The evaluation of efflux transporter model based on RNA interference technology in vitro / 药学学报
Acta Pharmaceutica Sinica
;
(12): 1122-1127, 2015.
Article
in Chinese
| WPRIM
| ID: wpr-257018
ABSTRACT
In the present study, the specifically knockdown models of P-gp or MRP2 were constructed by using a series of chemically synthesized small interfering RNA (siRNA) in vitro. The expression of P-gp and MRP2 was measured by real-time PCR and Western blot, and the function was evaluated by applying P-gp and MRP2 substrate, rhodamine and methotrexate. The results showed that MRP2 siRNA-3 or P-gp siRNA-2 significantly decreased the mRNA expression of MRP2 or P-gp, the inhibition ratio was 68% or 84%; MRP2 siRNA-3 or P-gp siRNA-2 at a dose of 80 nmol x L(-1) significantly reduced the protein expression of MRP2 or P-gp at 48 h after treatment, the inhibition ratio was 62% or 70%. Meanwhile, other transporters were not influenced by siRNA. When pretreatment with MRP2 siRNA-3 or P-gp siRNA-2, the efflux of methotrexate or rhodamine decreased significantly and the intra-cellular concentration increased. The results suggested that chemically synthesized siRNA could significantly inhibit the expression and function of MRP2 and P-gp, and the model of RNAi in vitro could be used to evaluate the role of efflux transporters in transportation of drugs.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
ATP Binding Cassette Transporter, Subfamily B, Member 1
/
Multidrug Resistance-Associated Proteins
/
RNA, Small Interfering
/
RNA Interference
/
Gene Knockdown Techniques
/
Real-Time Polymerase Chain Reaction
/
Genetics
Language:
Chinese
Journal:
Acta Pharmaceutica Sinica
Year:
2015
Type:
Article
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