Effects of Midazolam on Proliferation Apoptosis and Invasion of Osteosarcoma Cells by Regulating the YAP/TAZ Signaling Pathway
Objective:To investigate the effects of midazolam(MDZ)on the proliferation,apoptosis and invasion of osteosarcoma(OS)cells by regulating the Yes associated protein(YAP)/PDZ transcriptional activator of transcription(TAZ)signaling pathway.Methods:qRT-PCR and western blot were applied to de-tect the expression of YAP,TAZ mRNA,and proteins in normal osteoblast and OS cell lines,and to screen the optimal intervention cell line.OS cells were intervened with different concentrations of MDZ(0,12.5,25,50,100,200 μmoL/L),MTT method was applied to detect cell proliferation activity and screen the opti-mal intervention concentration;OS cells were randomly divided into control group,MDZ group,pcDNA3.1 group,and pcDNA3.1-YAP/TAZ group,and transfection efficiency was detected using qRT-PCR method;EdU staining,flow cytometry,and Transwell cells were applied to detect cell proliferation,apoptosis,and in-vasion,respectively;Western blot method was applied to detect the expression of proliferating cell nuclear an-tigen(PCNA),Bcl-2 associated X protein(Bax),N-cadherin,E-cadherin,and YAP,TAZ proteins;OS nude mouse model was constructed,and grouped into a control group and an MDZ group,immunohistochemi-cal methods were applied to detect the expression of Ki-67,YAP,and TAZ proteins in transplanted tumor tis-sue,and TUNEL staining was applied to detect apoptosis.Results:The expression levels of YAP and TAZ mRNA and protein in OS cell lines were obviously increased(P<0.05);MDZ obviously reduced the expres-sion of PCNA,N-cadherin,YAP,and TAZ,inhibitd the proliferation and invasion of MG63 and U20S cells,increased the expression of Bax and E-cadherin,and promoted cell apoptosis.Overexpression of YAP/TAZ was able to reverse the inhibitory effects of MDZ on the proliferation and invasion of MG63 and U20S cells,and promote the apoptosis of MG63 and U20S cells(P<0.05);in vivo experiments showed that MDZ obvi-ously reduced the mass and volume of transplanted tumors,and the expression levels of Ki-67,YAP,and TAZ,promoted apoptosis of transplanted tumor cells(P<0.05).Conclusion:MDZ may inhibit the YAP/TAZ signaling pathway,inhibit the proliferation and invasion of OS cells,and promote apoptosis.
国家科技期刊平台
NSTL
万方数据
维普
