The Roles And Mechanisms Of MiR-375 Participates In Wnt/?-catenin Pathway By Targeting FZD4 In The Progression Of Cervical Cancer

Objective:To investigate the roles of miR-375 on the function of proliferation,migration and invasion in cervical cancer cells and identify the target genes and pathway of miR-375 preliminarily,thus to explore the molecular mechanisms of miR-375 in cervical cancer progression.Methods:Using transient transfection technology,cervical cancer cell line SiHa was transfected with miR-375 mimics and inhibitor related suquences to construct the over-expression and low-expression cell models of miR-375,then transfected with FZD4 si RNA related suquences to construct the low-expression cell model of FZD4.Then using fluorescenece microscope to observe the transfection efficiency.The effect of transfection were further deternimed by Quantitative Real-time PCR and western blot for miR-375 and FZD4,respectively.After the construction of related cell model,western blot was used to detect the expression of ?-catenin protein in Wnt signaling.CCK8 method,wound healing test and transwell assay were used to detect the alterations of proliferation,migration and invasion,respectively.Results:1.Using cell transfection,the efficiency of each group containing miR-375 related suquence was over 90%,in miR-375 mimics group,the expression of miR-375 wasupregulated by(8.93±0.96)folds((49)=0.001),the ability of proliferation and migration was inhibited((49)=0.001),the ability of invasion was deregulated by(42.95±10.31)%((49)=0.046);in miR-375 inhibitor group,the expression of miR-375 was deregulated by(35.61±10.24)%((49)=0.001),the ability of proliferation and migration was enhanced((49)=0.001),the ability of invasion was upregulated by(67.68±13.66)%((49)=0.046),compared to responding negative control group;2.FZD4 was one of the potential target genes of mi R-375 by prediction,in miR-375 mimics group,the expression of FZD4 and ?-catenin proteins were deregulated((49)=0.023)((49)=0.001),and they were in highly positive correlated(R=0.951,(49)=0.001),moreover the expression of ?-catenin deregulated significantly when FZD4 was inhibited;3.Using cell transfection,the efficiency of each group containing FZD4 siRNA related suquence was over 90%,in FZD4 siRNA group,the expression of FZD4 protein was deregulated by(47.41±0.79)%((49)=0.001),the ability of proliferation and migration was inhibited((49)=0.001),the ability of invasion was deregulated by(48.32±7.96)%((49)=0.046),compared to responding negative control group.Conclusion:1.Low-expression of miR-375 in cervical cancer cell enhances cell proliferation,migration and invasion.It implies miR-375 may participate the progression of cervical cancer;2.MiR-375 participates in Wnt/?-catenin signaling by targeting FZD4,thus to achieve the regulation of biological functions of cervical cancer cells.