Specific inhibition of gene expression of lung resistance-related protein by short interfering RNA
10.3321/j.issn:1673-4254.2006.01.001
- VernacularTitle:短干涉RNA特异性抑制白血病细胞中肺耐药相关蛋白基因表达
- Author:
Ning LI
1
;
Xin-Hua QIAN
;
Zhi-Yuan WANG
Author Information
1. 南方医科大学附属南方医院
- Keywords:
lung resistance-related protein;
leukemia;
gene therapy,RNA interference
- From:
Journal of Southern Medical University
2006;26(1):1-5
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate inhibitory effect of short interfering RNA (siRNA) on the expression of lung resistance-related protein (LRP) in leukemia cells. Methods The eukaryotic vectors of LRP, pcDNA3.0/LRP, were constructed. The transfection protocol of K562 cells grown in standard conditions consisted of different combinations of pcDNA3.0/LRP, pEGFP-C1 expressing mammalian enhanced green fluorescent protein (GFP), and their gene-specific siRNAs. RT-PCR and flow cytometry were employed to evaluate the mRNA and protein expression of LRP and fluoroscopy was performed for assay of GFP expression in the transfected cells. Results Compared with untreated K562 cells, pcDNA3.0/LRP-transfected cells showed increased LRP mRNA and protein expression and the positive cell percentage reached 30%. In the cells co-transfected with LRP gene-specific siRNA and pcDNA3.0/LRP, both LRP mRNA and protein expression decreased significantly to a lev el defined as negative results; the GFP expression showed no significant difference between the cells transfcted with pEGFP-C1 and those co-transfected with LRP gene-specific siRNA and pEGFP-C1. LRP mRNA and protein expressions were also similar between the cells transfcted with pcDNA3.0/LRP and those co-transfected with GFP gene-specific siRNA and pcDNA3.0/LRP. Conclusions The LRP gene-specific siRNA we designed is capable of degrading LRP mRNA and inhibiting the protein expression effectively and specifically, which shed light on the potential application of siRNA for gene-specific therapy to reverse LRP-induced multidrug resistance of leukemia cells.
