Effect of PRMT5 targeting RPA2 on activity and invasion of lung adenocarcinoma cells based on NF-κB p65 signaling pathway
Objective To investigate the effect of protein arginine methyltransferase 5(PRMT5)targeting replica-tion protein A2(RPA2)on the activity and invasion of lung adenocarcinoma cells based on NF-κB p65 signaling pathway.Methods Lung adenocarcinoma cell lines A549 and H1299 were selected and randomly divided into the blank control group,plasmid control group,RPA2 high expression group,PRMT5 silencing group,PRMT5 silencing+RPA2 high expression group.Cells in the plasmid control group were transfected with negative control siRNA,RPA2 high expression group with pcDNA3.1/RPA2,PRMT5 silencing group with PRMT5 siRNA,and PRMT5 silencing+RPA2 high expres-sion group with PRMT5 siRNA and 10 μg pcDNA3.1/RPA2.Stable transfected cells were then screened with purinomy-cin.Cells in the blank control group were cultured without transfection.CCK-8 was used to detect cell activity,Transwell assay was used to detect cell invasion ability,and Western blotting was used to detect PRMT5,RPA2,p-IκBα,p-NF-κB p65 protein expression.Results In A549 cells and H1299 cells,the expression of PRMT5 and RPA2 protein,cell activ-ity and invasion ability of the RPA2 high expression group were higher than those of the blank control group and plasmid control group.PRMT5 and RPA2 protein expression,cell activity and invasion ability in the PRMT5 silencing group were lower than those in blank control group and plasmid control group(all P<0.05).PRMT5 and RPA2 protein expression,cell activity and invasion ability of the PRMT5 silencing group and PRMT5 silencing+RPA2 high expression group were lower than those of the RPA2 high expression group(all P<0.05).PRMT5 and RPA2 protein expression,cell activity and invasion ability in the PRMT5 silencing+RPA2 high expression group were higher than those in the PRMT5 silencing group(all P<0.05).In A549 cells and H1299 cells,the expression of p-IκBα and p-NF-κB p65 protein in the RPA2 high expression group was lower than that in the blank control group and plasmid control group,and the expression of p-IκBα and p-NF-κB p65 protein in the PRMT5 silencing group was higher than that in the blank control group and plasmid con-trol group(all P<0.05).The protein expression levels of p-IκBα and p-NF-κB p65 in the PRMT5 silencing group and PRMT5 silencing+RPA2 high expression group were higher than those in the RPA2 high expression group(all P<0.05).The protein expression levels of p-IκBα and p-NF-κB p65 in the PRMT5 silencing+RPA2 high expression group were lower than those in the PRMT5 silencing group(all P<0.05).Conclusion PRMT5 enhances the activity and invasion ability of lung adenocarcinoma cells by targeting RPA2,and its mechanism is related to the activation of NF-κB p65 signaling pathway.
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