Mechanism Of Melittin Inhibits The Proliferation And Migration Of Osteosarcoma 143B Cells

Objective:The inhibitory effect of melittin on the proliferation and migration of human osteosarcoma 143 B cells was investigated by in vitro and in vivo experiments,and the mechanism of action was further explored.Providing a reliable experimental basis for the treatment of osteosarcoma with melittin.Methods:1.In vitro studies: 1)CCK-8 method to detect the effect of melittin on the proliferation of osteosarcoma 143 B cells;2)Wound healing experiment and Transwell chamber test to detect the effect of melittin on the migration ability of osteosarcoma 143 B cells;3)Flow cytometry was used to detect the effect of melittin on apoptosis of osteosarcoma 143 B cells;4)Realtime RT-PCR and Western blot were used to observe the effects of melittin on gene and protein levels of key factors LRP5,?-catenin,c-Myc and Survivin in Wnt/?-catenin signal transduction pathway in osteosarcoma 143 B cells.2.In vivo studies: Mice with established osteosarcoma tibial implants,observing the growth of melittin on transplanted tumor in nude mice.The morphology of osteosarcoma was observed by HE staining.The expressions of LRP5,?-catenin and Survivin in nude mice were detected by immunohistochemistry.Results:1.In vitro experiments: Melittin inhibits the proliferation of human osteosarcoma 143 B cells,and its inhibitory effect on cell proliferation in a concentration-dependent manner.Wound healing and Transwell chamber assay showed that the migration and invasion ability of cells gradually decreased with increasing concentration.Flow cytometry experiments showed that melittin significantly promoted apoptosis of osteosarcoma cells in a concentration-dependent manner.Realtime RT-PCR and Western Bolt assay showed that after treatment with melittin on 143 B cells,the expression of LRP5,?-catenin,c-Myc,Survivin m RNA and protein were gradually down-regulated.2.In vivo experiments: Melittin inhibits the growth of tibia xenografts in nude mice of different degrees.The middle dose group and the high dose group can effectively inhibit tumor growth.HE staining results showed that the tumor tissues treated with melittin showed different degrees of nuclear pyknosis.At the same time,the results of immunohistochemistry assay showed that the expression of LRP5,?-catenin and Survivin was underexpressed in the tumor tissue of each group of melittin was compared with the control group.Conclusions:Melittin inhibits the proliferation and migration of human osteosarcoma 143 B cells and induce apoptosis by regulating the expression of Wnt/?-catenin signaling pathway-related factors in vitro,and inhibiting the growth of xenografts in nude mice in vivo.