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BMC Medical Genetics

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Open Access 01-12-2006 | Research article

The GCC repeat length in the 5'UTR of MRP1 gene is polymorphic: a functional characterization of its relevance for cystic fibrosis

Authors: Elena Nicolis, Matteo Pasetto, Cristina Cigana, Ugo Pradal, Baroukh M Assael, Paola Melotti

Published in: BMC Medical Genetics | Issue 1/2006

Abstract

Background

Among the members of the ATP binding cassette transporter superfamily, MRPs share the closest homology with the CFTR protein, which is defective in CF disease. MRP1 has been proposed as a potential modifier gene and/or as novel target for pharmacotherapy of CF to explain the clinical benefits observed in some CF patients treated with the macrolide AZM. The 5'UTR of the MRP1 gene contains a GCC triplet repeat that could represent a polymorphic site and affect the activity of the promoter.

Methods

The MRP1 5' flanking region was amplified by PCR from 36 CF patients and 100 non-CF subjects and the number of GCC triplets of each allele was determined by sequence and electrophoretic analysis. We performed gene reporter studies in CF airway epithelial cells 16HBE14o-AS3, in basal conditions and in the presence of AZM.

Results

We found that the GCC repeat is polymorphic, ranging from 7 to 14 triplets either in CF or in non-CF subjects. Our data are preliminary and have to be confirmed on a larger population of CF subjects. The transcriptional activity of the proximal MRP1 5' regulatory region revealed no statistically significant correlations between the number of repeats and treatment with AZM.

Conclusion

We identified a novel polymorphism in the 5'UTR of MRP1 gene that provides multiple alleles in a gene relevant for multidrug resistance as well as for CF, determining that this region is transcriptionally active and that this activity does not appear to be influenced by AZM treatment.

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Lindsell PHJ: Glutathione permeability of CFTR. Am J Physiol. 1998, 275: C323-

Gao LKK, Yankaskas JR, Forman HJ: Abnormal glutathione transport in cystic fibrosis airway epithelia. Am J Physiol. 1999, 277: L113-PubMed

Altschuler EL: Azithromycin, the multidrug-resistant protein, and cystic fibrosis. Lancet. 1998, 351: 1286-10.1016/S0140-6736(05)79350-8.CrossRefPubMed

Lallemand JY, Stoven V: Induction by antitumoral drugs of proteins that functionally complement CFTR: a novel therapy for cystic fibrosis?. Lancet. 1997, 350: 711-712. 10.1016/S0140-6736(05)63510-6.CrossRefPubMed

Hurbain I, Sermet-Gaudelus I: Evaluation of MRP1-5 gene expression in cystic fibrosis patients homozygous for the delta F508 mutation. Pediatr Res. 2003, 54: 627-634. 10.1203/01.PDR.0000090926.16166.3F.CrossRefPubMed

Pradal U, Cipolli M, Cazzola G: Chloride transport may be restored by long-term azithromycin treatment in patients with cystic fibrosis. Pediatr Pulmonol. 2000, 280-Suppl 20

Pradal U, Delmarco A: Long term azithromycin in cystic fibrosis: another possible mechanism of action?. J Chemother. 2005, 17: 384-400.CrossRef

Pradal U, Delmarco A: Azithromycin induces over-expression of multidrug resistance-associated protein (MRP) and restores chloride conductance in cystic fibrosis [abstract]. J Cystic Fibrosis. 2002, 1: s52-

Zhu Q, Center MS: Cloning and sequence analysis of the promoter region of the MRP gene of HL60 cells isolated for resistance to adriamycin. Cancer Res. 1994, 54: 4488-4492.PubMed

10.

Kruh GD, Gaughan KT: Expression pattern of MRP in human tissues and adult solid tumor cell lines. J Natl Cancer Inst. 1995, 87: 1256-1258.CrossRefPubMed

11.

Dean M, Rzhetsky A: The human ATP-binding cassette (ABC) transporter superfamily. Genome Res. 2001, 11: 1156-1166. 10.1101/gr.GR-1649R.CrossRefPubMed

12.

Loe DW, Deeley RG: Biology of the multidrug resistance-associated protein, MRP. Eur J Cancer. 1996, 32A: 945-957. 10.1016/0959-8049(96)00046-9.CrossRefPubMed

13.

Venkatakrishnan ASA, King G, Blackwell TR, Bringham KL, Christman JW, Blackwell TS: Exaggerated activation of NF-kB and altered IkB-beta processing in cystic fibrosis bronchial epithelial cells. American Journal of Respiratory Cell Molecular Biology. 2000, 23: 396-403.CrossRefPubMed

14.

Henrion A, Flamant C, Roussey M, Strange RC, Chadelat K, Clement A: Severity of cystic fibrosis is associated with gene polymorphism of anti-oxidizing enzymes [abstract]. Pediatr Pulmonol. 2001, 22: s215-

15.

Baranov VS, Bakay B, Aseev M, Belotserkovskaya R, Baranova H, Malet P, Perriot J, Mouraire P, Baskakov VN, Savitskyi GA, Gorbushin S, Deyneka SI, Michnin E, Barchuck A, Vakharlovsky V, Pavlov G, Shilko VI, Guembitzkaya T, Kovaleva L: Proportion of the GSTM1 0/0 genotype in some Slavic populations and its correlation with cystic fibrosis and some multifactorial diseases. Hum Genet. 1996, 97: 516-520. 10.1007/BF02267078.CrossRefPubMed

16.

Jirsch JR, Deeley G: Inwardly rectifying K+ channels and volume-regulated anion channels in multidrug-resistant small cell lung cancer cells. Cancer Res. 1993, 53: 4156-4160.PubMed

17.

Hainsworth AH, Henderson RM: Hypotonicity-induced anion fluxes in cells expressing the multidrug-resistance-associated protein, MRP. Pflugers Arch. 1996, 432: 234-240. 10.1007/s004240050129.CrossRefPubMed

18.

Borst P, Kool RE, Wijnholds M: A family of drug transporters: the multidrug resistance-associated proteins. J Natl Cancer Inst. 2000, 92: 1295-1302. 10.1093/jnci/92.16.1295.CrossRefPubMed

19.

Leslie EM, Cole SP: Toxicological relevance of the multidrug resistance protein 1, MRP1 (ABCC1) and related transporters. Toxicology. 2001, 67: 3-23. 10.1016/S0300-483X(01)00454-1.CrossRef

20.

Wijnholds J, van Leudsen MR, Mol CA, Zaman GJ, Mayer U, Beijnen J, der Valk M, Krimpenfort P, Borst P: Increased sensitivity to anticancer drugs and decreased inflammatory response in mice lacking the multidrug resistance-associated protein. Nat Med. 1997, 3: 1275-1279. 10.1038/nm1197-1275.CrossRefPubMed

21.

Miller SA, Dykes DD: A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res. 1988, 16: 1215-CrossRefPubMedPubMedCentral

22.

Cipolli M, Novelli G, Cassetta MI, Fallani S, Mazzei T: Azithromycin concentrations in serum and bronchial secretions of patients with cystic fibrosis. Clin Drug Invest. 2001, 21: 353-10.2165/00044011-200121050-00005.CrossRef

23.

Wang Z, Wang B, Tang K, Lee EJD, Chong SS, Lee CGL: A functional polymorphism within the MRP1 gene locus identified through its genomic signature of positive selection. Hum Mol Gen. 2005, 14: 2075-2087. 10.1093/hmg/ddi212.CrossRefPubMed

24.

Zaldivar F, Nugent DJ: Identification of a novel regulatory element in the human interleukin 1 alpha (IL-1alpha) gene promoter. Cytokine. 2002, 20: 130-135. 10.1006/cyto.2002.1990.CrossRefPubMed

25.

Slapak CA, Fracasso PM, Martell RL, Toppmeyer DL, Lecerf JM, Levy SB: Overexpression of the multidrug resistance-associated protein (MRP) gene in vincristine but not doxorubicin-selected multidrug-resistant murine erythroleukemia cells. Cancer Res. 1994, 54: 5607-5613.PubMed

The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2350/7/7/prepub

Title: The GCC repeat length in the 5'UTR of MRP1 gene is polymorphic: a functional characterization of its relevance for cystic fibrosis
Authors: Elena Nicolis
Matteo Pasetto
Cristina Cigana
Ugo Pradal
Baroukh M Assael
Paola Melotti
Publication date: 01-12-2006
Publisher: BioMed Central
Published in: BMC Medical Genetics / Issue 1/2006
Electronic ISSN: 1471-2350
DOI: https://doi.org/10.1186/1471-2350-7-7

Keynote webinar | Spotlight on medication adherence

Springer Medicine

The GCC repeat length in the 5'UTR of MRP1 gene is polymorphic: a functional characterization of its relevance for cystic fibrosis

Abstract

Background

Methods

Results

Conclusion

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Background

Methods

Results

Conclusion

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