[Construction of the recombinant adenovirus vector carrying antisense multidrug resistance-associated protein].
OBJECTIVE
The recombinant adenovirus vector carrying antisense multidrug resistance-associated protein (MRP) was constructed for use in gene therapy to overcome drug resistance in hepatocellular carcinoma.


METHODS
The fragment of MRP gene encoding 5' region was cloned reversely into the shuttle plasmid pAdTrack-CMV, with the resultant plasmid and the backbone plasmid pAdEasy-1, the homologous recombination took place in the E. coli BJ5183 and the recombinant adenoviral plasmid was generated. The adenoviruses were packaged and amplified in the 293 cells. Then the viral titer was checked by GFP.


RESULTS
The recombinant adenovirus vector carrying antisense MRP was constructed successfully. The viral titer was 2.5 x 10(9).


CONCLUSIONS
The recombinant adenovirus vector constructed by us could introduce the antisense MRP into the human drug-resistant hepatocellular cell line effectively, which would provide experimental basis for the mechanisms and reversal methods of the multidrug resistance in human hepatocellular carcinoma.
Chen
Lin
L
Department of General Surgery, West China Hospital, Sichuan University, Chengdu 610041, China.
Gou
Xinghua
X
Yan
Lülan
L
Han
Lei
L
Li
Dehua
D
Zhao
Yongheng
Y
Hu
Haiyang
H
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000096:biosynthesis / Q000235:genetics; D000256:Adenoviridae / Q000235:genetics* / Q000378:metabolism; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000628:therapy*; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005333:Fetus; D018014:Gene Transfer Techniques; D019450:Genes, MDR / Q000235:genetics; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D007668:Kidney / Q000166:cytology; D008113:Liver Neoplasms / Q000235:genetics / Q000628:therapy*; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000096:biosynthesis / Q000235:genetics*; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D011995:Recombination, Genetic
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D016376:Oligonucleotides, Antisense; D011994:Recombinant Proteins