Effect of LncRNA MCM3AP-AS1 on Malignant Progression of Breast Cancer by Regulating miR-424-5p/PSAT1 Axis
Objective:To investigate the effect of long non-coding RNA(lncRNA)microchromosome maintenance protein 3-as-sociated protein-antisense chain 1(MCM3AP-AS1)on malignant progression of breast cancer by regulating microribonucleic acid(miR)-424-5p/phosphoserine aminotransferase 1(PSAT1)axis.Methods:The expression levels of LncRNA MCM3AP-AS1,miR-424-5p and PSAT1 mRNA in human normal breast epithelial cells MCF-10A and breast cancer cell lines MDA-MB-231,BT549 and BT20 were de-tected by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).LncRNA MCM3AP-AS1 low expression model,miR-424-5p knockdown and overexpression model were constructed,and the success of transfection model construction was verified by RT-qPCR method.The proliferation,migration and invasion of breast cancer cell lines were detected by methyl thiazolyl tetrazolium(MTT)assay and Transwell assay respectively.The expressions of cyclin d1(CyclinD1),E-cadherin(E-cadherin),N-cadherin(N-cad-herin)and PSAT1 proteins in MDA-MB-231 cells were detected by Western blotting.After bioinformatics analysis,the targeting rela-tionship between LncRNA MCM3 AP-AS1 and miR-424-5p,miR-424-5p and PSAT1 was verified by double luciferase reporter gene as-say and RNA immunoprecipitation(RIP)assay respectively.Results:The expression levels of MCM3AP-AS1 and PSAT1 mRNA in breast cancer cell lines MDA-MB-231,BT549 and BT20 were significantly higher than those in MCF-10A(P＜0.05),and the expression level of miR-424-5p was significantly lower than that in MCF-10A(P＜0.05).Knockdown of MCM3AP-AS1 or overexpression of miR-424-5p could reduce the absorbance(OD490)value of MDA-MB-231 cells,the number of cell migration and cell invasion,the ex-pression levels of CyclinD1,N-cadherin and PSAT1 proteins(P＜0.05),and increase the expression level of E-cadherin protein(P＜0.05).Knockdown of miR-424-5p reversed the effects of down-regulation of MCM3AP-AS1 on the proliferation,migration and invasion of MDA-MB-231 cells and the expression of related proteins.The dual luciferase reporter gene assay and RIP assay confirm that MCM3AP-AS1 targeted and negatively regulated the expression of miR-424-5p,and miR-424-5p targeted and negatively regulated the expression of PSAT1.Conclusion:LncRNA MCM3AP-AS1 is highly express in breast cancer,and its low expression can inhibit the ma-lignant progression of breast cancer by targeting the miR-424-5p/PSAT1 axis.
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