Molecular Mechanism Of PRDX2 Gene Silencing Inhibits Invasion And Metastasis Of Breast Cancer Cells

Objective:The present study endeavors to examine the effect of PRDX2,a gene that is overexpressed in triple negative breast cancer cells,on cell migration and invasion.We aim to elucidate the specific mechanism by which PRDX2 promotes these traits and thereby provide innovative ideas and theoretical support for targeting PRDX2 as a therapeutic strategy for triple negative breast cancer.Methods:The present investigation utilized network database analysis to compare PRDX2 differential expression between normal and tumor tissues.PRDX2 lentivirus-based interference sequences were constructed and used to transfect cellular samples.RT-q PCR and Western Blotting were used to evaluate PRDX2 expression levels,and polyamine was employed to screen cellular samples to establish two stable low PRDX2-expressing triple-negative breast cancer cell lines(MDA-MB-231,MDA-MB-468).The scratch and Transwell methods were used to evaluate the migration and invasion capacities of the PRDX2 stable and low-expressing cell lines.Western blotting was then used to detect the expression levels of E-cadherin and N-cadherin proteins,indicators of the epithelial-mesenchymal transition(EMT).RNA sequencing was conducted to observe changes in gene expression in MDA-MB-231 cells with stably low PRDX2 gene expression,and gene changes were compared with those in a control group.The migrating and invading genes were then intersected with the migration-related and invasion-related genes,and the relevant genes(FN1,COL6A2,ITGB3)were discovered through enrichment analysis of the KEGG and GO pathways.RT-q PCR and Western Blotting confirmed the expression levels of the three related genes in the stable and low-expressing PRDX2 genome compared to the control group to verify the RNA sequencing results.Furthermore,si RNA was used to knock down the three genes(FN1,COL6A2,and ITGB3)in MDA-MB-231 cells with low PRDX2 expression to investigate the changes in migration and invasion.Transcriptional factors of invasion and migration genes selected via the intersection of RNA sequencing outcomes and the GESA database were analyzed and screened further in the TRRUST database,followed by Western Blotting for extended verification.Results:Bioinformatics analysis of breast cancer indicated high expression of PRDX2.Scratch and Transwell tests showed that the migration and invasion of the stably low expression PRDX2 triple-negative breast cancer cell line significantly decreased compared to the control group.Evaluation of the epithelial-mesenchymal transition(EMT)indexes via Western blotting revealed an increase in E-cadherin protein level and a decrease in N-cadherin protein level.FN1,COL6A2,and ITGB3 genes were selectively knocked down through reverse transcription RNA.RT-q PCR and Western blotting revealed decreased expression levels of MMP2,MMP9,and N-cadherin,and increased expression level of E-cadherin.The scratch test results showed that the migration of the MDA-MB-231 cell line with stable and low expression of the PRDX2 gene decreased after the respective knockdown of the three genes,with FN1 knocking down resulting in the most significant decrease.Further analysis via the TRRUST database and Western blotting confirmed that SP1 is the co-transcription factor for the aforementioned genes.Conclusion:This investigation detected high expression of the PRDX2 gene in breast cancer.Notably,triple-negative breast cancer cells with stable low expression of PRDX2 demonstrated reduced migration and invasion abilities.Regulating the transcription factor SP1 via PRDX2 affects downstream changes in the FN1 gene,thus raising the migration and invasion ability of breast cancer.As such,targeting the PRDX2 gene may prove to be a novel strategy for treating breast cancer.Figure [ 6 ] Table[ 5 ] Reference [100]...