The Treatment Of Photodynamic Therapy Combination With Fluorouracil Implants Experiment On H22 Hepatoma-bearing Mice

Objective: Hepatocellular is a common malignancy worldwide and one of the greatest threats to human existence. The incidence of hepatocellular in the world is rising. According to the World Cancer Report 2014 published by World Health Organization, in 2012, approximately half of the new cases of hepatocellular occur in China which is the largest number of new cases and deaths in the world. Although the existing therapies are able to benefit patients with hepatocellular carcinoma to some extent, traditional treatments still have limitations. Photodynamic Therapy(PDT) is a technique for diagnosing and treating disease appeared in the late 1980 s. PDT has been developing rapidly in the treatment of neoplastic and non-neoplastic diseases because of the broad indications, trauma, low toxicity, targeting, repeatability. It can provide palliative care, eliminate occult lesions, be coordinated to improve the efficacy of surgery. It also can protect the appearance and function of vital organs, such as rapid recovery. Although technology is not yet mature, PDT has been used in the treatment of hepatocellular and the future is prosperous. Fluorouracil Implants, under the trade name of Sinofuan, is a new type of sustained-release implant, which makes of fluorouracil as raw materials, and biocompatibility of medical polymer materials as skeleton. The local drug concentration released by Sinofuan is 10 times more than intravenous drug use, and the effective drug concentrations can maintain above 240 h. Sinofuan can elevate the local drug concentration and extend the time of effective drug concentration in it’s particular mode to achieve a ideal administration mode with strong antineoplastic efficacy and low side effect. The subject of this study is H22 hepatoma-bearing Kunming mice model. In the study, the researcher conducts the treatment of Photodynamic therapy combination with fluorouracil implants experimental on H22 hepatoma-bearing mice, in order to improve the local control rate and prolong survival of hepatocellular. The study will provide a theoretical basis for the treatment of photodynamic therapy combined with local chemotherapy using in the clinic.Method:1 Create transplantation tumor model of H22 hepatoma mice and divide into four groups randomly: control group(C)、fluorouracil implants group(SF)、photodynamic therapy group(PDT)、combination therapy group(PDT+SF). Group C are not given any treatment; Group SF are implanted 100mg/Kg fluorouracil implants intratumor; Group PDT are given PDT; PDT+SF group are received implantation of 100mg/kg fluorouracil implants intratumor and PDT.2 Every other day to measure transplantation tumor of mice, to calculate the tumor volume and tumor growth inhibition rate and growth curve; To observe the growth and body weight change of mice before and after treatment.3 Part of the mice are sacrificed after 14 days, stripping out the tumor and weight it, to calculate tumor inhibition rate. Remove the spleen of mice and weight it, to calculate spleen index and to analyze the effect of each treatment on the immune system of mice.4 The remaining mice are observed survival time and quality of life, to draw survival curves and to calculate the life span.5 To observe the results of HE staining pathological section from transplanted tumor by microscope.6 Flow cytometer detect the apoptosis and cell cycle distribution of tumor.7 Statistical analysis: SPSS19.0 statistical software for data analysis. Measurement data is expressed with x ±s. Each group of indicators are compared using ANOVA test, each group survival of tumor-bearing mice were analyzed using Kaplan-Meier method. P<0.05 are statistically significant for each test.Results:1 The formation rate of H22 hepatoma transplanted tumor is 100%,simple and reproducible to operate.2 Changes in body weight of mice: The body weight of mice is no significant difference in each group by analysis of variance before treatment(P>0.05). The first 14 days after treatment,the weight in group C、SF、 PDT、PDT+SF are:(35.58±2.04),(31.79±1.76),(32.46±1.88),(33.20±1.85). Group C and group SF, group C with group PDT are statistically significant by analysis of variance(P<0.05). The difference between the other groups is not statistically significant(P>0.05).3 Changes in transplanted tumor volume of mice and tumor growth inhibition rate: the average initial tumor volume is(792±41) mm3 before treatment, there is no significant difference among groups by analysis of variance(P>0.05). According to statistic analysis, there is significant difference among the groups(P<0.05). The tumor growth inhibition rates of group C、SF、PDT、PDT + SF are: 0,51.62%, 43.35%, 68.68%.4 Each group transplanted tumor weight and inhibition rate: The tumor weights of group C、SF、PDT、PDT + SF are:(5.42±0.27) g,(2.69±0.34) g,(3.05±0.37)g,(1.72±0.27).The tumor weight of group PDT+SF is significantly lower than the group SF and group PDT, the difference is statistically significant(P<0.05), there is no significant difference between group SF and group PDT(P=0.094).The tumor inhibition rate of group C、SF、PDT、PDT+SF are: 0,50.37%, 43.73%, 68.27%.5 Survival of tumor-bearing mice: the treatment groups can prolong the survival time of tumor-bearing mice, improve the quality of life of mice, which group PDT+SF has the longest survival time and the highest life span.6 The pathologic observation for transplanted tumor: Compared with the group C, the treatment groups are appeared to reduce the number of tumor cells and pathological mitotic,necrosis and vacuolar degeneration can be seen, PDT + SF group shows the largest necrosis area.7 Apoptosis and cell cycle distribution: Apoptosis rate has increased in group SF、group PDT and group PDT+SF Compared with group C, the most increasing can be seen in group PDT + SF with the highest rate of apoptosis, the differences among the groups are statistically significant(P=0.000). Cell cycle distribution showed that the cell proportion of G0/G1 phase in group SF、group PDT and group PDT+SF are risen compared with group C, but for S phase is decreased. Group PDT+SF shows the most obviously decline(P=0.000).8 Spleen index detection for each group of mice: the spleen index of group SF is declined compared with group C. the difference is statistically significant(P<0.05).There is no significant difference between group PDT、group PDT+SF and group C(P>0.05).Conclusion:Photodynamic therapy combination with fluorouracil implants can inhibit tumor growth effectively, prolong survival time of tumor-bearing mice, improve the life quality of the mice, promote the apoptosis, and doesn’t show the risk of reduction of immunity.