miR-17-5p靶向乙醛脱氢酶2减少肺炎链球菌诱导的人肺腺癌肺泡基底上皮细胞系A549损伤  

miR-17-5p targeting ALDH2 reduces the damage of human lung adenocarcinoma alveolar basal epithelial tissue cell line A549 induced by Streptococcus pneumoniae

作　　者：李维维 葛斌 邓涛 唐建军 骆财元 冯圣芳 毛羽 LI Wei-wei;GE Bin;DENG Tao;TANG Jian-jun;LUO Cai-yuan;FENG Sheng-fang;MAO Yu(Department of Pediatrics,Pidu District People s Hospital,Chengdu 611730,China;Department of Clinical Laboratory,Pidu District People s Hospital,Chengdu 611730,China)

机构地区：[1]成都市郫都区人民医院儿科,四川成都611730 [2]成都市郫都区人民医院检验科,四川成都611730

出　　处：《基础医学与临床》2020年第10期1362-1368,共7页Basic and Clinical Medicine

基　　金：四川省医学会科研课题(S16050)。

摘　　要：目的探讨miR-17-5p表达对肺炎链球菌(S.pneumoniae)诱导的人肺腺癌肺泡基底上皮细胞系A549损伤的影响及其作用机制。方法将A549细胞分为对照组、感染组(1×108 CFU/mL S.pneumoniae);anti-miR-17-5p干预组(S.pneumoniae感染前转染anti-miR-17-5p)、anti-miR-NC干预组(S.pneumoniae感染前转染anti-miR-NC)、pcDNA-乙醛脱氢酶2(ALDH2)干预组(S.pneumoniae感染前转染pcDNA-ALDH2)、pcDNA干预组(S.pneumoniae感染前转染pcDNA)。RT-qPCR检测细胞中miR-17-5p与ALDH2 mRNA水平,酶联免疫吸附剂法检测细胞培养上清中IL-6和IL-10水平,流式细胞计量术检测细胞凋亡,Western blot检测ALDH2、Bcl-2和Bax蛋白水平。双荧光素酶实验验证miR-17-5p与ALDH2之间靶向关系。结果与对照组比,感染组miR-17-5p水平显著升高(P<0.05);ALDH2 mRNA和蛋白水平显著降低(P<0.05);IL-10和Bcl-2水平显著降低(P<0.05);凋亡率、IL-6和Bax水平显著升高(P<0.05)。与anti-miR-NC干预组比,anti-miR-17-5p干预组IL-10和Bcl-2水平显著升高(P<0.05);凋亡率、IL-6和Bax水平显著降低(P<0.05)。与pcDNA干预组比,pcDNA-ALDH2干预组IL-10和Bcl-2水平显著升高(P<0.05);凋亡率、IL-6和Bax水平显著降低(P<0.05)。miR-17-5p负调控ALDH2表达,抑制ALDH2表达逆转了抑制miR-17-5p表达对S.pneumoniae诱导的A549细胞损伤。结论抑制miR-17-5p表达通过调控ALDH2降低S.pneumoniae诱导的人肺腺癌肺泡基底上皮细胞系A549损伤。Objective To investigate the effect of miR-17-5p expression on the injury of human lung adenocarcinoma alveolar basal epithelial cell line A549 induced by Streptococcus pneumonia and its mechanism.Methods A549 cells was divided into control group,infected group(1×108 CFU/mL S.pneumoniae),anti-miR-17-5p intervention group(transfected with anti-miR-17-5p before S.pneumoniae infection),anti-miR-NC intervention group(transfected with anti-miR-NC before S.pneumoniae infection),and pcDNA-ALDH2 intervention group(trans-fected with pcDNA-ALDH2 before S.pneumoniae infection),and pcDNA intervention group(transfected with pcDNA before S.pneumoniae infection).The level of miR-17-5p and ALDH2 mRNA was detected by RT-qPCR.IL-6 and IL-10 in cell culture supernatant were detected by enzyme-linked immunosorbent assay.Apoptosis was detected by flow cytometry.ALDH2,Bcl-2 and Bax protein were detected by Western blot.Dual luciferase assay verified the targeting relationship between miR-17-5p and ALDH2.Results Compared with the control group,the level of miR-17-5p in the infected group was significantly increased(P<0.05),while mRNA and the protein concentration of ALDH2,IL-10 and Bcl-2 were significantly decreased(P<0.05).However,apoptotic rate and the levels of IL-6 and Bax were significantly increased(P<0.05).Compared with the infected+anti-miR-NC group,IL-10 and Bcl-2 in the infected+anti-miR-17-5p group were significantly increased(P<0.05),the apoptotic rate,IL-6 and Bax were significantly decreased(P<0.05).Compared with the infected+pcDNA group,the levels of IL-10 and Bcl-2 in the infected+pcDNA-ALDH2 group were significantly increased(P<0.05),and the apoptotic rate and the levels of IL-6 and Bax were significantly decreased(P<0.05).miR-17-5p negatively regulated ALDH2 expression.Inhibition of ALDH2 expression reversed the effect of inhibiting miR-17-5p expression on S.pneumoniae-induced A549 cells damage.Conclusions Inhibition of miR-17-5p expression reduces S.pneumoniae-induced the damage of the cell line A549 by regulating A

关 键 词：miR-17-5p 乙醛脱氢酶2 肺炎链球菌 肺泡上皮细胞 细胞损伤 

分 类 号：R378[医药卫生—病原生物学]