protein increased in SKOV-3cytoplasmic, is the cause of SmacN7increased the sensitivity of human ovarian carcinomas SKOV-3cells to cisplatin, which is clutured in vitro.2. SmacN7protein combination cisplatin application, can enables the proliferation inhibition rate increased of human ovarian carcinomas SKOV-3cells which are cultured in vitro,the clone of cells decreased, so that can increased the apoptosis of tumor cells. |
PDF Full Text Request