Research On Chemotherapy By 5-Fu And CIS Platin On Rabbits Model Induced By VX-2 Cells

Objective:1.The objective of the present investigation was to explore whether the combination of routine preoperative intestinal lavage with intestinal cavity chemotherapy could achieve better curative effect on colorectal cancer. In order to fulfill this objective, a model on colostomy for colorectal cancer on rabbit(hereinafter referred to as “model”) was established, and an experiment of chemotherapy bath of tumor was performed to simulate the intestinal cavity perfusion chemotherapy. 2.Detect zhe VX-2 cell viability by use MTT, observe zhe curative effect of Polyethylene glycol combined with 5-fu or DDP to colorectal cancer on rabbit.Methods:Part one: Preparation before the establishment of model: Healthy rabbits, regardless of their gender, weighing 1.8-2.5kg, were selected. Colostomy was conducted on the rabbits. after operation, the model on rabbit was prepared by using injection method of tumor cell suspension. The growth of tumor was observed, and size of fistula tumor was measured every day after model construction. Twenty rabbit models with similar sizes of tumors were selected and randomly divided into 4 groups.Four groups of rabbits were divided into 3 treatment groups and 1 control group: polyethylene glycol bath group(group A, n=5), polyethylene glycol combined with 5-Fu(0.75g/L) chemotherapy bath group(group B, n=5), polyethylene glycol plus cisplatin(400 mg/L) chemotherapy bath group(group C, n=5), polyethylene glycol combined with 5-Fu(0.75 g/L) and cisplatin(400 mg/L) chemotherapy bath group(group D, n=5).Two times of chemotherapy bath was conducted in the treatment group with an interval of 3 days. Routine blood test, electrolytes, and liver and kidney function tests were conducted one day before chemo-bath and after it. Changes that were before and after chemotherapy bath were compared. Tumor tissues were collected for pathological examination. The expression levels of vascular endothelial growth factors(VEGF) and proliferating cell nuclear antigens(PCNA) in various groups were examined and compared by immunohistochemical method.Part two:Vaccinate the combination of the polyethylene glycol, different concentration of 5-Fu, cisplatin and VX-2 cell suspension 200μl into the 96-hole plate, set a blank control group(1640 nutrient fluid) and eight drug treatment groups, set four vice-blank in each group. After 24-hour’s cell culture, add the drug 20μl with different thickness into the treatment groups, then put them into the 370 C incubator. Add MMT 20μl after 24-hour’s cell culture, then observe the survival rate of the cells after 24-hour’s function from the different concentrations of the drug.Results:1. In three weeks,There were tumor foci in fistula of about 70% rabbits implanted with tumor after operation, indicating that the tumor cell suspension injection method could be successfully used to establish a rather stable model.2. There was no statistical significance(P > 0.5) in blood routine indexes, such as hemoglobin, white blood cells and red blood cells, liver and kidney function indexes, such as total proteins, glutamic-oxalacetic transaminase, glutamic-pyruvic transaminase and serum creatinine, and other electrolytes indexes, such as sodium, potassium and calcium in various treatment groups before and after soaking tumor in chemotherapy. The experimental animals in each group exhibited decrease of appetite and weight within 10 days of the said operation.3.Life cycles of animals in the treatment groups are longer than those in the control group. Animals in group D have the longest life cycle. The significance between group A and group B/C/D was P<0.05, respectively. The significance between group D and group B/C was P < 0.05, respectively.4. Gray-scale VEGF: the gray-scale values of VEGF in group B, group C, group D were higher than that of group A. The significance between group A and group B、C、D was P <0.05, respectively. The significance between group D and group B<C was P < 0.05, respectively.5. Gray-scale value of PCNA: the gray-scale values of PCNA in group B, group C, group D were higher than that of group A. The significance between group A and group B/C/D was P<0.05, respectively. The significance between group D and group B/C was P < 0.05, respectively.6.Polyethylene glycol combined with different concentration of 5-Fu and cisplatin had a certain inhibition on VX-2 cells after 24 hours.Conclusion:1. The success rate of establishment of the model by using the VX-2 cells is high, and the model is stable enough to satisfy the requirement of the experiment.2. The dissolution of 5-Fu, cisplatin, and 5-Fu combined with cisplatin in polyethylene glycol solution is proved to be a safe and effective method for chemotherapy bath of tumor, which has direct killing effect on tumor cells and prolongs the survival time of the experimental animals.The curative effect of cisplatin combined with 5-Fu on chemotherapy bath of tumor is better than that of the single drug group.3.In vitro experiment, the lethal effect of 5-Fu and cisplatin on VX-2 cells is in proportion to the thickness, synergistic action can be seen when these two ingredients are combined together.