In vitro up-regulation of HLA-G using dexamethasone and hydrocortisone in first-trimester trophoblast cells of women experiencing recurrent miscarriage
In vitro up-regulation of HLA-G using dexamethasone and hydrocortisone in first-trimester trophoblast cells of women experiencing recurrent miscarriage
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DOI：
10.1111/j.1399-0039.2012.01884.x
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发表时间：
2012-08-01
期刊：
影响因子：
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通讯作者：
Agrawal, S.
中科院分区：
文献类型：
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作者：
Akhter, A.;Faridi, R. M.;Agrawal, S.
母胎界面的滋养细胞表达一种罕见的人类白细胞抗原(HLA)-C、HLA- e和HLA- g的组合。HLA-G在胞外滋养细胞上的表达改变与复发性流产（RMs）的病因有关。我们评估了从RM患者制备的细胞培养物中上皮外细胞滋养细胞中HLA-G的表达，并与妊娠早期自愿终止正常妊娠的患者（对照组）进行了比较。糖皮质激素、地塞米松和氢化可的松在调节HLA-G表达中的作用。研究了HLA-G启动子和3'UTR变异对HLA-G转录的影响。用地塞米松和氢化可的松（剂量浓度为01000 ng/ml）治疗妊娠早期RM患者培养的细胞滋养细胞。采用半定量和定量实时聚合酶链反应（RT-PCR）检测HLA-G基因转录，采用特异性酶联免疫吸附试验（ELISA）、流式细胞术和western blot检测蛋白表达。通过PCR和/或序列分型检测HLA-G多态性。与对照组相比，从RM患者获得的未经处理的滋养细胞中观察到低水平的HLA-G。经糖皮质激素处理后，这些细胞中HLA-G的表达呈剂量依赖性上调（P < 0.05），但细胞增殖和活力未发生变化。RM患者与对照组HLA-G多态性无显著相关性。HLA-G在RM患者培养的滋养细胞中表达最低。它可以在暴露于地塞米松和氢化可的松时上调。糖皮质激素在体外具有调节HLA-G表达的潜力，可以进一步研究其在RM中的治疗适用性。
The trophoblast cells at the maternalfetal interface express an unusual combination of human leukocyte antigen (HLA)-C, HLA-E and HLA-G. Altered expression of HLA-G on the extravillous cytotrophoblast has been implicated in the etiology of recurrent miscarriages (RMs). We have assessed HLA-G expression in extravillous cytotrophoblast in cell cultures prepared from RM patients and compared with those of first-trimester voluntarily terminated normal pregnancies (control). Glucocorticoids, dexamethasone and hydrocortisone were examined for their role in modulation of the HLA-G expression. HLA-G promoter and 3'UTR variants were investigated for their effect on the transcription of HLA-G. Cultured cytotrophoblast cells from the first-trimester RM patients were treated with dexamethasone and hydrocortisone (dose concentration 01000 ng/ml). HLA-G gene transcription was determined by semiquantitative and quantitative real-time polymerase chain reaction (RT-PCR), while protein expression was determined by a specific enzyme-linked immunosorbent assay (ELISA), flow cytometry and western blot analyses. HLA-G polymorphisms were detected by PCR and/or sequence-based typing. Low level of HLA-G was observed in untreated trophoblast cells obtained from RM patients as compared with controls. Upon treatment with glucocorticoids, the expression of HLA-G in these cells was up-regulated in a dose-dependent manner (P < 0.05), with no change in cellular proliferation and viability. There was no significant association between HLA-G polymorphism in RM patients and controls. HLA-G is minimally expressed in cultured trophoblast cells of RM patients. It can be up-regulated upon exposure with both dexamethasone and hydrocortisone. Glucocorticoids have the potential to modulate HLA-G expression in vitro, and can be further examined for their therapeutic applicability in RM.