ABCB1 p.Pro32Asp
| Predicted by SNAP2: | A: N (66%), C: N (53%), D: N (57%), E: N (66%), F: N (57%), G: N (61%), H: N (72%), I: N (66%), K: N (82%), L: N (61%), M: N (61%), N: N (78%), Q: N (82%), R: N (61%), S: N (78%), T: N (72%), V: N (61%), W: D (63%), Y: N (57%), |
| Predicted by PROVEAN: | A: N, C: D, D: N, E: N, F: D, G: D, H: N, I: D, K: N, L: D, M: N, N: N, Q: N, R: N, S: N, T: N, V: N, W: D, Y: D, |
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[hide] Effect of cell differentiation and passage number ... Eur J Pharm Biopharm. 2007 Sep;67(2):548-54. Epub 2007 Mar 28. Siissalo S, Laitinen L, Koljonen M, Vellonen KS, Kortejarvi H, Urtti A, Hirvonen J, Kaukonen AM
Effect of cell differentiation and passage number on the expression of efflux proteins in wild type and vinblastine-induced Caco-2 cell lines.
Eur J Pharm Biopharm. 2007 Sep;67(2):548-54. Epub 2007 Mar 28., [PMID:17475454]
Abstract [show]
The mRNA level expression of MDR1, MRP1-6, BCRP and CYP3A4 was determined by quantitative PCR in wild type (Caco-2WT) and vinblastine-treated (Caco-2VBL) Caco-2 cells at different passage levels (32-53). Differentiation increased the mRNA levels of MDR1, BCRP and all the MRPs except MRP4. Corresponding mRNA levels were observed in Caco-2WT and Caco-2VBL, except that the expression of MRD1 was higher in Caco-2VBL than in Caco-2WT cells. CYP3A4 was barely detected in either cell line. MDR1 functionality was studied using rhodamine123 and verapamil as a substrate-inhibitor pair. Corresponding to the observed differences in mRNA levels, MDR1 activity was higher in the Caco-2VBL cells. In Caco-2WT, MDR1 functionality was elevated at low passage numbers (32-35) compared to higher ones (49-53). Verapamil inhibited MDR1 efflux except at higher passage Caco-2WT cells, where no MDR1 activity could be observed. The results support the use of Caco-2VBL cells in MDR1 screening. The functional expression is higher than in Caco-2WT and remains consistent across the studied passages without major differences in mRNA levels of other efflux proteins. As both the passage number and the level of cell differentiation affect the expression profile of efflux proteins, short-term cell growth protocols should be evaluated accordingly.
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No. Sentence Comment
112 0 0.005 0.01 0.015 0.02 0.025 0.03 0.035 Caco-2WT P32D/ P33U Caco-2WT P33D/ P32U Caco-2WT P43D/ P44U Caco-2VBL P33/9D/ P35/11U Caco-2VBL P42/13D/ P43/14U Caco-2VBL P48/19D/ P49/20U Relativeexpression Diff Undiff Fig. 2. mRNA expression level of MDR1 vs. b-actin in differentiated (D, Diff) and undifferentiated (U, Undiff) Caco-2WT and Caco-2VBL samples (n = 3) over the passage range typically used for transport experiments in our laboratory.
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ABCB1 p.Pro32Asp 17475454:112:50
status: NEW