Effects Of Silencing CPNE1,KCTD14,FUNDC2 Genes On The Migration And Invasion Of MDA-MB-231 Cells

Objective To investigate target genes realted to migration,and to explore the effect of silencing target genes on MDA-MB-231 cells invasion.Provided a new therapeutic targets for triple-negative breast cancer.Methods1.3 RNA interference targets were designed for each of the 20 target genes.Three plasmids with different silencing sites were packaged with lentivirus to infect MDA-MB-231 cells in 2 ways,respectively and equally mix,called shRNA1,shRNA2,shRNA3 and shRNAmix.Negative control group(NC)was set.2.Using high-connotation screening scratch experiment(HCS),Celigo collected cell migration images,made data analysis to compare the callus ability,and selected the purpose genes related to cell migration.3.Western blot(WB)was used to detect the protein expression of the shRNA.4.The invasion ability of cells was tested by Transwell invasion test.5.The expression of epithelial-tissue-associated mesenchyme(EMT)proteins were detected by WB.ResultsThe HCS showed that the migration ability of CPNE1-shRNA,KCTD14-shRNA and FUNDC2-shRNA were obviously weakened than NC(Fold change>2).WB showed that the expression of corresponding proteins in CPNE1-shRNA1,KCTD14-shRNA1 and FUNDC2-shRNA2 group were significantly reduced compared with NC.The number of shRNA cells that pass through the transwell chambers were less than NC group.WB showed that the expressions of vimentin,snail and TCF8/ZEB1 in CPNE1-shRNA1,KCTD14-shRNA1 groups were decreased.The expressions of vimentin and TCF8/ZEB1 were decreased in FUNDC2-shRNA2 group.Conclusions1.Silencing CPNE1,KCTD14 and FUNDC2 genes inhibited the migration and invasion ability of MDA-MB-231 cells in triple-negative breast cancer.2.Silencing CPNE1,KCTD14 and FUNDC2 reduced the expression levels of EMT-related proteins in MDA-MB-231 cells.