| Objectives:（1）To investigate the association between MIF gene promoter region-173G/C（rs755622）polymorphism and coronary artery disease（CAD）by systematic review and meta-analysis;（2）To investigate the association between MIF gene polymorphisms（rs2070766,rs755622,rs5844572）and Acute coronary syndrome（ACS）and the risk factors for ACS,and to construct a prediction model for ACS.The predictive performance of the nomogram was evaluated and verified from three aspects:discrimination,calibration and clinical applicability.Through follow-up,the incidence of Major adverse cardiovascular events（MACE）in different genotypes of MIF gene（rs2070766,rs755622,rs5844572）polymorphism was analyzed.（3）Dual-luciferase reporter assay and electrophoretic mobility shift assay were used to investigate the possible pathogenic mechanism of MIF gene promoter mutations.Methods:（1）Using"MIF","gene polymorphism","single nucleotide polymorphism"and"coronary heart disease"as key words,In ten thousand data knowledge service platform,China hownet pool,d Pu Zhong Wen periodical service platform,Chinese science citation database,the library of evidence-based medicine,biomedical literature retrieval system（Pub Med）,biological medicine and pharmacology abstract database（Embase）,and other Chinese and foreign biomedical database retrieval published in February,2021,31 before about M Literature on the association between IF rs755622 polymorphism and CHD.（2）In this case-control study,three polymorphisms of MIF gene（rs2070766,rs755622 and rs5844572）were selected and genotyped by SNPscanTM multiple single nucleotide polymorphism typing kit.The association between MIF gene polymorphism and ACS was analyzed.The predictors were selected based on multivariate Logistic regression analysis and LASSO regression method.The selected variables were included in R software to construct a nomogram model in the form of R software to predict the probability of ACS,and the nomogram model was internally validated and evaluated.To further clarify the effect of susceptibility genes on the long-term prognosis of ACS patients,survival curves were used to explore the incidence of MACE in different genotypes of three polymorphisms of MIF gene and whether the risk genotypes were related to the occurrence of MACE in ACS patients.Log-rank test was used to compare the survival rate between groups.（3）The mutation site of MIF gene promoter sequence and the wild-type target gene fragment were constructed into p GL3-basic reporter gene vector,and the constructed reporter gene vector and the internal control plasmid were co-transfected into293T tool cells by liposome.The relative luciferase activity of MIF gene promoter was analyzed by dual fluorescence reporter gene analyzer.Changes in the transcriptional activity of gene promoters caused by genetic mutations were detected.Conventional gel electrophoretic mobility shift assay and competitive gel electrophoretic mobility shift assay were also performed to verify whether MIF gene promoter region could bind to nuclear protein transcription factor and whether the binding was specific.Stata Corp Stata12.0,SPSS 22.0,R statistical software and Graph Pad Prism 8 were used for statistical analysis.Epidata 3.0 software was used for clinical data entry and check management.Results:（1）In general,the MIF-173 G/C（rs755622）C allele carriers had an increased risk of CAD.Compared with the wild type GG genotype,the mutant CC genotype carriers had an increased risk of CAD.Subgroup analysis showed that CC genotype carriers had an increased risk of CHD in Asian population,while CC genotype and GC genotype carriers had an increased risk of CHD in Caucasian population.（2）The frequency of GG genotype of MIF rs2070766 was higher in ACS patients than in controls（6.2%vs.3.8%,P=0.034）.Multivariate regression analysis showed that GG genotype of MIF rs2070766was associated with the risk of ACS.The GG genotype had a higher risk of ACS（OR=1.739,95%CI:1.022-2.962,P=0.042）.Based on multivariate Logistic regression analysis,the predictive factors of ACS were screened,and a nomogram model for predicting the risk of ACS was constructed.The prediction model had good discrimination.The self-sampling method was repeated 1000 times,and the Area under curve（AUC）was0.784(95%CI:The calibration curve showed good prediction accuracy and Hosmer-Lemeshow test P=0.515.The DCA curve showed that when the threshold probability of ACS was between 0.30 and 0.95,the nomogram model could produce better net clinical benefit in identifying patients at risk of ACS.（3）The frequency of CC genotype of MIF rs755622 was significantly higher in ACS patients than in controls（5.69%vs.2.71%,P=0.005）.Multivariate regression analysis showed that CC genotype was associated with the risk of ACS.CC genotype had a higher risk of ACS（OR=2.006,95%CI:1.158-3.567,P=0.015）.Based on multivariate Logistic regression analysis and LASSO regression analysis,the predictive factors of ACS were screened to construct a nomogram model for predicting the risk of ACS.The prediction model had good discrimination.The self-sampling method was repeated for 1000 times,and the AUC was0.790(95%CI:The calibration curve indicated that the nomogram had good prediction accuracy and Hosmer-Lemeshow test P=0.216.The DCA curve showed that when the threshold probability of ACS was greater than 20%,the net clinical benefit level of the nomogram was higher.（4）The CATT7/7 genotype of MIF rs5844572 was associated with a higher risk of ACS than the CATT5/5genotype（OR=2.993,95%CI:1.411-6.703,P=0.006）.Based on multivariate Logistic regression analysis and LASSO regression analysis,the predictive factors of ACS were screened to construct a nomogram model for predicting the risk of ACS.The prediction model had good discrimination.The self-sampling method was repeated 1000 times,and the AUC was 0.791(95%CI:The calibration curve showed good prediction accuracy and Hosmer-Lemeshow test P=0.146.The DCA curve showed that when the threshold probability of ACS was greater than 18%,the net clinical benefit level of the nomogram was higher.（5）During a median follow-up of 25 months（range 12-60 months）,ACS patients with GG genotype of MIF rs2070766had a higher risk of MACE events than those with CC or CG genotype.However,no positive results were found for MIF rs755622 and rs5844572.（6）Dual-luciferase reporter gene assay suggested that the mutations of rs755622 and rs5844572 in the promoter region of MIF gene could enhance the transcriptional activity of MIF.Gel electrophoretic mobility shift assay and competitive gel electrophoretic mobility shift assay confirmed that the MIF gene promoter region specifically bound to nuclear protein.Conclusions:MIF rs755622 polymorphism is associated with susceptibility to CAD in Asian and Caucasian populations.MIF rs2070766,rs755622 and rs5844572 gene polymorphisms are associated with the susceptibility to ACS in Xinjiang.The nomogram model for predicting the risk of ACS has good discrimination,accuracy and clinical applicability.For MIF rs2070766,ACS patients with GG genotype have a higher risk of MACE events than those with CC and CG genotype.Mutations in the promoter region of MIF gene may affect the binding of MIF gene to transcription factors and regulate the transcription of gene,which may be related to the pathogenesis of ACS. |
