目的:在过去的四十年中,辅助生殖技术(ART)领域取得了许多进步。尽管如此,植入失败仍然是成功妊娠的关键限制步骤.子宫内膜容受性(ER)的建立对于成功植入至关重要。然而,ER的基本生物学过程和机制仍然难以捉摸。我们的研究调查了缺氧诱导因子-1α(HIF-1α)在ER建立过程中的功能,并阐明了HIF-1α调节ER相关基因表达网络的新分子机制。
方法:HIF-1α水平,homeoboxA10(HOXA10),胰岛素样生长因子结合蛋白1(IGFBP1),丙酮酸激酶M2(PKM2),实时荧光定量PCR检测子宫内膜组织中乳酸脱氢酶A(LDHA),免疫印迹和免疫组织化学。分别分析HIF-1α与HOXA10、IGFBP1、PKM2、LDHA的相关性。用载体HIF-1α处理Ishikawa细胞,HIF-1α-siRNA,和PKM2-siRNA。转染后,HOXA10,IGFBP1,LDHA,通过实时PCR和免疫印迹法检测PKM2,测定Ishikawa细胞的乳酸浓度和细胞迁移能力。
结果:HIF-1α水平,IGFBP1,HOXA10,LDHA,复发性种植失败(RIF)患者中PKM2明显下降,子宫内膜中HOXA10,IGFBP1,PKM2和LDHA的水平与HIF-1α相关。然后在HIF-1α载体和HIF-1α-siRNA建立的细胞模型中,HOXA10、IGFBP1、LDHA、PKM2和乳酸浓度显著上调和下调。PKM2-siRNA可显著降低HOXA10和IGFBP1的表达。
结论:HIF-1α通过调节糖酵解在ER的构建中起着至关重要的作用。
OBJECTIVE: Numerous advancements have been introduced into the field of assisted reproductive technology (ART) in the past four decades. Nonetheless, implantation failure is still a key limiting step for a successful pregnancy. Building of endometrial receptivity (ER) is essential for successful implantation. However, the fundamental biological processes and mechanisms of ER remain elusive. Our study investigates the function of hypoxia inducible factor-1α (HIF-1α) during ER establishment and shed lights on the novel molecular mechanism by which HIF-1α regulates ER-related gene expression network.
METHODS: Levels of HIF-1α, homeobox A10 (HOXA10), insulin-like growth factor-binding protein 1 (IGFBP1), pyruvate kinase M2 (PKM2), and lactate dehydrogenase A (LDHA) in endometrial tissues were measured via real-time PCR, immunoblotting and immunohistochemistry. The correlation between HIF-1α and HOXA10, IGFBP1, PKM2, LDHA were analyzed separately. Ishikawa cells were treated with vector HIF-1α, HIF-1α-siRNA, and PKM2-siRNA. After transfection, the levels of HOXA10, IGFBP1, LDHA, and PKM2 were measured via real-time PCR and immunoblotting, and the lactate concentrations and cell migration of Ishikawa cells were measured.
RESULTS: Levels of HIF-1α, IGFBP1, HOXA10, LDHA, and PKM2 were significantly decreased in recurrent implantation failure (RIF) patients and levels of HOXA10, IGFBP1, PKM2, and LDHA were correlated with HIF-1α in endometrium. Then in a cellular model established by HIF-1α vector and HIF-1α-siRNA, the expression of HOXA10, IGFBP1, LDHA, PKM2, and lactate concentrations were dramatically upregulated and downregulated. And the expression of HOXA10, and IGFBP1 were dramatically decreased by PKM2-siRNA.
CONCLUSIONS: HIF-1α plays a crucial role in the building of ER through regulating glycolysis.