Department of Infectious Diseases, Institute for Viral Hepatitis, Key Laboratory of Molecular Biology for Infectious Diseases (Ministry of Education), The Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, P.R. China.
Department of Gynecology and Obstetrics, The Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, P.R. China.
Mol Med Rep. 2018 Mar;17(3):3481-3488. doi: 10.3892/mmr.2017.8338. Epub 2017 Dec 22.
Despite the use of adjuvant therapies, the cumulative proportion of live births remains at ~40%. Accumulating data show that low pregnancy rates, even in the presence of high fertility rates, are due to implantation failure. The present study aimed to identify and construct a profile of proteins that react with preimplantation factor (PIF) and to provide an understanding into the molecular mechanisms by which PIF promotes trophoblast invasion. Cytoplasmic proteins were immunoprecipitated with biotin‑labeled synthetic PIF or intralipid and scrambled PIF (PIFscr). The protein profiles were analyzed using isobaric tags for relative and absolute quantification coupled with mass spectrometry. Immunoprecipitation and western blot analyses were used to assess the interactions between PIF and myosin heavy chain 10 (MYH10) and heat shock protein family D1. Small interfering RNA‑based silencing was performed to examine the function of MYH10. In the results of the present study, 21 proteins were identified with interactions with PIF. The immunoprecipitation and western blot analyses revealed an interaction between PIF and MYH10. Silencing of the expression of MYH10 in HEC‑1‑B cells significantly attenuated cell migration and invasion capacities. These data support the conclusion that MYH10‑mediated cell migration and invasion act in conjunction with PIF to promote the trophoblast invasion procedure.
尽管使用了辅助治疗方法，但活产儿的累积比例仍约为 40%。累积数据表明，即使生育率高，妊娠率仍然较低，这是由于着床失败所致。本研究旨在鉴定和构建与着床前因子（PIF）反应的蛋白质谱，并深入了解 PIF 促进滋养层侵袭的分子机制。用生物素标记的合成 PIF 或脂内液和乱序 PIF（PIFscr）对细胞质蛋白进行免疫沉淀。使用等重标记相对和绝对定量结合质谱法分析蛋白质谱。免疫沉淀和 Western blot 分析用于评估 PIF 与肌球蛋白重链 10（MYH10）和热休克蛋白家族 D1 之间的相互作用。基于小干扰 RNA 的沉默用于研究 MYH10 的功能。在本研究的结果中，鉴定出 21 种与 PIF 相互作用的蛋白质。免疫沉淀和 Western blot 分析显示 PIF 与 MYH10 之间存在相互作用。在 HEC-1-B 细胞中沉默 MYH10 的表达显著减弱了细胞迁移和侵袭能力。这些数据支持 MYH10 介导的细胞迁移和侵袭与 PIF 共同作用以促进滋养层侵袭过程的结论。
