Jatrorrhizine Inhibit Progression of Osteosarcoma by Downregulation of MAOB
Objective To investigate the effect of jatrorrhizine on the development of osteosar-coma and its mechanism.Methods The targets of jatrorrhizine on osteosarcoma were screened through network pharmacology analysis,and GSEA enrichment was used to analyze the relationship between the expression level of monoamine oxidase B(MAOB)and the clinical phenotypes of os-teosarcoma.The cell activities of osteosarcoma cells MG63 and U2OS were detected under different doses of jatrorrhizine(0,2,4,8,16 μmol/L),and the optimal jatrorrhizine dose for follow-up functional test was selected.CCK-8 method and Transwell assay were used to detect the cell prolifer-ation,migration and invasion of MG63 and U2OS,and Western blotting was applied to detect the protein expression levels of cell proliferation-associated antigen Ki67,matrix metalloproteinase 2(MMP2)and MMP9.The subcutaneous transplant models of osteosarcoma in nude mice were estab-lished,and the weight and volume of tumor tissues of each group were measured,and the expres-sion level of Ki67 was determined by immunohistochemistry.Western blotting was adopted to detect the effect of jatrorrhizine on the expression level of MABO protein in osteosarcoma cells.shRNA was used to interfere with the expression of MAOB,and its effect on the proliferation of osteosarcoma cells was analyzed.Results Ten targets of jatrorrhizine on osteosarcoma,including MAOB,was predicted by network pharmacology analysis.GSEA enrichment analysis showed that MAOB was re-lated to the prognosis(P＜0.05)and survival(log-rank P＜0.05)of osteosarcoma.The viability of MG63 and U2OS cells decreased with the increase of the drug dose.The dose of 8 μmol/L was chosen for the follow-up functional test as the viability of osteosarcoma cells decreased by about 50％when the dose of jatrorrhizine reached 8 μmol/L.In vitro experiment showed that compared with those in the Control group,the cell proliferation rate of jatrorrhizine group was significantly re-duced(P＜0.05),and the number of invasion cells and migrating cells were significantly de-creased(P＜0.05),and the protein levels of Ki67,MMP2 and MMP9 were significantly downreg-ulated(P＜0.05).In vivo experiment showed that the growth rate of subcutaneous tumors in nude mice of jatrorrhizine group was significantly reduced(P＜0.05),and the weight and volume of subcutaneous tumors were lowered(P＜0.05),and the expression level of Ki67 in tumor tissues was significantly declined(P＜0.05).Jatrorrhizine inhibited the expression of MAOB in osteosarco-ma cells in a time and concentration-dependent manner in vitro(P＜0.05).Knockdown of MAOB could significantly inhibit the proliferation of osteosarcoma cells when compared with shNC group(P＜0.05).Overexpression of MAOB could reverse the inhibitory effect of jatrorrhizine on the prolif-eration,invasion and migration of osteosarcoma cells(P＜0.05).Conclusion The expression lev-el of MAOB is related to the prognosis and survival of osteosarcoma.MAOB promotes cell prolifera-tion,invasion and tumor metastasis.Jatrorrhizine inhibits the proliferation,invasion and migration of osteosarcoma cells by inhibiting the expression of MAOB.
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