Preliminary Experimental Study On Circ-MTDH/miR-22 Axis Regulation Of Osteosarcoma Cell Proliferation And Apoptosis

Objective The development of osteosarcoma is considered to be closely related to circular RNAs and micro RNAs.Previously,researchers reported that Micro RNA-22 could target and bind to the MTDH in osteosarcoma to inhibit autophagy and affect chemotherapy sensitivity of osteosarcoma.Other scholars have found that circ MTDH inhibits the proliferation and invasion of non-small cell lung cancer by combining with miR-630.However,there is a lack of relevant reports on the role of miR-22 on circ-MTDH and its relationship with osteosarcoma cell apoptosis.The purpose of this experiment was to study the expression of Circ MTDH in osteosarcoma cell lines,to verify the sponge effect of Circ MTDH and Micro RNA-22,and to further study the relationship between Micro RNA-22 and osteosarcoma cell proliferation and apoptosis.To preliminarily explore the mechanism of Circ-MTDH/miR-22 axis in osteosarcoma.Methods In this study,the expression levels of Circ MTDH,miR-22 and MTDH in osteosarcoma cell lines and h FOB1.19 were first detected by q RT-PCR.Gene database,luciferase assay were employed to predict and validate the targeting relationships between Circ MTDH and miR-22.In vitro cell experiments,osteosarcoma cell line(MG63)was divided into miR-22 NC control group and miR-22 mimic transfection group by lipofection,and the transfection efficiency of miR-22 was detected.Proliferation and apoptosis of MG63 cells in NC control group and Mir-22 mimic group were detected by MTT assay and flow cytometry,and bcl-2,Bax,Cleaved caspase-3 were detected by PCR and Western blotting.Finally,statistical methods were used to analyze whether the differential expression of Circ-MTDH was statistically different and the correlation between miR-22 and the above detection indicators.Results Circ MTDH and MTDH were significantly up-regulated in osteosarcoma cell lines,of which circ-0085005 was the most prominent,while miR-22 was significantly down-regulated.Circ-MTDH is mainly located in the cytoplasm of osteosarcoma cells.The results of gene database review and luciferase assay showed that Circ MTDH has a targeted binding site for miR-22,which can bind to the miR-22.The results of MTT assay showed that miR-22 could inhibit osteosarcoma cell’s proliferation activity.Upregulation of miR-22 can increase caspase-3 and Bax and down-regulate bcl-2 expression in osteosarcoma cells.Based on this,it can be preliminarily inferred that Circ MTDH promotes apoptosis by competitively binding to miR-22.Conclusion Overexpression of miR-22 in osteosarcoma MG63 cells can promote apoptosis.Circ-MTDH can sponge-binding miR-22 and may promote osteosarcoma cell proliferation and inhibit apoptosis through this regulatory axis.It is a potential molecular marker and possible therapeutic target for osteosarcoma,but further experiments are needed to confirm it.The result is not yet certain.