ABCMdb

PMID: 18710444

Gardner ER, Ahlers CM, Shukla S, Sissung TM, Ockers SB, Price DK, Hamada A, Robey RW, Steinberg SM, Ambudkar SV, Dahut WL, Figg WD
Association of the ABCG2 C421A polymorphism with prostate cancer risk and survival.
BJU Int. 2008 Dec;102(11):1694-9. Epub 2008 Aug 14., [PubMed]

Sentences

No. Mutations Sentence Comment

5 ABCG2 p.Gln141Lys Intracellular accumulation of PhIP was 80% higher in HEK293 cells transfected with Q141K ABCG2 than in wild-type cells, confirming that this SNP decreases transport of PhIP. Login to comment 10 ABCG2 p.Gln141Lys A SNP of ABCG2, C421A, resulting in a glutamine to lysine change at amino acid 141, has been shown to result in decreased function of the protein. Login to comment 43 ABCG2 p.Gln141Lys ABCG2 p.Gln141Lys CELL LINES Human embryonic kidney cells (HEK293) stably transfected with empty pcDNA3.1 vector (pcDNA3.1-HEK) or pcDNA3.1 vector containing full-length wild-type (482R-HEK) or Q141K (Q141K-HEK) ABCG2 have been previously characterized and were maintained as described [18,27]. Login to comment 46 ABCG2 p.Gln141Lys [14 C]-PhIP AND [3 H]-TESTOSTERONE ACCUMULATION ASSAYS pcDNA3.1-HEK, R482-HEK and Q141K-HEK cells (2.5 × 105 cells/well) were grown in monolayer in a 24-well tissue culture plate at 37 °C in the presence of 5% CO2 in Dulbecco`s Modified Eagle`s Medium supplemented with 10% FBS. Login to comment 49 ABCG2 p.Gln141Lys DETERMINATION OF CELL SURFACE EXPRESSION OF ABCG2 The cell surface expression of ABCG2 in pcDNA3.1-HEK, R482-HEK and Q141K-HEK cells was determined as described previously [29]. Login to comment 69 ABCG2 p.Gln141Lys EFFECT OF C421A SNP ON IN VITRO PhIP TRANSPORT Comparison of the pcDNA3.1-HEK cells that do not express ABCG2 with the R482-HEK cells that express wild-type ABCG2 showed a three-fold decrease in intracellular accumulation of [14 C]-PhIP, in the absence of FTC (P < 0.001) as shown in Fig. 2a. The cells expressing the C421A variant form of ABCG2 (Q141K-HEK) had a reduced ability to efflux [14 C]-PHIP, resulting in a 1.8-fold increase in intracellular accumulation of this substrate compared with the wild-type cells (P < 0.001). Login to comment 71 ABCG2 p.Gln141Lys Comparative ABCG2 expression levels were confirmed in the R482-HEK and Q141K-HEK cell lines, using flow cytometry as shown in Fig. 2b. Login to comment 72 ABCG2 p.Gln141Lys EFFECT OF C421A SNP ON IN VITRO TESTOSTERONE TRANSPORT Transport assays using transfected R482-HEK and Q141K-HEK cells showed that testosterone was not transported by either TABLE 1 ComparisonofgenotypicfrequenciesofABCG2C421ASNPinpatientsdiagnosedwithAIPCand healthy volunteers (control). Login to comment 89 ABCG2 p.Gln141Lys ABCG2 p.Gln141Lys Green and red bars represent testosterone accumulation with and without the presence of FTC, respectively. pcDNA3.1-HEK cells do not express ABCG2. R482-HEK cells express wild-type ABCG2 and Q141K-HEK cells are homozygous variant for the C421A SNP. Each bar represents the mean accumulation (n = 3) and the error bars indicate the SD. 50 40 30 20 10 0 pcDNA3.1-HEK R482-HEK Q141K-HEK FIG. 4. Login to comment 94 ABCG2 p.Gln141Lys C421ASNPresultsindecreasedintracellularPhIPaccumulation in vitro.Greenandredebarsrepresent PhIP accumulation with and without the presence of FTC, respectively. pcDNA3.1-HEK cells do not express ABCG2. R482-HEK cells express wild-type ABCG2 and Q141K-HEK cells are homozygous variant for the C421A SNP. Each bar represents the mean accumulation (n = 3) and the error bars indicate the SD. Login to comment 96 ABCG2 p.Gln141Lys ABCG2 p.Gln141Lys (a) (b) 1400 1200 1000 800 600 400 200 0 pcDNA3.1-HEK R482-HEK Q141K-HEK 14C-Phipaccumulation, nmoles/millioncells pcDNA3.1-HEK R482-HEK FL2-Height 0 30 60 90 120 150 0 30 60 90 120 150 0 30 60 90 120 150 2000 400 600800 1000 2000 400 600 800 1000 2000 400 600 800 1000 CountsCountsCounts FL2-Height FL2-Height Q141K-HEK accumulation of docetaxel, leading to improved response. Login to comment