SERCA2 and ANF Promoter-Activity Studies in Hypertrophic Cardiomyocytes Using Liposome-, Gene Gun-, and Adenovirus-Mediated Gene Transfer

摘要: Myocardial hypertrophy is known as the process of enlargement ventricular cells, which also accompanied by changes in phenotype. The latter include, e.g., downregulation expression sarcoplasmatic reticulum Ca2+ ATPase (SERCA2), phospholamban (PL), and β-adrenergic receptor upregulation atrial natriuretic factor (ANF) β-myosin heavy chain (s-MHC). Analysis transcriptional regulation a promoter fragment SERCA2 gene using liposome-mediated transfection revealed that may not respond to general increase transcription upon stimulation neonatal rat cardiomyocytes 10-8M endothelin-1. Liposome-mediated used these activity studies yields no more than 1% efficiency (the percentage cells expressing transgene). To obtain higher efficiency, we set out develop gene-gun biolistics method for cardiomyocytes. An up approximately 10% can be achieved gun tested RSV-β-Gal construct. Here, demonstrate efficacy use an endothelin-1 inducible ANF coupled CAT reporter. Therefore, ideally suited quick reliable test DNA constructs on their activity. normally only active very low extent adult cells; it upregulated hypertrophic stimuli. We property generating encompassing antisense PL under control fragment. Adenovirus infection with almost 100% required measure functional consequences overexpressed PL. present first results ANF-promoter-PL-antisense adenovirus