Construction of p300 specific siRNA vector and its effect on GATA4 expression in cardiac muscle cells / 第三军医大学学报

ABSTRACT

Objective To construct the p300 specific siRNA vector in mice and observe its effect on cardiac transcription factor,GATA4,for further study of the effect of p300-mediated histone acetylation on heart growth. Methods Three recombinant plasmid vectors ( p300 RNAi1,p300 RNAi2,and p300 RNAi3s) ,designed and constructed according to the conservative sequence of mice p300,were respectively tansfected into in vitro cultured cardiac muscle cells of suckling mice. Expression of p300 and GATA4 at mRNA and protein levels was detected by RT-PCR and immunofluorescence,respectively. Results The expression level of p300 mRNA was not significantly decreased in pSOS-HUS and p300 RNAi2 groups,but significantly decreased in p300 RNAi1 and p300 RNAi3 groups ( 0. 220 8 ? 0. 020 0 and 0. 170 8 ? 0. 040 0 vs 0. 509 5 ? 0. 030 0,P