Center for Reproductive Medicine, Henan Key Laboratory of Reproduction and Genetics, The First Affiliated Hospital of Zhengzhou University, 40, Daxue Road, Zhengzhou, Henan, China.
Reprod Biol Endocrinol. 2022 Jan 31;20(1):22. doi: 10.1186/s12958-022-00902-9.
Tightly regulation of extravillous cytotrophoblast (EVT) cell invasion is critical for the placentation and establishment of a successful pregnancy. Insufficient EVT cell invasion leads to the development of preeclampsia (PE) which is a leading cause of maternal and perinatal mortality and morbidity. Transforming growth factor-beta1 (TGF-β1) and kisspeptin are expressed in the human placenta and have been shown to inhibit EVT cell invasion. Kisspeptin is a downstream target of TGF-β1 in human breast cancer cells. However, whether kisspeptin is regulated by TGF-β1 and mediates TGF-β1-suppressed human EVT cell invasion remains unclear.
The effect of TGF-β1 on kisspeptin expression and the underlying mechanisms were explored by a series of in vitro experiments in a human EVT cell line, HTR-8/SVneo, and primary cultures of human EVT cells. Serum levels of TGF-β1 and kisspeptin in patients with or without PE were measured by ELISA.
TGF-β1 upregulates kisspeptin expression in HTR-8/SVneo cells and primary cultures of human EVT cells. Using pharmacological inhibitor and siRNA, we demonstrate that the stimulatory effect of TGF-β1 on kisspeptin expression is mediated via the ALK5 receptor. Treatment with TGF-β1 activates SMAD2/3 canonical pathways as well as ERK1/2 and PI3K/AKT non-canonical pathways. However, only inhibition of ERK1/2 activation attenuates the stimulatory effect of TGF-β1 on kisspeptin expression. In addition, siRNA-mediated knockdown of kisspeptin attenuated TGF-β1-suppressed EVT cell invasion. Moreover, we report that serum levels of TGF-β1 and kisspeptin are significantly upregulated in patients with PE.
By illustrating the potential physiological role of TGF-β1 in the regulation of kisspeptin expression, our results may serve to improve current strategies used to treat placental diseases.
滋养细胞外突（EVT）细胞侵袭的严格调控对于胎盘形成和成功妊娠的建立至关重要。EVT 细胞侵袭不足会导致子痫前期（PE）的发生，这是孕产妇和围产儿发病率和死亡率的主要原因。转化生长因子-β1（TGF-β1）和 kisspeptin 在人胎盘组织中表达，并已被证明可抑制 EVT 细胞侵袭。Kisspeptin 是人类乳腺癌细胞中 TGF-β1 的下游靶标。然而，kisspeptin 是否受 TGF-β1 调控并介导 TGF-β1 抑制的人 EVT 细胞侵袭尚不清楚。
通过一系列体外实验，在人 EVT 细胞系 HTR-8/SVneo 和人 EVT 细胞原代培养物中探讨了 TGF-β1 对 kisspeptin 表达的影响及其潜在机制。通过 ELISA 法测定有或无 PE 的患者血清中 TGF-β1 和 kisspeptin 的水平。
TGF-β1 可上调 HTR-8/SVneo 细胞和人 EVT 细胞原代培养物中的 kisspeptin 表达。使用药理学抑制剂和 siRNA，我们证明 TGF-β1 对 kisspeptin 表达的刺激作用是通过 ALK5 受体介导的。TGF-β1 处理可激活 SMAD2/3 经典途径以及 ERK1/2 和 PI3K/AKT 非经典途径。然而，只有抑制 ERK1/2 激活才能减弱 TGF-β1 对 kisspeptin 表达的刺激作用。此外，siRNA 介导的 kisspeptin 敲低可减弱 TGF-β1 抑制的 EVT 细胞侵袭。此外，我们报告在 PE 患者中血清 TGF-β1 和 kisspeptin 水平显著上调。
通过阐明 TGF-β1 在调节 kisspeptin 表达中的潜在生理作用，我们的研究结果可能有助于改善目前用于治疗胎盘疾病的策略。
