Inhibitory Effect Of The Combined Application Of Kangaifangzhuatang And 5-FU On The H22 Bearing Miceand Possible Mechanisms

Objectives: To investigate the effects of 5-Fluorouracil (5-FU), kangaifangzhuant and the combination of 5-FU and Kangaifangzhuantang respectively on the hepatoma bearing mice and the possible mechanisms by detecting the expression profiles of BAX,VEGF,EGFR under the different conditions mentioned above.Methods: The model of hepatocarcinoma solid tumor-bearing Kunming mice was established by:(1)H22 tumor cells were obtained from the abdominal dropsy of hepatocarcinoma 22 solid tumor-bearing Balb/c mice and then diluted to a final cell density of 10~6 cells per m L of cell suspension;and(2)0.2mL of diluted H22cell suspension was inoculated subcutaneously at the right armpit of each Kunming mouse.The model is divided randomly into four groups and they are normal saline control group, Kangaifangzhuantang group,5-FU group, and the combined application of Kangaifangzhuantang and 5-FU group.After fed with drugs for 14 days, the mice were killed and the tumor tissues were taken out for the continuing detections.And the detections include:(1)the weight of the tumor tissues;(2)tumor inhibition rate;(3) regular pathological observation by HE staining;and (4)the protein expression profiles of BAX,EGFR and VEG respectively by Immunohistochemistry among different groups.Conclusion: 1.There is no significant difference in tumor inhibition rate between Kangaifangzhuantang and 5-FU group(P﹥0.05), while the tumor inhibition rate is strikingly higher in the combined application of Kangaifangzhuantang and 5-FU group than Kangaifangzhuantang and 5-FU groups(P﹤0.05).2.the positive expression percentage of BAX protein is significantly lower in control group than Kangaifangzhuantang,5-FU group and the combination of Kangaifangzhuantang and 5-FU group(P﹤0.05).The positive expression percentage of BAX protein is the highest and is obviously higher than that of Kangaifangzhuantang and 5-FU group(P﹤0.05)and there was no significant difference in the positive expression percentage of BAX between Kangaifangzhuantang and 5-FU groups.(P﹥0.05).3.the positive expression percentage of VEGF protein is significantly higher in control group than Kangaifangzhuantang,5-FU group and the combination of Kangaifangzhuantang and 5-FU group(P﹤0.05). The positive expression percentage of VEGF protein is the lowest and is obviously higher than that of Kangaifangzhuantang and 5-FU group(P﹤0.05)and there was no significant difference in the positive expression percentage of VEGF between Kangaifangzhuantang and 5-FU groups. (P﹥0.05)4.the positive expression percentage of EFGR protein is significantly lower in control group than Kangaifangzhuantang, 5-FU group and the combination of Kangaifangzhuantang and 5-FU group(P﹤0.05). The positive expression percentage of EFGR protein is the highest and is obviously higher than that of Kangaifangzhuantang and 5-FU group(P﹤0.05)and there was no significant difference in the positive expression percentage of EFGR between Kangaifangzhuantang and 5-FU groups. (P﹥0.05)Results: Kangaifangzhuantang whether used solely or combined with 5-FU, has an inhibitory effect on tumor growth and the combination group is the most effective. The possible mechanisms may include by but not constraint to:(1)up regulation of the protein expression level of BAX and thus promoting cell apoptosis in the solid tumor;(2)down regulation of the protein expression level of EGFR and further inhibiting cell proliferation;(3)down regulation of the protein expression level of VEGF and so attenuating the angiogenesis in solid tumor. On the basis of all these experiments, it suggests that the Kangaifangzhuantang has the anti-tumor effect and does better when combined with the 5-FU. These observations provide a solid laboratory basis for the clinical application of the Kangaifangzhuantang, and it may action as a principle for the clinicians when it comes to the different application methods of Kangaifangzhuantang.