Expression of P-glycoprotein in Human Placenta: Relation to Genetic Polymorphism of the Multidrug Resistance (MDR)-1 Gene

TLDR

Although the influence of these SNPs on the pharmacokinetics of P‑glycoprotein substrates remains unclear, the MDR‑1 polymorphisms identified here may inform future in vivo studies. The study aimed to determine whether MDR‑1 gene mutations correlate with placental P‑glycoprotein expression by sequencing cDNA and measuring protein levels in 100 placentas from Japanese women. Researchers developed PCR‑RFLP genotyping assays, sequenced MDR‑1 cDNA, and quantified P‑glycoprotein via Western blotting to assess mutation–expression relationships. Nine SNPs were detected, with G2677A/T and C3435T alleles strongly linked and associated with lower P‑glycoprotein expression; PCR‑RFLP proved reliable, mutation frequencies matched between cDNA and genomic DNA, and ethnic differences in polymorphism rates were suggested.

Abstract

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.