基于NF-κB p65信号通路探索PRMT5靶向调控RPA2对肺腺癌细胞活性和侵袭能力的影响OACSTPCD
Effect of PRMT5 targeting RPA2 on activity and invasion of lung adenocarcinoma cells based on NF-κB p65 signaling pathway
目的 基于NF-κB p65信号通路探索蛋白精氨酸甲基转移酶5(PRMT5)靶向调控复制蛋白A2(RPA2)对肺腺癌细胞活性和侵袭能力的影响.方法 选择肺腺癌细胞株A549、H1299,随机分为空白对照组、质粒对照组、RPA2高表达组、PRMT5沉默组、PRMT5沉默+RPA2高表达组.质粒对照组予阴性对照siRNA转染,RPA2高表达组予pcDNA3.1/RPA2转染,PRMT5沉默组予PRMT5 siRNA转染,PRMT5沉默+RPA2高表达组予PRMT5 siRNA和pcDNA3.1/RPA2转染,然后用嘌呤霉素筛选获得稳定转染细胞.空白对照组常规培养,不予转染.收集各组细胞,采用CCK-8法检测细胞活性,采用Transwell小室实验检测细胞侵袭能力,采用Western blotting法检测PRMT5、RPA2以及p-IκBα、p-NF-κB p65蛋白表达.结果 在A549细胞和H1299细胞中,RPA2高表达组PRMT5、RPA2蛋白表达及细胞活性、侵袭能力均高于空白对照组和质粒对照组,PRMT5沉默组PRMT5、RPA2蛋白表达及细胞活性、侵袭能力均低于空白对照组和质粒对照组(P均<0.05);PRMT5沉默组和PRMT5沉默+RPA2高表达组PRMT5、RPA2蛋白表达及细胞活性、侵袭能力均低于RPA2高表达组(P均<0.05);PRMT5沉默+RPA2高表达组PRMT5、RPA2蛋白表达及细胞活性、侵袭能力均高于PRMT5沉默组(P均<0.05).在A549细胞和H1299细胞中,RPA2高表达组p-IκBα、p-NF-κB p65蛋白表达均低于空白对照组和质粒对照组,PRMT5沉默组p-IκBα、p-NF-κB p65蛋白表达均高于空白对照组和质粒对照组(P均<0.05);PRMT5沉默组和PRMT5沉默+RPA2高表达组p-IκBα、p-NF-κB p65蛋白表达均高于RPA2高表达组(P均<0.05);PRMT5沉默+RPA2高表达组p-IκBα、p-NF-κB p65蛋白表达均低于PRMT5沉默组(P均<0.05).结论 PRMT5可通过靶向调控RPA2增强肺腺癌细胞活性和侵袭能力,其作用机制可能与激活NF-κB p65信号通路有关.
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.
辛春霞;李敏敏;董亮亮
青岛大学附属烟台毓璜顶医院肿瘤内科,山东烟台 264099
临床医学
肺腺癌蛋白精氨酸甲基转移酶5复制蛋白A2核因子κB p65信号通路
lung adenocarcinomaprotein arginine methyltransferase 5replication protein A2nuclear factor κB p65 signaling pathway
《山东医药》 2024 (016)
10-14 / 5
山东省医药卫生科技发展项目(2018WS025).
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