title: Decreased mRNA levels of cardiac Cx43 and ZO1 in sudden cardiac death related to coronary atherosclerosis: a pilot study.
abstract: Sudden cardiac death (SCD) is the most frequent cause of sudden unexplained death in forensic practice. The most common cause of SCD is coronary artery disease related to coronary atherosclerosis. Previous study suggested the possible application of connexin 43 (Cx43) and zonula occludens-1 (ZO1) immunostaining in the early diagnosis of myocardial ischemia. However, there appears to be insufficient data with regard to their mRNA levels. The present study investigated the cardiac mRNA levels of Cx43 and ZO1, using forensic autopsy materials consisting of 41 control cases without any disease or structural abnormality of the heart (group 1), 32 deaths due to acute ischemic heart disease related to coronary atherosclerosis without apparent myocardial necrosis (group 2), and 29 traumatic deaths with coronary atherosclerosis (group 3). Ten candidate reference genes were evaluated in the left ventricles of 10 forensic autopsy cases. EEF1A1, PPIA, TPT1, and RPL13A were identified as the most stable reference genes. Using these validated reference genes, mRNA levels of Cx43 and ZO1 were examined in the bilateral ventricles and atria of the heart. Relative mRNA quantification demonstrated decreased calibrated normalized relative quantity (CNRQ) values of Cx43 and ZO1 in bilateral ventricles of group 2. When using one conventional reference gene (GAPDH or ACTB) for normalization, nearly no difference was detected among the three groups. These findings indicate that ventricular gap junction remodeling may be a key contributor to rhythm disturbances. Analysis of cardiac Cx43 and ZO1 using real-time PCR is useful in diagnosis of SCD, and validation of reference genes is crucial.
authors.affiliation: Department of Forensic Medicine, School of Basic Medical Science, Southern Medical University, No. 1838, Guangzhou, 510515, Guangdong Province, China.
authors.forename: Ye
authors.initials: Y
authors.lastname: Xue
authors.affiliation: Department of Forensic Pathology, China Medical University School of Forensic Medicine, Shenyang, China.
authors.forename: Rui
authors.initials: R
authors.lastname: Zhao
authors.affiliation: Department of Forensic Medicine, School of Basic Medical Science, Southern Medical University, No. 1838, Guangzhou, 510515, Guangdong Province, China.
authors.forename: Si-Hao
authors.initials: SH
authors.lastname: Du
authors.affiliation: Collaborative Innovation Center of Judicial Civilization, Beijing, China.|Ministry of Education, Key Laboratory of Evidence Science (China University of Political Science and Law), Beijing, China.
authors.forename: Dong
authors.initials: D
authors.lastname: Zhao
authors.affiliation: Department of Forensic Medicine, School of Basic Medical Science, Southern Medical University, No. 1838, Guangzhou, 510515, Guangdong Province, China.
authors.forename: Dong-Ri
authors.initials: DR
authors.lastname: Li
authors.affiliation: Department of Forensic Medicine, School of Basic Medical Science, Southern Medical University, No. 1838, Guangzhou, 510515, Guangdong Province, China.
authors.forename: Jing-Tao
authors.initials: JT
authors.lastname: Xu
authors.affiliation: Department of Toxicology, School of Public Health and Tropical Medicine, Southern Medical University, No. 1838, Guangzhou, 510515, Guangdong Province, China. xiexiaoli1999@126.com.
authors.forename: Xiao-Li
authors.initials: XL
authors.lastname: Xie
authors.affiliation: Department of Forensic Medicine, School of Basic Medical Science, Southern Medical University, No. 1838, Guangzhou, 510515, Guangdong Province, China. wangqi_legmed@126.com.
authors.forename: Qi
authors.initials: Q
authors.lastname: Wang
mesh_terms: D016022:Case-Control Studies; D018031:Connexin 43 / Q000235:genetics*; D003324:Coronary Artery Disease / Q000235:genetics* / Q000378:metabolism; D016757:Death, Sudden, Cardiac / Q000473:pathology*; D005260:Female; D053803:Forensic Genetics; D006325:Heart Atria / Q000378:metabolism; D006352:Heart Ventricles / Q000378:metabolism; D006801:Humans; D008297:Male; D009206:Myocardium / Q000378:metabolism; D010865:Pilot Projects; D012333:RNA, Messenger / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D000090324:Tumor Protein, Translationally-Controlled 1; D062826:Zonula Occludens-1 Protein / Q000235:genetics*
chemical_list: D018031:Connexin 43; D012333:RNA, Messenger; C000715647:TPT1 protein, human; D000090324:Tumor Protein, Translationally-Controlled 1; D062826:Zonula Occludens-1 Protein
keywords: Cx43; Forensic molecular pathology; Reference gene selection; Sudden cardiac death; Ventricular arrhythmia; ZO1