Real-time quantitative RT-PCR in measuring Siglec-1 gene expression levels in peripheral blood mononuclear cells of patients with coronary heart disease / 第二军医大学学报

ABSTRACT

Objective: To establish a real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) method for examining the expression of sialic acid-binding immunoglobulin-like lectin-one (Siglec-1)gene in the peripheral blood mononuclear cells (PBMCs) in patients with coronary heart disease (CHD),so as to explore the relationship between Siglec-1 mRNA expression and the development/progression of CHD. Methods: Based on the fluorescent SYBR Green method, a real-time quantitative RT-PCR was set up. The expression of Siglec-1 gene in the PBMCs of 57 patients with CHD and 38 healthy controls were measured by ABI PRISM 7000 Sequence Detection Systems. The blood lipid levels were determined in all the subjects by routine biochemical examination. Results: The mean Siglec-1 mRNA copy was significantly higher in the CHD group than that in the healthy control group (mean 3.23 folds, ranging 1.28-8.11 folds, P < 0.01). The expression in the acute myocardial infarction group,stable angina and unstable angina group were 3.32(1.38-7.97),2.56(0.88-7.42),and 3.35(1.25-8.96) folds that of the control group,respectively. No significant difference was observed in the normalized Siglec-1 mRNA copy number between CHD group with normal level of serum lipids and abnormal level of serum lipids. Conclusion: A real-time quantitative RT-PCR method for detecting the expression of Siglec-1 gene in PBMCs has been successfully established. The expression of Siglec-1 is dramatically increased in PBMCs in CHD patients.