Artemisinin Anti-angiogenic Effect On Canine Mammary Tumor Xenograft In Nude Mice

Breast cancer is a high incidence disease in humans,and the incidence of canine breast tumors is far higher than that of humans.The malignant rate is as high as 50%.At present,the treatment of canine mammary tumors is mostly combined with surgery and hormone chemotherapy.However,the results are often recurrent and have a large number of side effects.Artemisinin is an important extract that is widely used as an antimalarial drug.As the research progresses,its antitumor activity gradually becomes apparent.This experiment established a canine mammary tumor xenograft model in nude mice to investigate whether artemisinin has antitumor angiogenesis in vivo and possible mechanism.CHMm of the canine mammary gland tumor cells in logarithmic growth phase was taken.32 nude mice were injected with 0.2 ml of cell suspension of 2×10~7cells/ml respectively.The CHMm nude mice transplanted tumor model was successfully established 10 days after the injection.The nude mice models were randomly divided into four groups of 8 in each group.Each group was given daily gavage,high dose group:200mg/kg/0.1ml,middle dose group 100mg/kg/0.1ml,low dose group 50mg/kg/0.1ml,control group:0.1 ml vegetable oil was fed continuously for 21 days,nude mice body weight and transplanted tumor volume were recorded every 3 days,and nude mice were sacrificed on the 22nd day(24h after the last administration).The transplanted tumor tissue was dissected.Weigh tumor mass to calculate tumor inhibition rate;plot the tumor volume change curve during 21 days of continuous drug use;HE staining was used to observe pathological changes of the transplanted tumor;immunohistochemistry CD31 labeling neovascularization was used to measure microvessel density MVD;ELISA was used to detect serum vascular endothelial growth The content of factor VEGF and hypoxia-inducible factor HIF-1?were detected.The expression of Notch pathway-related factors in tumor tissue was detected by fluorescence quantitative assay.1.By comparing the changes in body weight of nude mice before and after treatment in each group,it can be seen that there was no significant difference between the groups before treatment and nude mice(p>0.05).After the end of treatment,except for the control group,the body weight of nude mice decreased significantly.There was no significant change in the body weight of nude mice in the artemisinin group(p>0.05).The three concentrations of artemisinin used in this study did not produce obvious toxic side effects to the body.2.After administration,the average mass of transplanted tumors in control group,50mg/kg,100mg/kg and 200mg/kg were:0.688±0.27g,0.521±0.23g,0.376±0.12g,0.306±0.55g,tumor inhibition The rates were:0,24.23%,45.34%,55.52%,and the difference was significant compared with the control group(p<0.05).The average volume of transplanted tumors in each drug group was smaller than that of the control group,and the 50 mg/kg drug group was significantly different.(p<0.05),the difference between the 100 mg/kg and 200 mg/kg groups was extremely significant(p<0.01).High-dose artemisinin group had better tumor inhibition than other groups.3.HE staining was performed on the transplanted tumor tissue.The control group showed less intercellular substance and more mitotic figures.In the artemisinin group,large area of tissue necrosis was seen,and the cells dissolved or even disappeared.It is uniformly red.4.The transplanted tumor tissues were immuno-histochemical CD31 markers,which were marked into brown and yellow neovascularization,and the blood vessel counts were counted.The microvascular density of the control group and the low,middle and high dose groups were 22.11±1.49,19.55±1.59,14.11±1.23 and13.56±1.29 respectively.The difference between the middle dose group and the high dose group was very significant compared with the control group(p<0.01).5.ELISA results showed that the serum VEGF decreased significantly in the high-dose group compared with the control group(p<0.01),while the other dose groups did not have significant(p>0.05);the serum HIF in the high-dose group compared with the control group The decrease in-1?was significant(p<0.05),and the other groups were not significant(p>0.05).6.The result of fluorescence quantitative section showed that artemisinin could down-regulate the expression of notch signaling related factors notch1,Dll4 and Jagged1,and 200mg/kg dose group had the most significant effect.CONCLUSION:Artemisinin has inhibitory effect on canine mammary tumor CHMm xenografts in nude mice.It may inhibit the development of tumors by reducing serum angiogenesis-related factors VEGF,HIF-1?and inhibiting the activity of notch signaling pathway related factors.