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The Effect of Rh-endostatin Combined with 5-fluorouracil of the Proliferation and Apoptosis in Choriocarcinoma JEG-3 Cells

Author: ChenWuJuan
Tutor: LiuHuiNing
School: Central South University
Course: Obstetrics and Gynaecology
Keywords: JEG-3 cells YH-16 Proliferation and apoptosis VEGF-B
CLC: R96
Type: Master's thesis
Year: 2011
Downloads: 40
Quote: 0
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Abstract


Objective: To observe recombinant human endostatin (Endostar, YH-16, Endostar), 5 - fluorouracil (5-fluorouraeil ,5-FU), and the combination inhibition of choriocarcinoma JEG-3 cell growth and induced apoptosis, and to explore the combination between the way the molecular mechanisms of synergy. Methods: (1) MTT assay was used to observe the different concentrations of YH-16 and 5-FU, and both combination therapy of highly invasive human choriocarcinoma cell growth inhibition rate in JEG-3 cells and at different times; (2) flow -cell assay YH-16 ,5-Fu monotherapy and combination JEG-3 cell apoptosis and cell cycle; (3) RT-PCR assay YH-16 ,5-Fu monotherapy and combination therapy JEG-3 cells, vascular endothelial growth factor VEGF-B expression. Results: (1) MTT method was used to observe the growth inhibition rate of the drug group JEG-3 cells and at different times. The same drug concentration, different drug group cell growth inhibition rates with the prolonged duration of action (24, 48, and 72 hours) and increases were statistically significant differences between groups, P lt; 0.05. YH-16 group of different concentrations: the same action time the 200ug/m1 group of cell growth inhibition rate in 50ug/ml, 100ug/ml group was statistically significant, P lt; 0.05; 200ug/ml with 400ug/ml group, was not statistically significant, P gt; 0.05; different concentrations of 5-FU (0.1mg/ml, 0.25mg/ml, 0.5mg/ml, 1mg/ml) group of cell growth inhibition rate and Drug concentration has a linear correlation, the correlation coefficient r gt; 0.9, P lt; 0.05 5-FU for 24, 48 and 72 hours, IC 50 were 1.56 mg / ml, 0.52mg / ml and 0.31 mg/ml.200ug / ml YH-16 Joint different concentrations of 5-FU (0.1mg/ml, 0.25mg/ml, 0.5mg/ml, lmg / ml) group role different times after cell growth inhibition rate is much higher than with the corresponding concentration of YH-16 and 5-FU group, P lt; 0.05, statistically significant. With increasing the concentration of 5-FU, the growth inhibition rate increases. 5-FU in the 24-hour, 48-hour and 72-hour IC50 were: 0.94mg/ml, 0.31mg/ml and 0.10mg/ml. The combination index CI values ??calculated for 24 hours, 48 ??hours and 72 hours, respectively 0.57,0.49 and 0.50, that the combination has a synergistic effect. (2) flow cytometry method associated with the YH-16 ,5-FU and both JEG-3 cell apoptosis and cell cycle. After each drug for 48 hours, 200ug/ml YH-16 group, 0.5mg/ml 5-FU group and the combination of the two groups of cells apoptosis rate was significantly greater than the control group, P lt; 0.05. The apoptosis rate of the combined treatment group was significantly higher than the 5-FU alone and YH-16 group, P lt; 0.05. Different the drug group Go/G1 phase cells percentage compared with the control group, the G2 / M phase cell percentage compared with the control group decreased, P lt; 0.05. The percentage of cells in G0/G1 phase of the combination group than YH-16 ,5-FU alone treatment group, P lt; 0.05. (3) RT-PCR assay YH.16 ,5-FU and their combination with VEGF-B expression in JEG-3 cells. 48 hours after drug intervention 200ug/ml YH-16 group, 0.5mg/ml 5-FU group, and both combined group compared with the control group, the expression levels of VEGF-B cells decreased, P lt; 0.05. The expression levels of VEGF-B of the cells of the combined treatment group was significantly lower than the individual drug group, P lt; 0.05. Conclusion: 1.YH-16 can directly inhibit the growth of JEG-3 cells, JEG-3 cells can be arrested in G0/G1 phase, to inhibit the proliferation of JEG-3 cells, induction of apoptosis, the mechanism may be associated with downregulation JEG 3 cells VEGF-B mRNA expression. 2.YH-16 joint 5-Fu treatment has a synergistic effect to inhibit the growth of JEG-3 cells, induce apoptosis, can improve the efficacy of 5-Fu chemotherapy.

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