Micro RNA-363-3p Regulates The Proliferation And Invasion Of Human Osteosarcoma Cells By Targeting PNO1

Background: Osteosarcoma(OS)is a worldwidely regarded as primary malignant bone tumor with a very high probability of metastasis and a very low survival rate of 5year.miR-363-3p is downregulated in a variety of malignant tumors and has a positive function in tumor inhibition.However,the role of it in osteosarcoma remains unclear yet.Objective: To research for the function of miR-363-3p in osteosarcoma.To provide bone tumors diagnosis and treatment with a new target.Methods: RT-q PCR was applied to check the relative level of miR-363-3p in two types of cell lines,as well as tissues;U2OS cells were infected by lentivirus with miR-363-3p overexpression vector and miR-363-3p negative control vector.MTT was arranged to test the proliferation ability of osteosarcoma cells successfully intecteed by lentivirus.Flow cytometry was used to analyze the changes of apoptosis ability of osteosarcoma cells in the two groups after successful infection.Flow cytometry was used to determine the number of different cycle cells.The expression level of apoptotic protein was detected by Western blotting.The apoptotic status of osteosarcoma cells after infection was observed by microscope,and then the apoptotic efficiency was evaluated..The expression of PNO1 in osteosarcoma tissues and infected U2 OS cells was detected by RT-q PCR and Western blotting.The dual luciferase gene assay was used to determine whether PNO1 was the target of miR-363-3p in osteosarcoma cells.Results: Compared with paracancer tissues and normal cell lines,miR-363-3p was down-regulated and PNO1 was up-regulated in osteosarcoma tissues and cell lines,showing a significant negative correlation between the two.The overexpression of miR-363-3p can limit the proliferation of osteosarcoma cells and accelerate the apoptosis of osteosarcoma cells.Western blotting analysis showed that U2 OS cells infected with miR-363-3p overexpression vector had a relatively lower PNO1 expression.A 3’-UTR double luciferase gene containing PNO1 messenger RNA was reported to indicate that PNO1 was a target of miR-363-3p.Conclusion: Overexpression of miRNA-363-3p can inhibit proliferation and promote apoptosis of osteosarcoma cells by targeting Pno1.