Expression of miR-34a-5p in serum of patients with chronic obstructive pulmonary disease(COPD)and its effect on proliferation and apoptosis of smooth muscle cells and regulation of catenin beta 1(CTNNB1)
Objective To investigate the expression of miR-34a-5p in the serum of COPD patients and its regulation on smooth muscle cell proliferation,apoptosis and catenin beta 1(CTNNB1).Methods Select 50 stable patients with chronic obstructive pulmonary disease(COPD)who visited the hospital from Janu-ary to December 2023(COPD group),and another 50 healthy individuals who underwent physical exami-nations at physical examination center during the same period(normal group).qRT-PCR was used to de-tect the levels of miR-34a-5p and CTNNB1 mRNA in serum,Person correlation analysis was performed to determine the correlation between miR-34a-5p and CTNNB1 mRNA,and ROC curve analysis was per-formed to evaluate the diagnostic value of miR-34a-5p and CTNNB1 mRNA.5 of the 30 rats were random-ly selected as blank group,and the other 25 rats were used to establish the rat model of COPD,extract rat ASMCs,and divided ASMCs into blank group(normal rat ASMCs),model group(chronic obstructive pulmonary disease rat ASMCs),miR-34a-5p-NC group,miR-34a-5p mimics group,miR-34a-5p siRNA group,si-CTNNB1 group,miR-34a-5p mimics+si-CTNNB1 group and miR-34a-5p siRNA+si-CTNNB1 group.Cell proliferation rate was detected by CCK-8,cell apoptosis was detected by flow cytometry,CT-NNB1,β-catenin,Wnt1 and GSK3β levels were detected by Western blot,and the targeting relationship between miR-34a-5p and CTNNB1 was verified by dual luciferase reporter gene.Results The expression of miR-34a-5p in COPD group was significantly lower than that in normal group,and the expression of CTNNB1 mRNA was higher than that in normal group(P＜0.05).miR-34a-5p was negatively correlated with CTNNB1 mRNA(r=-0.551,P＜0.001).The AUC value of miR-34a-5p in the diagnosis of COPD was 0.912(95％CI:0.851～0.974),The AUC value of CTNNB1 mRNA in the diagnosis of COPD was 0.832(95％CI:0.775～0.909).Compared with the blank group,the proliferation ability,CTNNB1 andβ-catenin protein expression of ASMCs in the model group were increased,while the apoptosis rate,Wnt1 and GSK3β protein expression were decreased(P＜0.05).Compared with the model group,the prolifera-tion capacity of ASMCs,the expression of CTNNB1 and β-catenin protein in miR-34a-5p mimics group and si-CTNNB1 group were decreased,and the apoptosis rate,the expression of Wnt1 and GSK3β protein were increased(P＜0.05),while the miR-34a-5p siRNA group showed the opposite trend.Compared with miR-34a-5p mimics group,the proliferation capacity of ASMCs,the expression of CTNNB1 and β-catenin protein in miR-34a-5p mimics+si-CTNNB1 group were decreased,and the apoptosis rate and the expres-sion of Wnt1 and GSK3β protein were increased(P＜0.05).Compared with miR-34a-5p siRNA group,the proliferation capacity of ASMCs,the expression of CTNNB1 and β-catenin protein were decreased,and the apoptosis rate and the expression of Wnt1 and GSK3β protein were increased in miR-34a-5p siRNA+si-CTNNB1 group(P＜0.05).The results of dual luciferase assay showed that overexpression of miR-34a-5p could decrease the luciferase activity of CTNNB1-WT(P＜0.05).Conclusion The expression of miR-34a-5p in serum of the patients with COPD was significantly decreased,and it was negatively correlated with CTNNB1,which is helpful for evaluating the occurrence of COPD.Moreover,miR-34a-5p can regulate the proliferation and apoptosis of ASMCs,and it may be related to the targeted modulation of CTNNB1 expression.
万方数据
维普
