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Published on: February 9, 2024
[Construction of the recombinant adenovirus vector carrying antisense multidrug resistance-associated protein]
Lin Chen1, Xinghua Gou, Lülan Yan
1Department of General Surgery, West China Hospital, Sichuan University, Chengdu 610041, China.
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).
Conclusion:
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.
