Reproductive Medical Center, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China.
Henan Key Laboratory of Reproduction and Genetics, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China.
Cell Prolif. 2020 Sep;53(9):e12876. doi: 10.1111/cpr.12876. Epub 2020 Aug 13.
NCOA6 is a transcription coactivator; its deletion in mice results in growth retardation and lethality between 8.5 and 12.5 dpc with defects in the placenta. However, the transcription factor(s) and the mechanism(s) involved in the function of NCOA6 in placentation have not been elucidated. Here, the roles of NCOA6 in human cytotrophoblast invasion and migration were studied.
Human placenta tissues were collected from normal pregnancies and pregnancies complicated by early-onset severe preeclampsia (sPE). Immunofluorescence, RT-qPCR and Western blotting were used to determine NCOA6 expression. Transwell invasion/migration assays were performed to explore whether NCOA6 knockdown affected human placenta-derived HTR-8/SVneo cell invasion/migration. Gelatin zymography was performed to examine the change in the gelatinolytic activities of secreted MMP2 and MMP9. Luciferase reporter assays were used to explore whether NCOA6 coactivated NF-κB-mediated MMP9 transcription.
NCOA6 is mainly expressed in the human placental trophoblast column, as well as in the EVTs. HTR-8/SVneo cell invasion and migration were significantly attenuated after NCOA6 knockdown, and the secretion of MMP9 was decreased due to transcriptional suppression. NCOA6 was further found to coactivate NF-κB-mediated MMP9 transcription. Moreover, expression of NCOA6 was impaired in placentas of patients complicated by early-onset sPE.
Thus, we demonstrated that NCOA6 is important for cytotrophoblast invasion/migration, at least partially, by activating NF-κB-mediated MMP9 transcription; the downregulation of NCOA6 may contribute to the pathogenesis of early-onset sPE.
NCOA6 是一种转录共激活因子；其在小鼠中的缺失导致 8.5 至 12.5 天胚泡期生长迟缓和致死，胎盘有缺陷。然而，参与 NCOA6 在胎盘形成中功能的转录因子（和）及其机制尚未阐明。本研究旨在研究 NCOA6 在人滋养细胞侵袭和迁移中的作用。
收集正常妊娠和早发性重度子痫前期（sPE）患者的胎盘组织。免疫荧光、RT-qPCR 和 Western blot 用于检测 NCOA6 的表达。Transwell 侵袭/迁移实验用于研究 NCOA6 敲低是否影响人胎盘来源的 HTR-8/SVneo 细胞侵袭/迁移。明胶酶谱法用于检测分泌型 MMP2 和 MMP9 的明胶酶活性变化。荧光素酶报告实验用于研究 NCOA6 是否共激活 NF-κB 介导的 MMP9 转录。
NCOA6 主要在人胎盘滋养细胞柱和 EVT 中表达。NCOA6 敲低后 HTR-8/SVneo 细胞侵袭和迁移明显减弱，由于转录抑制，MMP9 的分泌减少。进一步发现 NCOA6 共激活 NF-κB 介导的 MMP9 转录。此外，早发性 sPE 患者的胎盘表达 NCOA6 受损。
因此，我们证明 NCOA6 通过激活 NF-κB 介导的 MMP9 转录，对滋养细胞侵袭/迁移很重要；NCOA6 的下调可能导致早发性 sPE 的发病机制。
