微小RNA靶向胰岛再生源蛋白3A对肺上皮细胞增殖和凋亡的影响  

MicroRNA targeting pancreatic islet regeneration source protein 3A(REG3A)on the proliferation and apoptosis of lung epithelial cells

作　　者：王文涛[1] 郭健[2] 解敏[3] 郑志方[1] 王燕[1] WANG Wentao;GUO Jian;XIE min;ZHENG Zhifang;WANG Yan(Departemtn of Pediatrics,Affiliated Hospital Of Chengde Medical University,Chengde 067000,China;不详)

机构地区：[1]承德医学院附属医院小儿内科,河北承德067000 [2]承德医学院附属医院新生儿科,河北承德067000 [3]承德医学院图书馆,河北承德067000

出　　处：《实用医学杂志》2022年第4期439-445,共7页The Journal of Practical Medicine

基　　金：河北省科技支撑计划项目(编号:20200392)。

摘　　要：目的检测重症肺炎患儿血清中可能靶向抑制胰岛再生源蛋白3A(REG3A)的miRNA表达情况,并探究miR-137与miR-342-3p对肺上皮细胞增殖和凋亡的影响及机制。方法qRT-PCR检测重症肺炎患儿和健康儿童血清样本中相关miRNA和REG3A mRNA表达;将肺上皮细胞A549随机分为空白对照组、miR-137 NC组、miR-137 mimic组、miR-342-3p NC组、miR-342-3p mimic组,qRT-PCR检测miR-137、miR-342-3p和REG3A mRNA表达,Western blot检测REG3A、Caspase-3、Bcl-2、Bax和p53蛋白表达,双荧光素酶报告基因实验检测荧光活性,CCK-8法和EdU染色检测细胞增殖,流式细胞术检测细胞凋亡。结果重症肺炎患儿血清中miR-425-5p、miR-137、miR-325-3p、miR-342-3p、miR-203a-3p及miR-9-3p相对表达量均高于健康儿童,而REG3A mRNA相对表达量低于健康儿童(P<0.05);miR-137、miR-342-3p均与REG3A 3′-UTR存在碱基互补现象,并靶向调控其表达(P<0.05);miR-137过表达和miR-342-3p过表达均能够使A549细胞增殖活性和EdU阳性细胞率降低、细胞凋亡率升高,Caspase-3、Bax和p53蛋白表达增加而Bcl-2蛋白表达减少(P<0.05)。结论miR-137和miR-342-3p可能通过靶向REG3A抑制肺上皮细胞增殖,并促进细胞凋亡。Objective To detect the expression of miRNA in the serum of children with severe pneumonia,and to explore the effects and mechanisms of miR-137 and miR-342-3p on the proliferation and apoptosis of lung epithelial cells.Methods qRT-PCR was used to detect the expression of miRNA and REG3A mRNA in serum samples of children with severe pneumonia and healthy children;Lung epithelial cells A549 were randomly divided into blank control group,miR-137 NC group,miR-137 mimic group,miR-342-3p NC group,miR-342-3p mimic group,qRT-PCR detects miR-137,miR-342-3p and REG3A m RNA expression,Western Blot detects REG3A,Caspase-3,Bcl-2,Bax and p53 protein expression,the double luciferase reporter gene experiment detects fluorescence activity,CCK-8 method and EdU staining detected cell proliferation,flow cytometry detected cell apoptosis.Results The relative expression levels of miR-425-5p,miR-137,miR-325-3p,miR-342-3p,miR-203a-3p and miR-9-3p in the serum of children with severe pneumonia were higher than those of healthy children and the relative expression of REG3A mRNA was lower(P<0.05);Both miR-137 and miR-342-3p and REG3A3′-UTR have base complementation,and their expression is regulated by targeting(P<0.05);Both miR-137 overexpression and miR-342-3p overexpression can reduce the proliferation activity of A549 cells,reduce the rate of Ed U positive cells,increase the rate of apoptosis and the expression of Caspase-3,Bax and p53 proteins,while Bcl-2Protein expression decreased(P<0.05).Conclusion miR-137 and miR-342-3p may inhibit lung epithelial cell proliferation and promote cell apoptosis by targeting REG3A.

关 键 词：MIRNA 肺上皮细胞 胰岛再生源蛋白3A 增殖 凋亡 

分 类 号：R725.6[医药卫生—儿科]