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Vol. 297, Issue 3, 1137-1143, June 2001
Department of Clinical Pharmacokinetics, Division of Pharmaceutical
Sciences, Graduate School, Kyushu University, Fukuoka, Japan (Mi.T.,
S.I., Y.Ku., S.H.); Department of Hospital Pharmacy (I.I., K.I., K.O.)
and Department of Obstetrics and Gynecology (N.N., Y.Ka., Ma.T., J.K.,
N.T.), Faculty of Medicine, Yonago University, Tottori, Japan
To evaluate whether mutations in the human multidrug resistance (MDR)-1
gene correlate with placental P-glycoprotein (PGP) expression, we
sequenced the MDR-1 cDNA and measured PGP expression by Western
blotting in 100 placentas obtained from Japanese women. Nine single
nucleotide polymorphisms (SNPs) were observed with an allelic frequency
of 0.005 to 0.420. Of these SNPs, G2677A (allelic frequency = 0.18) and G2677T (0.39) in exon 21 were associated with an amino acid
conversion from Ala to Thr and to Ser, respectively. Sixty-one of 65 samples (93.8%), which had a C3435T allele, also had a mutant
G2677(A,T) allele, suggesting an association between the two SNPs.
Correlations of mutations with expression levels were observed;
individuals having the G2677(A,T) and/or T-129C (p < 0.05) allele had less placental PGP. Polymerase chain
reaction-restriction fragment length polymorphism (PCR-RFLP)-based
genotyping tests were developed for the detection of these SNPs. The
PCR, in which genomic DNAs obtained from healthy subjects
(n = 48) are used as samples, was successful. The
frequency of mutations in placental cDNA was identical with that in
genomic DNA. When genotype results were compared between Caucasians and
Japanese, ethnic differences in the frequency of polymorphism in the
MDR-1 gene were suspected. Although it remains to be determined whether
these SNPs influence the pharmacokinetic and dynamic properties of
clinically useful drugs that are substrates of PGP, the polymorphism of
the MDR-1 gene presented here may provide useful information in in vivo study of these issues.
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