Association Between MAPK1 And PRKCA Gene Polymorphisms In MAPK Signaling Pathway And The Susceptibility Of Ischemic Stroke And Coronary Artery Disease

Objective:Previous studies suggested that the MAPK1 gene and PRKCA gene of MAPK signaling pathway may be involved in the pathological mechanisms of ischemic stroke?IS?and coronary artery disease?CAD?.This study aimed to explore the association between genetic polymorphisms in the3'-UTR region of MAPK1 and PRKCA genes and the risk of IS or CAD,as well as the IS or CAD related clinical parameters.We then used the dual luciferase reporter assay to explore the effects of 3'-UTR polymorphism on the targeted binding of miRNA to target gene,so that providing molecular genetic basis for the interpretion of the genetic mechanisms of these two diseases and the exploration of their therapeutic targets.Methods:A total of 550 patients of ischemic stroke,550 patients of coronary artery disease,and 550 controls were included in this study.The relative expression levels of MAPK1 and PRKCA mRNA in peripheral blood of the subjects were determined by quantitative real-time polymerase chain reaction?qRT-PCR?.The genotyping of MAPK1 gene rs6928,rs13515,rs1063311 polymorphisms and PRKCA gene rs2286674,rs7342847polymorphisms was performed using Massarray SNP genotyping assay.The dual luciferase reporter assay was used to explore the effect of MAPK1 gene rs13515 polymorphism on the targeted binding of hsa-miR-187 to MAPK1 gene.Results:1.Comparison of MAPK1 and PRKCA gene mRNA expression levels between case group and control group:The expression level of MAPK1mRNA in IS group was higher than that in control group?P=0.045?,while the expression level of PRKCA gene mRNA in IS group was lower than that in control group?P<0.001?.The mRNA expression level of MAPK1 gene in CAD group was higher than that in control group?P<0.001?,while the mRNA expression level of PRKCA gene in CAD group was lower than that in control group?P=0.030?.2.Association analysis of MAPK1 and PRKCA gene polymorphisms and the risk of ischemic stroke or coronary artery disease:?1?MAPK1 gene rs13515 polymorphism was significantly associated with the risk of IS(additive model:OR_adj=1.26,95%CI=1.03-1.55,P_adj=0.028;dominant model:OR_adj=1.40,95%CI=1.10-1.78,P_adj=0.007).?2?PRKCA gene rs2286674polymorphism(additive model:OR_adj=0.83,95%CI=0.70-0.98,P_adj=0.028;dominant model:OR_adj=0.74,95%CI=0.57-0.95,P_adj=0.017)and rs7342847polymorphism(additive model:OR_adj=1.28,95%CI=1.08-1.52,P_adj=0.004;dominant model:OR_adj=1.43,95%CI=1.12-1.83,P_adj=0.004)were both significantly associated with the CAD risk.3.Association analysis of MAPK1,PRKCA gene polymorphisms with clinically relevant quantitative traits of ischemic stroke or coronary artery disease:?1?In terms of blood pressure levels,MAPK1 gene rs1063311 polymorphism(additive model:P_adj=0.021;recessive model:P_adj=0.013)and the PRKCA gene rs2286674 polymorphism(recessive model:P_adj=0.039)were both found to be associated with the SBP levels in IS patients.?2?In terms of blood glucose levels,PRKCA gene rs7342847polymorphism was significantly associated with P2hPG levels in CAD patients(dominant model:P_adj=0.033).?3?In terms of blood lipid levels,PRKCA gene rs7342847 polymorphism was significantly associated with LDL-C levels in IS patients(additive model:P_adj=0.023;recessive model:P_adj=0.015),rs13515polymorphism of MAPK1 gene was significantly associated with APO-A1 levels in CAD patients(dominant model:P_adj=0.046),and rs1063311 polymorphism was significantly associated with HDL-C levels in CAD patients(dominant model:P_adj=0.049),PRKCA gene rs2286674 polymorphism was significantly associated with APO-B levels in CAD patients(recessive model:P_adj=0.020).?4?In terms of coagulation function levels,the rs13515 polymorphism was not only significantly associated with APTT levels in IS patients(recessive model:P_adj=0.003),but also associated with PT levels in CAD patients(recessive model:P_adj=0.006),rs7342847 polymorphism was not only significantly associated with FIB levels in IS patients(recessive model:P_adj=0.010),but also associated with PTA levels in CAD patients(additive model:P_adj=0.021;dominant model:P_adj=0.045).?5?In addition,rs6928(recessive model:P_adj=0.043)and rs13515(additive model:P_adj=0.028;recessive model:P_adj=0.000)polymorphisms were both significantly associated with CRP levels in CAD patients.4.Dual luciferase reporter assay:hsa-miR-187 can bind to the rs13515 wild type?C allele?of the 3'-UTR region of MAPK1 gene,and the rs13515 mutant?T allele?can alter the binding of hsa-miR-187 to the rs13515 of MAPK1 gene.Conclusions:1.The mRNA expression of MAPK1 gene and PRKCA gene may be related to the occurrence of ischemic stroke and coronary artery disease,MAPK1 gene and PRKCA gene may affect the incidence of ischemic stroke and coronary artery disease.2.MAPK1 gene rs13515 polymorphism was associated with the risk of ischemic stroke,and the PRKCA gene rs2286674 and rs7342847 polymorphisms were both associated with the risk of coronary artery disease.3.Genetic polymorphisms of MAPK1 and PRKCA genes may affect blood pressure and coagulation function in ischemic stroke,and PRKCA gene polymorphisms may be involved in lipid metabolism in ischemic stroke.Genetic polymorphisms of MAPK1 and PRKCA genes may affect the lipid metabolism and coagulation function of coronary artery disease,and MAPK1 polymorphism may also be involved in the inflammatory response of coronary artery disease,PRKCA gene polymorphism may also affect the blood glucose metabolism of coronary artery disease.4.hsa-miR-187 may regulate the expression level of MAPK1 gene by binding to MAPK1 gene,and this binding may be altered by rs13515 polymorphism,which may affect the occurrence of ischemic stroke.