title: [Establishment of BCRP expressed pig kidney cell line LLC-PK1/BCRP and its biological profile].
abstract: To establish a pig kidney cell line LLC-PK1/BCRP in which human breast cancer resistance protein was highly expressed, the expression vector pcDNA3.1(+)-BCRP which contained BCRP gene was constructed and transfected into LLC-PKI cells via liposomes. After selecting with G418, population doubling time, flow cytometry and Western blotting analysis were used to evaluate the cell line. MTT assays were employed to determine the drug resistance index of mitoxantrone and doxorubicin. Invert fluorescent microscope was used to observe the efflux of fluorescence dye Hoechst 33342 by BCRP, furthermore, the BCRP's inhibitor GF120918 was applied to reverse the efflux of Hoechst 33342. The experiment results showed that the expression of BCRP protein increased in LLC-PK1/BCRP cell. The population doubling time of LLC-PK1/BCRP cell was a little longer than that of the parental cell LLC-PK1. The resistance indexes to mitoxantrone and doxorubicin were 51.95 and 6.09 times, respectively, higher than LLC-PK1 cell. The efflux of Hoechst 33342 was significantly enhanced and could be reversed by GF120918. So a LLC-PK1/BCRP cell line was established, which highly expressed BCRP protein successfully. This cell line could be a valuable model to further investigate the biological profile of BCRP and select the substrate and inhibitor of BCRP.
authors: Department of Pharmaceutical Analysis and Drug Metabolism, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China.
authors: Ye
authors: Y
authors: Tian
authors: Bo-Xuan
authors: BX
authors: Qu
authors: Yan
authors: Y
authors: Yao
authors: Su
authors: S
authors: Zeng
mesh_terms: D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters / Q000235:genetics / Q000378:metabolism*; D000166:Acridines / Q000494:pharmacology; D000818:Animals; D001562:Benzimidazoles / Q000378:metabolism; D002453:Cell Cycle; D049109:Cell Proliferation; D004317:Doxorubicin / Q000494:pharmacology; D018432:Drug Resistance, Multiple; D005822:Genetic Vectors; D018374:LLC-PK1 Cells / Q000166:cytology / Q000378:metabolism; D008942:Mitoxantrone / Q000494:pharmacology; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids; D013552:Swine; D044005:Tetrahydroisoquinolines / Q000494:pharmacology; D014162:Transfection
chemical_list: C116614:ABCG2 protein, human; D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters; D000166:Acridines; D001562:Benzimidazoles; D009363:Neoplasm Proteins; D044005:Tetrahydroisoquinolines; D004317:Doxorubicin; D008942:Mitoxantrone; C083501:Elacridar; C017807:bisbenzimide ethoxide trihydrochloride