Mechanism Of Yiliu Decoction Regulating Malignant Proliferation Of Osteosarcoma Through IFITM1/Wnt/β-catenin Pathway

Objective:1.Through the experiment in vitro,the effect of Yiliu decoction containing serum on the growth of osteosarcoma U2-OS cells was investigated.2.To investigate the effect of Yiliu Decoction on body weight,tumor volume,tumor weight and tumor inhibition rate in nude mice,and to detect the expression of IFITM1Wntβ-catenin protein m RNA in tumor,so as to explore the possible relationship between IFITM1Wnt/β-catenins in the treatment of osteosarcoma.Methods:1.In vitro experiment:Use Yiliu Decoction to intervene U2-OS cells and observe the changes of proliferation,invasion and migration,and apoptosis of U2-OS cells after intervention.2.In vivo experiment:30 SPF nude mice were selected,checked the random number table,and randomly assigned to 3 groups,with 10 mice in each group.They were divided into Yiliu decoction group,cisplatin group and normal saline group.Each nude mouse was injected into the tibia of the left hind limb with a concentration of 1×0.1ml U2-OS cell suspension of10~7cells/ml was used to construct nude mice transplantation model of osteosarcoma.After successful modeling,the tumor body grew to 0.5cm x0.5cm and began to be administered.The volume of subcutaneous tumor and body weight of nude mice in each group were recorded every 5 days.After 25days,the nude mice died suddenly,weighed the tumor weight and calculated the tumor inhibition rate of each group.The samples were used for Western Blot and q RT-PCR tests to detect the expression of related m RNA and protein.Results:1.Effect of Yiliu decoction on U2-OS cells:Yiliu decoction containing serum can inhibit the proliferation,invasion and migration of osteosarcoma U2-OS cells and promote apoptosis.2.1.Changes in tumor volume,weight and tumor inhibition rate of nude mice in each group after administration of nude mice osteosarcoma transplantation model:compared with normal saline group,the tumor volume of tumor inhibition decoction group and cisplatin group decreased significantly.The tumor inhibition rates of Yiliu decoction group and cisplatin group were 32%and 55%respectively,and the tumor weight of Yiliu decoction group and cisplatin group was lower than that of normal saline group.The body weight of nude mice in the normal saline group was smaller than that in the tumor suppression decoction group and cisplatin group.2.2.The expression of IFITM1/Wnt/β-catenin m RMA in osteosarcoma transplantation model of nude mice:Compared with NS group and cisplatin group,the expression of IFITM1 m RNA in Yiliu Decoction group had no significant difference between NS group and cisplatin group;Compared with NS Group,the expression of Wnt m RMA in Yiliu Decoction Group and Cisplatin Group increased;The expression ofβ-catenin m RMA in Yiliu Decoction Group and Cisplatin Group was higher than that in NS Group.2.3.Expression of IFITM1/Wnt/β-catenin protein in osteosarcoma transplantation model of nude mice:Compared with NS group and cisplatin group,the expression of IFITM1 protein in Yiliu Decoction group had no significant difference between NS group and cisplatin group(P<0.05);Compared with NS group,the expression of Wnt protein in Yiliu Decoction group and cisplatin group increased.Compared with NS group,the expression ofβ-catenin protein in Yiliu Decoction group and cisplatin group increased.Conclusion(s):1.In vitro and in vivo,it was found that Yiliu Decoction could inhibit the growth of osteosarcoma U2-OS and improve the quality of life of nude mice with osteosarcoma.2.Yiliu Decoction may inhibit the growth of osteosarcoma by inhibiting the expression of IFITM1 and causing the high level of Wnt/β-catenin...