miR-141-3p靶向调控FOXA1对肺炎链球菌感染的肺泡上皮细胞增殖凋亡的影响

Effects of miR-141-3p targeting FOXA1 on proliferation and apoptosis of alveolar epithelial cells infected by Streptococcus pneumoniae

目的探讨miR-141-3p对肺炎链球菌感染的肺泡上皮细胞增殖及凋亡的影响及其分子机制。方法以AECⅡ细胞为研究对象,分别将miR-NC、miR-141-3p mimics、si-NC、si-FOXA1、miR-141-3p mimics+pcDNA3.1、miR-141-3p mimics+pcDNA3.1-FOXA1转染至AECⅡ细胞,分别记作第1,2,3,4,5,6转染组,同时将且未经处理的细胞作为对照组、肺炎链球菌培养细胞作为模型组。实时荧光定量聚合酶链反应(qRT-PCR)检测细胞中miR-141-3p的表达水平;蛋白质印迹法检测FOXA1的表达量;甲基噻唑基四唑(MTT)与流式细胞术分别检测细胞增殖及凋亡。结果对照组、模型组、第1,2,3,4,5,6转染组2 h细胞存活率分别为(100.00±5.18)%,(47.19±3.14)%,(47.28±3.15)%,(86.73±4.63)%,(47.38±3.25)%,(79.93±4.23)%,(86.75±4.51)%,(57.90±3.34)%;细胞凋亡率分别为(8.11±1.12)%,(26.23±1.83)%,(26.23±2.83)%,(10.37±1.51)%,(26.23±2.83)%,(13.36±1.57)%,(10.35±1.57)%,(18.91±1.86)%。对照组与模型组比较,第1转染组与第2转染组比较,第3转染组与第4转染组比较,第5转染组与第6转染组比较,差异均有统计学意义(P<0.05)。结论miR-141-3p过表达可通过抑制FOXA1表达而抑制肺炎链球菌诱导的肺上皮细胞凋亡,促进细胞增殖。

Objective To investigate the effect of miR-141-3p on proliferation and apoptosis of alveolar epithelial cells infected with pneumococcal infection and its molecular mechanism.Methods Using AECⅡcells as the research object,miR-NC,miR-141-3p mimics,si-NC,si-FOXA1,miR-141-3p mimics and pcDNA3.1,miR-141-3p mimics and pcDNA3.1-FOXA1 were transfected into AECⅡcells and named as transfection 1 group,transfection 2 group,transfection 3 group,transfection 4 group,transfection 5 group,transfection 6 group,while untreated cells were assigned to control group,and Streptococcus pneumoniae culture cells were assigned to model group.quantitative reverse transcriptase polymerase chain reaction(qRT-PCR)was used to detect miR-141-3p expression in cells.The expression of FOXA1 was detected by Western blot.MTT and flow cytometry were used to detect cell proliferation and apoptosis,respectively.Results In the control group,model group,transfection 1,2,3,4,5,6 group,the 2 h cell survival rates were(100.00±5.18)%,(47.19±3.14)%,(47.28±3.15)%,(86.73±4.63)%,(47.38±3.25)%,(79.93±4.23)%,(86.75±4.51)%,(57.90±3.34)%;the cell apoptosis rates were(8.11±1.12)%,(26.23±1.83)%,(26.23±2.83)%,(10.37±1.51)%,(26.23±2.83)%,(13.36±1.57)%,(10.35±1.57)%,(18.91±1.86)%.Comparing the control group with the model group,the transfection 1 group was compared with the transfection 2 group,the transfection 3 group was compared with the transfection 4 group,and the transfection 5 group was compared with the transfection 6 group.The differences were statistically significant(P<0.05).Conclusion Overexpression of miR-141-3p inhibits the apoptosis of lung epithelial cells induced by Streptococcus pneumoniae by inhibiting the expression of FOXA1 and promotes cell proliferation.