Reversal of mdr1b-dependent multidrug resistance in a rat astrocyte model by adenoviral-delivered short hairpin RNA.
Over-expression of P-glycoprotein (Pgp), a protein responsible for multidrug resistance (MDR), is responsible for general resistance to anti-epileptic drugs (AEDs). We explored the potential use of gene therapy with adenoviral-delivered RNA interference against mdr1b as a method to sensitize refractory epilepsy to AEDs. We constructed replication-deficient recombinant adenovirus Adeno-mdr1b1 carrying short hairpin RNA (shRNA) targeting against mdr1b, and successfully infected the established Sprague-Dawley rat astrocyte model of Coriaria Lactone-induced Pgp over-expression. The expression levels of mdr1b and Pgp and the Rhodamine123 efflux ratio in trial groups were significantly lower than that of blank control (P < 0.05) during the first 7 days post-infection, with the most inhibition at 48 h. The results suggest that knockdown of MDR using adenovirus not only avoided the toxicity and low rate of plasmid nucleofection, but also overcame its poor efficiency of mdr1b silencing. More importantly, this study may pave the way for a promising approach to remedy refractory epilepsy.
Chen
Lei
L
Department of Neurology, West China Hospital, Sichuan University, Chengdu, Sichuan, People's Republic of China. leilei_25@126.com
Tian
Linyu
L
Yang
Tianhua
T
Cheng
Xinwang
X
Hermann
Stefan
S
Zhou
Dong
D
D018435:ATP Binding Cassette Transporter, Subfamily B / Q000235:genetics*; D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000166:cytology / Q000378:metabolism*; D002460:Cell Line; D018432:Drug Resistance, Multiple / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D008156:Luciferases / Q000378:metabolism; D008954:Models, Biological; D010957:Plasmids / Q000235:genetics; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D020112:Rhodamine 123 / Q000378:metabolism; D000097202:ATP-Binding Cassette Sub-Family B Member 4
D018435:ATP Binding Cassette Transporter, Subfamily B; D049452:Green Fluorescent Proteins; D008156:Luciferases; D034741:RNA, Small Interfering; D020112:Rhodamine 123; D000097202:ATP-Binding Cassette Sub-Family B Member 4