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Specific inhibition of gene expression of lung resistance-related protein by short interfering RNA / 南方医科大学学报
Journal of Southern Medical University ; (12): 1-5, 2006.
Article in English | WPRIM | ID: wpr-234208
ABSTRACT
<p><b>OBJECTIVE</b>To investigate inhibitory effect of short interfering RNA (siRNA) on the expression of lung resistance-related protein (LRP) in leukemia cells.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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 level defined as negative results; the GFP expression showed no significant difference between the cells transfected 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 transfected with pcDNA3.0/LRP and those co-transfected with GFP gene-specific siRNA and pcDNA3.0/LRP.</p><p><b>CONCLUSIONS</b>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.</p>
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Full text: Available Index: WPRIM (Western Pacific) Main subject: RNA, Messenger / Transfection / Genetic Therapy / Drug Resistance, Multiple / K562 Cells / Vault Ribonucleoprotein Particles / RNA, Small Interfering / Green Fluorescent Proteins / Genetics Type of study: Practice guideline Limits: Humans Language: English Journal: Journal of Southern Medical University Year: 2006 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: RNA, Messenger / Transfection / Genetic Therapy / Drug Resistance, Multiple / K562 Cells / Vault Ribonucleoprotein Particles / RNA, Small Interfering / Green Fluorescent Proteins / Genetics Type of study: Practice guideline Limits: Humans Language: English Journal: Journal of Southern Medical University Year: 2006 Type: Article