PMID- 27455553
OWN - NLM
STAT- MEDLINE
DCOM- 20160817
LR  - 20181202
IS  - 0031-7144 (Print)
IS  - 0031-7144 (Linking)
VI  - 71
IP  - 6
DP  - 2016 Jun
TI  - Construction of HEK293 cells stably expressing wild-type organic anion 
      transporting polypeptide 1B1 (OATP1B1*1a) and variant OATP1B1*1b and OATP1B1*15.
PG  - 337-9
AB  - A transgenic cell line stably expressing the human organic anion transporting 
      polypeptide (OATP1B1) was established. Human Embryonic Kidney 293 (HEK293) cell 
      line stably expressing OATP1B1*1a sequence was amplified through PCR with the 
      extracted total RNA as templates from human liver, then subcloned into the 
      plasmid pMD19-T and verified by sequencing. OATP1B1*1b/OATP1B1*15 mutant 
      sequences were obtained by site-directed mutation PCR with pMD19-T/ OATP1B1*1a as 
      templates. The plasmids pcDNA3.1(+)/OATP1B1*1a, *1b and *15 were constructed and 
      transfected into HEK293 cell line using Lipofectamine 2000 transfection reagent. 
      Several stable transfected clones were obtained after selection with G418. Using 
      rosuvastatin as a probe substrate of OATP1B1, the intracellular rosuvastatin 
      accumulation in HEK293 and HEK-OATP1B1*1a, *1b and *15 monoclone cells were 
      validated by a ultra-performance liquid chromatography-tandem mass spectrometry. 
      OATP1B1 mRNA and protein expression were detected by RT-PCR and Western blot, 
      respectively. The results from RT-PCR, rosuvastatin uptake and Western blot assay 
      indicated that human OATP1B1 was highly expressed in transfected cells compared 
      with controls. The HEK-293 cell lines stably expressing human OATP1B1-wild and 
      variant (HEK-OATP1B1, *1b and *15) are potential models to study drug transport 
      in vitro.
FAU - Chen, M
AU  - Chen M
FAU - Qu, B X
AU  - Qu BX
FAU - Chen, X L
AU  - Chen XL
FAU - Hu, H H
AU  - Hu HH
FAU - Jiang, H D
AU  - Jiang HD
FAU - Yu, L S
AU  - Yu LS
FAU - Zhou, Q
AU  - Zhou Q
FAU - Zeng, S
AU  - Zeng S
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Germany
TA  - Pharmazie
JT  - Die Pharmazie
JID - 9800766
RN  - 0 (Hydroxymethylglutaryl-CoA Reductase Inhibitors)
RN  - 0 (Lipids)
RN  - 0 (Lipofectamine)
RN  - 0 (Liver-Specific Organic Anion Transporter 1)
RN  - 0 (Organic Anion Transporters)
RN  - 0 (RNA, Messenger)
RN  - 0 (SLCO1B1 protein, human)
RN  - 83MVU38M7Q (Rosuvastatin Calcium)
SB  - IM
MH  - Cell Membrane/chemistry/metabolism
MH  - Genetic Variation
MH  - HEK293 Cells/*metabolism
MH  - Humans
MH  - Hydroxymethylglutaryl-CoA Reductase Inhibitors/metabolism
MH  - Lipids
MH  - Liver/metabolism
MH  - Liver-Specific Organic Anion Transporter 1
MH  - Mutagenesis, Site-Directed
MH  - Organic Anion Transporters/*biosynthesis/*genetics
MH  - Plasmids/genetics
MH  - RNA, Messenger/biosynthesis/genetics
MH  - Rosuvastatin Calcium/metabolism
MH  - Transfection/*methods
EDAT- 2016/07/28 06:00
MHDA- 2016/08/18 06:00
CRDT- 2016/07/27 06:00
PHST- 2016/07/27 06:00 [entrez]
PHST- 2016/07/28 06:00 [pubmed]
PHST- 2016/08/18 06:00 [medline]
PST - ppublish
SO  - Pharmazie. 2016 Jun;71(6):337-9.