Study On Anti-angiogenesis In Transplanted Human Breast Cancer Of Nude Mice By Metronomic Cyclophosphamide Combined With Trametes Robiniophila Murr

Objective：To assess the angiogenesis effect of metronomic cyclophosphamidecombined with Trametes Robiniophila Murr in nude mice with human breast cancer，toobserve the anti-tumor effect and the toxicity of the mice，to investigate the feasibilityof metronomic chemotherapy combined with Trametes robiniphila murr byanti-angiogenesis.Method：1. Established nude mice models of transplanted human breast cancer：Taking20female nude mice，human breast cancer cells line MCF-7were injected intothe right breast pad of each mice，0.2ml，6×10~7/ml.2. Divided into experimental groups and drug treatment: When the tumor volumewas about150-200mm3，the20tumor-bearing mice were randomly divided into4groups. Every group contained5nude mice. Group A (low-dose cyclophosphamide,LDM) CTX20mg/kg qd ip；Group B (maximum tolerated dose, MTD) CTX150mg/kg qod ip (3times)；Group C (low-dose cyclophosphamide combined with TrametesRobiniophila Murr) CTX20mg/kg qd ip，Trametes Robiniophila Murr200mg/d qd ip；Group D (control group) NS same volume qd ip.3. During the treatment，we observed the living conditions of nude mice every day，calculated the volumes of tumor and the weight of mice every2days and counted whiteblood cells in peripheral blood of nude mice every week.4. Tumor dissection and detection of pathology: All nude mice were killed bycervical dislocation after the21days treatments. We dissected the transplanted tumor，measured the weight of it and calculated the inhibition rate of tumor；Immunohistochemistry was used to detect the expression of microvessel density(MVD)、Vascular endothelial growth factor （VEGF）and thrombosponain-1(TSP-1) in different tumor tissues.Results：1. Inhibit tumor’s growth: When the7th days treatment，the growth of tumor ingroup C was significantly slowly compared with group A and group D (P＜0.05). After14days treatment，the growth of treatment groups were more slowly than group D(P＜0.05).Tumor volume of group C was significantly smaller than the group A (PAB=0.01)and the group C (PBC=0.024).After21days treatment，the growth of treatment groupswere significantly slowly compared with group D(P＜0.05).Tumor volume of group Cwas smaller than the group A(PAB=0.034)and the group C (PBC=0.023).Tumor weight ofcombined treatment group was the smallest，and the inhibition ratio of the tumor weightwas62.60%.2. Side effects: After7days treatment，the eating of nude mice in group B weredecling day by day，and the weight of mice was lost. After13days treatment，the weightof mice in group B was significantly slowly，（PAB=0.07，PBC=0.031，PBD=0.012）. After21days treatment，the mice of each group were at good conditions and the differencebetween each group were not obvious(P＞0.05).The statistical analysis of the leukocytecount showed there were not different among every experimental group.3. The results of immunohistochemistry: In the combination group and LDMgroup，xenografts expressed lower level VEGF and MVD and higher level TSP-1thanthose in the other groups(P＜0.05)，but there is no significant difference between thetwo groups(P＞0.05). Furthermore，there is no significant difference between MTD andNS in the expression of VEGF、MVD and TSP-1.Conclusion:Metronomic cyclophosphamide combined with Trametes robiniphilamurr has angiogenenic effect in immunodeficient mice bearing MCF-7human breastcarcinoma cell lines，and alleviate side effects of conventional chemotherapy.