Study of method assessment of p53 and Bcl-2 gene mRNA expression in rabbit myocardial cells by reverse transcriptase-polymerase chain reaction using cDNA as an internal standard

Objective:To investigate the method of assessment of p53 and Bcl2 gene mRNA in myocardial apoptosis in infarct area in an acute myocardial infarction(AMI) rabbit model.Methods:p53 and Bcl2 gene mRNA expression were assessed by reverse transcriptasepolymerase chain reaction(RTPCR) using cDNA as an internal standard.Results:In the AMI rabbit model,the levels of mRNA expression of p53 gene 〔mRNA/total RNA(ng/g)〕 as assessed by this method were:4hour group(57 648 78±19 776 96) ng/g 6hour group (339 06±104 11) ng/g3hour group(165 44±33 36) ng/g2hour group(88 25± 16 65) ng/g 0hour group(3 16±0 69) ng/g,1 hours group(16 37±2 73) ng/g,8 hours group (23 13±7 03) ng/g ,12 hours group(6 75±2 86) ng/g after coronary artery occlusion (All P 0 05).The levels of expression of Bcl2 mRNA 〔mRNA/total RNA(ng/g)〕 by the method were:3hour group (4 53±1 59) ng/g ,4hour group (0 02±0 01) ng/g and 6hour group (3 46±0 39) ng/g2hour group (52 48±14 18) ng/g ,8hour group (59 24±2 91) ng/g1hour group (77 20±12 48) ng/g and 0hour group (81 77±9 62) ng/g12hour group (99 60±4 71) ng/g after coronary artery occlusion(All P 0 05). This method showed good specificity because the method could assess different quantities of p53 and Bcl2 gene mRNA in the different samples of myocardial cells.The method,moreover,showed high sensitivity because mRNA quantities lower than 0 02 ng/g could be measured.Conclusions:The quantitative method of p53 and Bcl2 gene mRNA expression assessed by RTPCR using cDNA as an internal standard was specific, sensitive, cheap,and simple.It could be performed in an ordinary molecular biology laboratory,so it was worth of recommenting.