Influence Of Kuilinghuajijian On Anti-tumor Actions And On C-myc And Nm23-H₁Expression In Mice Bearing Hepatocarcinoma H₂₂ Cells-derived Tumor

Objective: Cancer has become one of the main causes of death in the world, andhepatoma is one of the most common malignant tumor. Main means of treatment， suchas：surgery、radiotherapy and chemotherapy have some certain limitations, especiallythe toxic effects of chemotherapy drugs will reduce the quality of life of the cancerpatients. So effective traditional Chinese medicine with low toxicity has gradually beenused in the treatment of various tumors. In this study, by fluorouracil as a positivecontrol and sodium chloride as a negative control, it will observe the antineoplasticaction of Kuilinghuajijian in the mice bearing H₂₂tumor cell and explore its possiblemechanism of action.Methods: In order to establish hepatocellular carcinoma solid tumor model in vivoexperiments, following steps were taken. First, resuscitated frozen hepatocellularcarcinoma successfully, and made continous cell culture in Kunming mice. Then,extracted ascites from these mice in a sterile environment,and diluted it to concentration5x106a mL﹣1H₂₂. Last, vaccinated suspension0.1mL in the right armpits of64micerespectively,which were6to8weeks old, weighing18²2g. These64mice wererandomly divided into4group: the control group （0.85%saline0.4mL）,5-Fu group（Fluorouracil20mg/kg）, traditional Chinese medicine group （Kuilinghuajijian0.4mL）and the combination group （Kuilinghuajijian0.2mL plus fluorouracil10mg/kg）,16mouse in each group,8male and female each.4groups of mice receive daily gavage orintraperitoneal injection medication respectively. During continuous administration oftreatment, the changes in the state of the survival of mice in each group were observed and recorded, After14days, the mice were sacrificed by cervical dislocation method,and the transplanted tumors were stripped, then photographed and weighed. The effectof the inhibition of liver cancer cells was observed by comparing the transplantedtumor’s volume, weight, inhibitory rate in each group, and the expression of C-myc andnm23-H₁was detected by using immunohistochemistry technique.Results: With the time of treatment, the mice in traditional Chinese medicinegroup better quality of life than the combined group and5-Fu group, through observingthe index of their mental status, respiratory rate, hair color, body shape, food intake andactive sensitivity in mice medication during treatment. By comparing the size of micetransplanted tumor in4groups, it was clearly observed that the size of5-Fu group,traditional Chinese medicine group and the combination group was less than the controlgroup, and the5-Fu group is the most obvious one in inhibiting tumor growth, and thecombination group is more obvious than the Chinese Medicines in the antitumor effect.Judging from the weight of the transplanted tumors, the inhibition ratio was50.23%inhepatoma cells by using5-Fu, it was36.41%by using Kuilinghuajijian combined with5-Fu and it was29.03%by using Kuilinghuajijian, which were obviously lower thancontrol group （﹡﹡P<0.05,﹡P<0.05）.The expression of C-myc and nm23-H₁in fourgroups were positive from the results of immunohistochemical. The statistic of ANOVAshows that Kuilinghuajijian can weaken the expression of C-myc and strengthen theexpression of nm23-H₁.4treatment groups are all lower than in controls（△P<0.05）. Butno statistical significant among the three treatment groups （P>0.05）.Conclusions: Kuilinghuajijian and fluorouracil both play a significant role ininhibition of tumor growth, its mechanism of action may be related to the regulation ofthe expression of C-myc and nm23-H₁.Because of the much lower toxicity ofKuilinghuajijian, it can improve the quality of life of cancer patients, and prolong thesurvival of patients with cancer. Therefore, from the long-term efficacy, low toxicitydrug Kuilinghuajijian can be used as a new drug for the treatment of liver cancer.However, in this experiment, it just only preliminary clarify the possible mechanism ofanti-tumor, its clear mechanism still needs further discussion and research.