Single nucleotide polymorphisms in premature ovarian failure-associated genes in a Chinese Hui population

Abstract

Premature ovarian failure (POF) is an ovarian defect characterized by the premature depletion of ovarian follicles in individuals <40 years old, and is a major cause of infertility in females. Genetic factors are considered to be responsible for the development of POF, however, the exact pathogenesis remains to be elucidated in the majority of cases. In the present study, the single nucleotide polymorphisms (SNPs) of growth differentiation factor 9 (GDF9), bone morphogenetic protein 15 (BMP15), inhibin βB (INHBB) and follicle stimulating hormone receptor (FSHR) genes were investigated, and their association with POF in a Chinese Hui population of the Ningxia Hui Autonomous Region in western China was evaluated. Peripheral blood samples were collected from 63 patients diagnosed with POF (POF group) and 58 normal control individuals (control group), from which the genomic DNA was isolated. The GDF9, BMP15, INHBB and FSHR genes were amplified using polymerase chain reaction assays, and their SNPs were determined by sequencing. In the four SNPs identified across the GDF9 loci, D57Y (169G>T), rs1049127 (546G>A), rs254286 (447C>T) and rs254285 (969C>G), the frequencies of the 546G>A genotype and allele A were significantly higher in the POF group, compared with the normal control group (34.92, vs. 6.90%; P<0.05 and 19.05, vs. 3.23%; P<0.05, respectively), while no significant differences were observed in the occurrence of the c.447C>T and c.969C>G mutations between the two groups (60.32, vs. 50% and 50.79, vs. 55.17%, respectively). The c.169G>T mutation within the GDF9 gene was only detected in two patients with POF, and the mutation did not occur in the normal control group. A total of three SNPs were detected within the BMP15 gene, including rs3810682 (-9C>G), rs79377927 (788_789insTCT) and rs17003221 (852C>T), and no significant differences were observed in the frequencies of the -9C>G and 852C>T genotypes between the POF and control groups (7.94, vs. 6.90% and 4.76, vs. 3.45%, respectively). The 788_789insTCT genotype was detected in only two patients with POF. A novel mutation, c.1095C>A, was identified in exon 2 of the INHBB gene, however, no significant difference was found in the occurrence of the mutation between the two groups (30.16, vs. 22.41%; P>0.05). The rs6165 (919G>A) and rs6166 (2039G>A) SNPs were detected in exon 10 of the FSHR gene; however, no significant difference was observed in the genotype frequencies between the two groups (92.06, vs. 91.38% and 96.83, vs. 93.10%, respectively). These results demonstrated that GDF9 c.169G>T (D57Y), c.546G>A (rs1049127), and BMP15 rs79377927 (788_789insTCT) were associated with POF in the Chinese Hui population.

Figure 1

Sequencing map of GDF9 169G>T. (A) GG wild type genotype; (B) GT heterozygote. GDF9, growth differentiation factor 9.

Figure 2

Sequencing map of the 546G>T growth differentiation factor 9 gene mutation. (A) GG wild type genotype; (B) GT heterozygote; (C) TT homozygous mutation.

Figure 3

Sequencing map of the 448C>T growth differentiation factor 9 mutation. (A) CC wild type genotype; (B) CT heterozygote; (C) TT homozygous mutation. GDF9, growth differentiation factor 9.

Figure 4

Sequencing map of GDF9 969C>G. (A) CC wild type genotype; (B) CG heterozygote; (C) GG homozygous mutation. GDF9, growth differentiation factor 9.

Figure 5

Sequencing map of the rs79377927 (788_789insTCT) bone morphogenetic protein 15. (A) wild type genotype; (B) insTCT.

Figure 6

Sequencing map of the −9C>G bone morphogenetic protein 15 mutation. (A) CC wild type genotype; (B) CG heterozygote; (C) GG homozygous mutation.

Figure 7

Sequencing map of the 852C>T bone morphogenetic protein 15 mutation. (A) CC wild type genotype; (B) CT heterozygote.

Figure 8

Sequencing map of the 1095C>A inhibin B mutation. (A) CC wild type genotype; (B) CA heterozygote.

Figure 9

Sequencing map of the 919G>A follicle stimulating hormone receptor gene. (A) GG wild type genotype; (B) GA heterozygote; (C) AA homozygous mutation.

Figure 10

Sequencing map of the 2039G>A follicle stimulating hormone receptor.gene mutation. (A) GG wild type genotype; (B) GA heterozygote; (C) AA homozygous mutation.