ObjectiveTo construct the recombinant adenovirus vector carrying antisense multidrug resistanceassociated protein (MRP) and transfect the human drugresistant hepatocellular carcinoma cell line(SMMC7721/ADM). MethodsThe fragment of MRP gene encoding 5′region was cloned reversely into the shuttle plasmid pAdTrackCMV, with the resultant plasmid and the backbone plasmid pAdEasy1,the homologous recombination took place in the bacteria and the recombinant adenoviral plasmid was generated. The adenoviruses were packaged and amplified in 293 cells. Then the cell line of SMMC7721/ADM was transfected with the resultant adenoviruses.ResultsThe recombinant adenovirus vector carrying antisense MRP was constructed successfully. The viral titer was 2.5×109 efu/ml, and more than 90% SMMC7721/ADM cells could be transfected when the multiplicity of infection（MOI） was 100. ConclusionThe recombinant adenovirus vector constructed by us could introduce the antisense MRP into the human drugresistant hepatocellular cell line effectively, which would provide experimental basis for the mechanisms and reversal methods of the multidrug resistance in human hepatocellular carcinoma.
Citation：
CHEN Lin,GOU Xinghua,YAN Lnan,ZHAO Yongheng,HAN Lei,LI Dehua,HU Haiyang,ZHAO Lanying. Construction of the Recombinant Adenovirus Carrying Antisense Multidrug ResistanceAssociated Protein and the Study of Its Application. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2003, 10(2): 117-120. doi:
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- 1. Cole SP, Bhardwaj G, Gerlach JH, et al. Overexpression of a transporter gene in a multidrugresistant human lung cancer cell line [Ｊ]. Science， 1992; 258(5088)∶1650.
- 2. Hipfner DR,Deeley RG,Cole SP.Structural, mechanistic and clinical aspects of MRP1 [Ｊ]. Biochim Biophys Acta,1999; 1461(2)∶359.
- 3. Roelofsen H, Vos TA, Schippers IJ, et al. Increased levels of the multidrug resistance protein in lateral membranes of proliferating hepatocytederived cells [Ｊ]. Gastroenterology, 1997; 112(2)∶ 511.
- 4. 戴越盟，林琦远，严律南,等. 多药耐药相关蛋白在阿霉素诱导人肝癌细胞SMMC7721耐药性产生中的作用及机理 [Ｊ]. 中国普外基础与临床杂志，2001； 8（1）∶8.
- 5. Carter G, Lemoine NR. Antisense technology for cancer therapy: does it make sense? [Ｊ]. Br J Cancer, 1993; 67(5)∶869.
- 6. He TC,Zhou S,da Costa LT,et al.A simplified system for generating recombinant adenoviruses [Ｊ].Proc Natl Acad Sci USA,1998; 95(5)∶2509.
- 7. Inoue H, Nojima H, Okayama H. High efficiency transformation of Escherichia coli with plasmids [Ｊ]. Gene, 1990; 96(1)∶23.
- 8. 司徒振强，吴军正主编. 细胞培养 [M]. 第1版.西安： 世界图书出版公司，1996∶186～187.
- 9. Itsubo M,Ishikawa T,Toda G,et al.Immunohistochemic study of expression and cellular localization of the multidrug resistance gene product Pglycoprotein in primary liver carcinoma [Ｊ].Cancer,1994; 73(2)∶ 298.
- 10. Lai EC, Choi TK, Cheng CH, et al. Doxorubicin for unresectable hepatocellular carcinoma. A prospective study on the addition of verapami [Ｊ]. Cancer, 1990; 66(8)∶1685.
- 11. Bradshaw DM, Arceci RJ. Clinical relevance of transmembrane drug efflux as a mechanism of multidrug resistance [Ｊ]. J Clin Oncol, 1998; 16(11)∶3674.
