Key Laboratory of Reproduction Regulation of NHFPC, Shanghai Institute of Planned Parenthood Research, Pharmacy School, Fudan University, Shanghai, China.
The Second Hospital of Tianjin Medical University, Tianjin, China.
J Transl Med. 2018 Jul 4;16(1):186. doi: 10.1186/s12967-018-1556-x.
Implantation failure is not only a major cause of early pregnancy loss, but it is also an obstacle to assisted reproductive technologies. The identification of potential circulating biomarkers for recurrent miscarriage (RM) and/or recurrent implantation failure would contribute to the development of novel diagnosis and prediction techniques.
MiR (miR-23a-3p, 27a-3p, 29a-3p, 100-5p, 127-3p and 486-5p) expression in the villi, decidual tissues and peripheral blood plasma and serum were validated by qPCR, and the localization of miRs in the villi and decidual tissues of RM and normal pregnancy (NP) women were detected by in situ hybridization. The invasiveness of HTR8/SVneo cells was determined using a Transwell assay. The predictive values of miRs for RM and the outcome of IVF-ET were respectively calculated by the receiver operating characteristic analysis.
The signals of six miRs were observed in the villi and decidual tissues of RM and NP women. The villus miR-27a-3p, miR-29a-3p and miR-100-5p were significantly up-regulated, whereas miR-127-3p and miR-486-5p appeared to be down-regulated in RM women compared to NP women. The invasiveness of HTR8/SVneo cells transfected with miR-23a-3p mimics was evidently weakened, whereas that of cells transfected with miR-127-3p mimics was obviously enhanced. The peripheral blood plasma levels of miR-27a-3p, miR-29a-3p, miR-100-5p and miR-127-3p were significantly increased, whereas that of miR-486-5p was remarkably decreased in RM compared to NP women. By contrast, serum miR-23a-3p and miR-127-3p were significantly decreased, whereas that of miR-486-5p was remarkably increased. The combination of six plasma miRs levels discriminated RM with a sensitivity of 100% and a specificity of 83.3%, whereas that of six serum miRs levels showed a sensitivity of 78.3% and a specificity of 93.1%. In the IVF-ET cohort, the significantly decreased peripheral blood plasma levels of miR-23a-3p, miR-27a-3p, miR-100-5p and miR-127-3p, and the serum levels of miR-100-5p and miR-486-5p, in addition to the significantly increased serum level of miR-27a-3p, were found to be associated with the failure of ET. Moreover, the combination of plasma miR-23a-3p, miR-27a-3p, miR-29a-3p, miR-100-5p, miR-127-3p and miR-486-5p levels discriminated the outcome of IVF-ET with a sensitivity of 68.1% and a specificity of 54.1%, whereas the combination of plasma miR-127-3p and miR-486-5p levels showed a sensitivity of 50.0% and a specificity of 75.3%.
Circulating miR-23a-3p, miR-27a-3p, miR-29a-3p, miR-100-5p, miR-127-3p and miR-486-5 might be involved in RM pathogenesis and present potential diagnostic biomarkers for RM. Meanwhile, these miRs, in particular miR-127-3p and miR-486-5p, provide promising prediction indexes for the outcomes of IVF-ET.
着床失败不仅是早期妊娠丢失的主要原因，也是辅助生殖技术的障碍。识别复发性流产（RM）和/或复发性着床失败的潜在循环生物标志物将有助于开发新的诊断和预测技术。
通过 qPCR 验证了 miR（miR-23a-3p、27a-3p、29a-3p、100-5p、127-3p 和 486-5p）在绒毛、蜕膜组织和外周血浆及血清中的表达，并通过原位杂交检测了 miR 在 RM 和正常妊娠（NP）妇女绒毛和蜕膜组织中的定位。使用 Transwell 测定 HTR8/SVneo 细胞的侵袭性。通过接收者操作特征分析分别计算 miR 对 RM 的预测值和 IVF-ET 的结果。
在 RM 和 NP 妇女的绒毛和蜕膜组织中观察到六种 miR 的信号。与 NP 妇女相比，RM 妇女的绒毛 miR-27a-3p、miR-29a-3p 和 miR-100-5p 明显上调，而 miR-127-3p 和 miR-486-5p 似乎下调。转染 miR-23a-3p 模拟物的 HTR8/SVneo 细胞的侵袭性明显减弱，而转染 miR-127-3p 模拟物的细胞的侵袭性明显增强。与 NP 妇女相比，RM 妇女的外周血浆 miR-27a-3p、miR-29a-3p、miR-100-5p 和 miR-127-3p 水平显著升高，而 miR-486-5p 水平明显降低。相比之下，血清 miR-23a-3p 和 miR-127-3p 明显降低，而 miR-486-5p 明显升高。六种血浆 miR 水平的组合对 RM 的灵敏度为 100%，特异性为 83.3%，而六种血清 miR 水平的组合对 RM 的灵敏度为 78.3%，特异性为 93.1%。在 IVF-ET 队列中，明显降低的外周血浆 miR-23a-3p、miR-27a-3p、miR-100-5p 和 miR-127-3p 水平以及血清 miR-100-5p 和 miR-486-5p 水平，以及血清 miR-27a-3p 水平的明显升高，与 ET 失败有关。此外，血浆 miR-23a-3p、miR-27a-3p、miR-29a-3p、miR-100-5p、miR-127-3p 和 miR-486-5p 水平的组合对 IVF-ET 的结果具有 68.1%的灵敏度和 54.1%的特异性，而血浆 miR-127-3p 和 miR-486-5p 水平的组合对 IVF-ET 的结果具有 50.0%的灵敏度和 75.3%的特异性。
循环 miR-23a-3p、miR-27a-3p、miR-29a-3p、miR-100-5p、miR-127-3p 和 miR-486-5 可能参与 RM 的发病机制，并为 RM 提供潜在的诊断生物标志物。同时，这些 miR，特别是 miR-127-3p 和 miR-486-5p，为 IVF-ET 的结果提供了有前途的预测指标。
