The Role And Significance Of AURKA In Aortic Dissecting Aneurysm

Objectives To analyze the relationship between the expression of Aurora kinase A(AURKA)in the tissues of patients with aortic-dissecting aneurysm(ADA)and its basic clinical data and biochemical indicators.To explore the role and significance of AURKA in ADA and normal vascular tissues.To determine whether AURKA regulates the proliferation and migration of vascular smooth muscle cells(VSMCs)through the GSK-3β/β-catenin signaling pathway.Methods Select tissue samples from 43 ADA patients who were diagnosed and treated in the Fourth Hospital of Hebei Medical University from January 2016 to June 2019.All samples were confirmed by two experienced pathologists.The age of the patient was 22 to74 years old.They were divided into ADA group(n=31)and ADA rupture(RADA)group(n=12).Non-ADA aortic wall tissue specimens were obtained from organ donors as controls(n=11).The pathological changes of the tissue samples were observed by hematoxylin and eosin(HE)staining.Real-time quantitative polymerase chain reaction(RT-q PCR)was used to detect the expression of AURKA mRNA in normal blood vessel tissues,ADA tissues and ruptured ADA tissues.Western blot was used to detect the expression of AURKA protein in the above-mentioned aortic tissues.Immunohistochemical staining was used to detect the expression of AURKA protein in the above-mentioned aortic tissues.3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)was used to detect the effect of AURKA on VSMC proliferation.Scratch healing test to detect the effect of AURKA on the migration of VSMC.Analysis of AURKA expression in ADA was divided into low expression group and high expression group,and compared with basic clinical data and biochemical indicators.Results 1 A total of 61 samples were collected for the baseline data of this study.Excluding 7 cases with incomplete data,54 cases can be analyzed.The age range is 22 to74 years old.Of the 54 patients in the study,37 were male,accounting for 68.5%.There are 17 women,accounting for 31.5%.Among them,35 have retired,accounting for 64.8%.16 people with overweight body mass index,accounting for 26.0%.45 people with a history of hypertension,accounting for as high as 83.3%.The majority of the study subjects have a history of drinking,accounting for 75.9%.2 The structure of the blood vessel wall of the control group is clear and the VSMC are arranged neatly.The blood vessel walls in the ADA group and RADA group were significantly thickened.The arrangement of VSMCs is disordered.The RT-q PCR test results showed that regardless of whether the vascular tissue was ruptured,the expression of AURKA mRNA in ADA tissue was significantly higher than that in the control group.Western blot results showed that the protein level of AURKA was significantly up-regulated in ADA tissues,and the expression was higher in RADA tissues.The results of immunohistochemistry are consistent with the above.3 Compared with the control,the expression of AURKA mRNA in the ADA group increased,and the difference was statistically significant(P<0.05).Compared with the ADA group,the expression of mRNA in the RADA group also increased,and the difference was statistically significant(P<0.05).4 There was no difference between age,sex,history of hypertension,smoking history and drinking history and AURKA expression(P>0.05).And body mass index(X~2=10.700),triglycerides(H=7.773),D-dimer(X~2=3.803),white blood cell count(X~2=10.882),neutrophil count(X~2=15.668)and AURKA expression difference was statistically significant(P<0.05).5 AURKA overexpression vector was transfected in VSMC,and it was found that AURKA mRNA and protein expression levels were significantly up-regulated.Using MTT to analyze cell proliferation,it was found that compared with the GFP control group,overexpression of AURKA significantly promoted the proliferation of VSMC.The results of the scratch healing experiment showed that AURKA overexpression promoted the migration of VSMC.6 Compared with the transfection control si RNA,si-AURKA significantly inhibited the proliferation of VSMC.Scratch healing experiments also show that knocking down AURKA can inhibit cell migration.7 After 24 hours of treatment of VSMC with Ang II,RT-q PCR was used to detect the expression level of AURKA.The results showed that Ang II significantly increased the mRNA expression of AURKA.Western blot results showed that si-AURKA significantly reduced the protein level of AURKA,while Ang II treatment reversed the expression of AURKA inhibited by si-AURKA.MTT and scratch healing experiments showed that the absence of AURKA inhibited the proliferation and migration of VSMC induced by Ang II.8 Western blot results showed that compared with the control si RNA group,there was no change in the expression levels of AKT,p-AKT,ERK,p-ERK and GSK-3βafter AURKA knockout.The protein expression levels of p-GSK-3βand its downstream target geneβ-catenin were significantly down-regulated.RT-q PCR results also showed that AURKA knockdown can reduce the expression ofβ-catenin and PCNA.In order to verify whether Ang II is involved in the GSK-3β/β-catenin pathway regulated by AURKA,this study knocked out and treated VSMC with Ang II,and then detected the expression of GSK-3β,p-GSK-3βandβ-catenin by Western blotting.The results showed that si-AURKA transfection down-regulated the expression levels of p-GSK-3βandβ-catenin,while treatment with Ang II at the same time partially reversed the expression of p-GSK-3βandβ-catenin.These data indicate that the GSK-3β/β-catenin signaling pathway is involved in the proliferation and migration of VSMCs regulated by AURKA.Conclusions 1 AURKA expression is up-regulated in ADA tissues.The up-regulated AURKA promotes and the down-regulated AURKA inhibits the proliferation and migration of VSMCs;2 The expression level of AURKA in ADA tissue is closely related to the incidence of ADA;3 AURKA mediates Ang II to promote VSMC proliferation and migration;4 GSK-3β/β-catenin signaling pathway participates in AURKA-regulated VSMC proliferation and migration.Figure29;Table9;Reference 189.