Study on inhibition effect of small interfering RNA on monocyte chemotacite protein-1 in HKC / 中国实用内科杂志

ABSTRACT

Objective To investigate the inhibition effect of plasma-mediated small interfering RNA (siRNA) on the expression of monocyte chemotacite protein-1 (MCP-1) gene in human renal tubular epithelial cells (HKC).Methods Three pairs of siRNAs directed human's MCP-1 mRNA 67,116,142 targets were designed and synthesized.Eukaryotic expression vector special for MCP-1,pRNAT-MCP-1-Ⅰ、pRNAT-MCP-1-Ⅱ、pRNAT-MCP-1-Ⅲ were constructed and transfected into HKC by lipofectamine.At 24 hour after transfection,the expression of MCP-1 in the levels of mRNA was detected by Real Time RT-PCR,and the expression of MCP-1 in the levels of protein was detected by Western blot.Results Transfection efficiency of siRNA expression vector was 60%;the expression of MCP-1 in the levels of mRNA and protein of three pairs of plasma-mediated siRNA group were markedly decreased compared with normal control group(P