Department of Obstetrics and Gynecology, Shengjing Hospital Affiliated to China Medical University, Shenyang, Liaoning 110000, P.R. China.
Int J Mol Med. 2018 May;41(5):2927-2934. doi: 10.3892/ijmm.2018.3503. Epub 2018 Feb 16.
Sex hormone-binding globulin (SHBG) has a key role in the occurrence and development of the gestational diab-etes mellitus (GDM). Single nucleotide polymorphism (SNP) rs6259 is a functional site in SHBG gene, which is suspected to regulate the SHBG level. The present study explored the placental SHBG SNP rs6259 distribution in Chinese pregnant women and the influence on placental SHBG concentrations, to assess the relationship of SHBG rs6259 in the occurrence and development of GDM. We screened the SHBG rs6259 allele in 210 healthy and 180 GDM gravidas by PCR-RFLP and restriction enzyme and measured placental SHBG concentrations in each genotypic group with western blot analysis. The mechanisms of SHBG rs6259 function were analyzed by cell culture, recombinant lentivirus transfection, real-time PCR, and western blot analysis. We found the differences of SHBG Asn327 allele frequency and the genotype distribution in GDM and control groups were statistically significant (P<0.05). Western blot analysis results showed that the Asn327 allele group was associated with a higher placental SHBG level than the Asp327 allele homozygote group (P<0.05). In HTR8-SVneo cell transfection, the positive transfection groups (SHBG-rs6259 Asn) led to an obviously higher tendency of SHBG mRNA and protein expression than the negative control groups (SHBG-rs6259 Asp), the normal cell group, and the blank control group (blank lentivirus LV-5) (P<0.05). Our data, therefore, reflected that SHBG SNP rs6259 causes changes in placental SHBG concentration and may play a functional role in the molecular mechanisms of GDM etiology.
性激素结合球蛋白（SHBG）在妊娠期糖尿病（GDM）的发生和发展中起着关键作用。单核苷酸多态性（SNP）rs6259 是 SHBG 基因中的一个功能位点，据推测它可以调节 SHBG 水平。本研究探讨了中国孕妇胎盘 SHBG SNP rs6259 的分布及其对胎盘 SHBG 浓度的影响，以评估 SHBG rs6259 与 GDM 发生发展的关系。我们通过 PCR-RFLP 和限制性内切酶筛选了 210 例健康孕妇和 180 例 GDM 孕妇的 SHBG rs6259 等位基因，并通过 Western blot 分析测定了每个基因型组的胎盘 SHBG 浓度。通过细胞培养、重组慢病毒转染、实时 PCR 和 Western blot 分析分析了 SHBG rs6259 功能的机制。我们发现，SHBG rs6259 Asn327 等位基因频率和基因型分布在 GDM 和对照组之间存在统计学差异（P<0.05）。Western blot 分析结果表明，Asn327 等位基因组与 Asp327 等位基因纯合子组相比，胎盘 SHBG 水平更高（P<0.05）。在 HTR8-SVneo 细胞转染中，阳性转染组（SHBG-rs6259 Asn）的 SHBG mRNA 和蛋白表达明显高于阴性对照组（SHBG-rs6259 Asp）、正常细胞组和空白对照组（空白慢病毒 LV-5）（P<0.05）。因此，我们的数据反映了 SHBG SNP rs6259 导致胎盘 SHBG 浓度的变化，并且可能在 GDM 病因学的分子机制中发挥功能作用。
