Effects Of Silencing CPNE1,FUNDC2,KCTD14 And Vimentin Genes On Proliferation Of MDA-MB-231 Cells

Objective To explore the effects of CPNE1,FUNDC2,KCTD14 and Vimentin genes in silencing MDA-MB-231 cells and their effects on proliferation,initially provide a theoretical basis for the prevention and treatment of triple-negative breast cancer.Methods1.Firstly,three RNA silencing targets were designed for 20 genes to be tested,and three plasmids with different targets were mixed or packaged separately.MDA-MB-231 cells were infected by lentivirus packaging vector.Then,the cells were photographed and automatically counted by Celigo,and the effects of target gene on cell proliferation were compared,and the genes with obvious proliferation inhibition phenotype were initially screened.2.The mRNA expression levels of CPNE1,FUNDC2,KCTD14 and VIM genes were silenced by qRT-PCR.3.The effects of CPNE1,FUNDC2,KCTD14 and VIM genes on the short-term proliferation of cells were detected by MTS assay.4.The effects of CPNE1,FUNDC2,KCTD14 and VIM genes on the long-term proliferation of cells were detected by plate colony formation assay.Results1.The results of high content screening showed that among the 20 genes to be tested,the genes with obvious inhibition of phenotype were CPNE1,Vimentin,KCTD14,and FUNDC2.2.qRT-PCR showed that the expression levels of mRNA in the CPNE1,FUNDC2,KCTD14,and VIM groups were significantly lower than those in the negative control group(NC group)(P<0.001).3.MTS results showed that the short-term proliferation ability of cells in the CPNE1,FUNDC2,KCTD14,and VIM groups was significantly weakened,and the short-term proliferation ability of the CPNE1 experimental group was the most significant(P<0.001).4.The plate colony formation test showed that the long-term proliferation ability of the cells in the CPNE1,FUNDC2,KCTD14 and VIM groups was significantly weakened,and the long-term value-adding ability of the CPNE1 experimental group was the most significant(P<0.001).Conclusions1.High-content screening results showed that the genes with obvious proliferation inhibition phenotype were: CPNE1,VIM,KCTD14,and FUNDC2.2.The lentiviral-packaged CPNE1,FUNDC2,KCTD14,and VIM gene vectors can effectively silence the expression of the target gene of MDA-MB-231 cells.3.The silencing of CPNE1,FUNDC2,KCTD14,and VIM genes significantly inhibited the proliferation of MDA-MB-231 cells,with the most significant inhibition after CPNE1 gene silencing.