The Effect Of Survivin On Prognosis Of Breast Cancer And Observation Of Apoptosis Of SK-BR-3 Cells Induced By Endostatin

Part 1:The relationship between expression of peripheral-blood Survivin and prognosis of advanced breast cancerObjective:1 To detect the Survivin expression in peripheral-blood and cancerous tissues of advanced breast cancer patients.2 To study the relationship between expression of Survivin in peripheral-blood and expression of Survivin, Her-2, hormone receptor in cancerous tissues of advanced breast cancer patients.3 To study the relationship between expression of peripheral-blood Survivin and response to chemotherapy or prognosis in advanced breast cancer patients.Methods:1 Expression of peripheral-blood Survivin was semi-quantitied determined by the methods of RT-PCR.2 Expression of peripheral-blood Survivin was quantitied determined by the methods of real-time quantitative PCR.3 Expressions of Survivin and Her-2 in cancerous tissues were determined by the method of immunohistochemistry.4 Analyze the clinical characteristics of 15 advanced breast cancer patients: sexuality, age, pathology, clinical stage, respond rate.5 Data processing.Results:1 The positive expression rate of Survivin is 10/15 (66.7%).The Survivin expression detected by immunohistochemical examination coincided with the method of RT-PCR.2 The clinical disease control rate of patients treated with TP regimen was 11/15 (73.3%), 4 patients who achieved PD were SurvivinmRNA positive expression and had higher levels of SurvivinmRNA expression after chemotherapy.3 Her-2 protein positive expression rate was 6/15(40.0%), ER/PR positive expression rate was 8/15(53.3%).SurvivinmRNA in peripheral-blood may have correlation with Her-2 and ER/PR.Conclusions:SurvivinmRNA. in peripheral-blood may correlate with Her-2 and ER/PR. Survivin may become one of the important tumor markers to predict therapeutic effect of TP regimen and prognosis of advanced breast cancer patients.Key words: Breast cancer Survivin PrognosisPart 2:The effects of both endostar and docetaxel on the growth of SK-BR-3 cell proliferation and their mechanismsBackground:EndostarTM (rh-endostatin)is a new recombinant human endostatin developed by Jiangsu Simcere Pharmaceutical Co. Ltd Nanjing. P,R. China. Pre-clinical study indicated that endostar could inhibit tumor endothelial cell proliferation, angiogenesis and tumor growth. Recently, it was reported that endostar could also inhibit colon cancer cell proliferation directly. Our prophase research indicated that endostar might induce lung cancer cell apoptosis in spite of the mechanism is not clear.Objective:1. To observe the effects of endostar and docetaxel on SK-BR-3 cells proliferation and investigate the apoptotic effects on SK-BR-3 cell and apoptosis associated proteins.2. To assess the combination effects of endostar and docetaxel on SK-BR-3 cells proliferation. If synergistic effect was observed, we further investigate the optimist time or dose of the combination regimen and the possible mechanism.Methods:1. The effects of endostar and docetaxel on SK-BR-3 cells proliferation were studied by means of MTT.2. The cell apoptosis and cell cycle were determined by transmission electron microscope and cell flow cytometry.3. The expression of apoptosis associated gene were analyzed by enzyme linked immunosorbent assay(ELISA).Results:1. The inhibitory effect of endostar on growth of SK-BR-3 cells was time but not dose dependent. The growth inhibitory rate of SK-BR-3 cells was 30.1% after treatment with endostar at concentration of 1μg/ml for 72h. The apoptotic rate was 11.6% and cell cycle was not influenced. The expression of sFas and sFasL were not detected in SK-BR-3 cells. The expression of Bcl-2 decreased (16.64% vs 10.32 %);while the expression of Bax did hOt,change significantly(2.65 % vs 2.53%).2. The inhibitory effect of docetaxel on growth of SK-BR-3 cells was time and dose dependent. The growth inhibitory rate of SK-BR-3 cells was39.4 % after treatment with docetaxel at concentration of 2μg/ml for 24h. The apoptotic rate was 12.40% and the percentage of cells accumulated in G2/M enhanced significantly .The expression of sFas and sFasL were not detected in SK-BR-3 cell too. The expression of Bcl-2 decreased (16.64% vs 12.71%);while the expression of Bax enhanced but not significantly(2.65% vs 3.24%).3. Simultaneous endostar plus docetaxel produced a synergistic effect. However, the growth inhibitory rate of sequential administration did not enhance significantly compared with docetaxel. Simultaneous administration showed best apoptotic effect which may attribute to the decreaced expression of Bcl-2.Conclusions:Both endostar and docetaxel could inhibit proliferation of SK-BR-3 cells, induce SK-BR-3 cells apoptosis. Simultaneous administration exhibited best inhibitory effect. The synergistic effect was associated with decrease of Bcl-2.