Reproductive Medical Center, Renmin Hospital of Wuhan University and Hubei Clinic Research Center for Assisted Reproductive Technology and Embryonic Development, Wuhan, Hubei, China.
Department of Spine Surgery & Musculoskeletal Tumor, Zhongnan Hospital of Wuhan University, Wuhan, Hubei, China.
J Reprod Immunol. 2022 Aug;152:103647. doi: 10.1016/j.jri.2022.103647. Epub 2022 May 23.
The purpose of this study was to determine the regulation of the miR-410-5p /ITGA6 axis on the biological functions of trophoblast cells and the mechanism involved in recurrent spontaneous abortion(RSA). We used qRT-PCR and Western blotting to quantify the expression levels of Mir-410-5p and ITGA6 in placenta of RSA and normal, and found that compared with normal placenta, the placenta of RSA patients showed higher miR-410-5p and lower ITGA6 expression. Dual luciferase reporter gene assay confirmed the binding of miR-410-5p to ITGA6. The expression of miR-410-5p and ITGA6, and proliferation, apoptosis, invasion and migration of trophoblast cells and the effect on the polarization of M2 macrophages were detected in the trophoblast derived cell lines HTR8/Svneo transfected with miR-410-5p mimic, sh-miR-410-5p and si-ITGA6 respectively. Meanwhile, the molecular mechanism of ITGA6 regulation on trophoblast cells was explored. Transfection with miR-410-5p mimic or si-ITGA6 attenuated the proliferation, migration and invasion and induced apoptosis of HTR-8/SVneo cells. Transfection of sh-miR-410-5p promoted proliferation, migration and invasion, and weakened apoptosis of HTR-8/SVneo cells. In addition, overexpression of miR-410-5p in trophoblast cells inhibited the polarization of M2 macrophages, while knockdown of miR-410-5p was beneficial to recruitment of trophoblast cell and promoted the polarization of M2 macrophages. ITGA6 may affect the biological functions of trophoblast cells by regulating PI3K/AKT and MAPK signaling pathways. In conclusion, miR-410-5p mediates trophoblast cell proliferation, apoptosis, invasion and migration through regulating ITGA6 expression.
本研究旨在确定 miR-410-5p/ITGA6 轴对滋养层细胞生物学功能的调节作用及其在复发性自然流产(RSA)中的作用机制。我们使用 qRT-PCR 和 Western blot 技术定量检测了 RSA 和正常胎盘组织中 Mir-410-5p 和 ITGA6 的表达水平,发现与正常胎盘相比,RSA 患者胎盘组织中 miR-410-5p 表达升高,ITGA6 表达降低。双荧光素酶报告基因实验证实了 miR-410-5p 与 ITGA6 的结合。分别转染 miR-410-5p 模拟物、sh-miR-410-5p 和 si-ITGA6 后,检测滋养层来源细胞系 HTR8/Svneo 中 miR-410-5p 和 ITGA6 的表达以及滋养层细胞的增殖、凋亡、侵袭和迁移,并观察对 M2 巨噬细胞极化的影响。同时,探讨了 ITGA6 对滋养层细胞的调控作用的分子机制。转染 miR-410-5p 模拟物或 si-ITGA6 可减弱 HTR-8/SVneo 细胞的增殖、迁移和侵袭,诱导细胞凋亡。转染 sh-miR-410-5p 则促进 HTR-8/SVneo 细胞的增殖、迁移和侵袭,减弱细胞凋亡。此外,在滋养层细胞中过表达 miR-410-5p 可抑制 M2 巨噬细胞的极化,而敲低 miR-410-5p 则有利于滋养层细胞的募集,并促进 M2 巨噬细胞的极化。ITGA6 可能通过调节 PI3K/AKT 和 MAPK 信号通路影响滋养层细胞的生物学功能。综上所述,miR-410-5p 通过调节 ITGA6 的表达来介导滋养层细胞的增殖、凋亡、侵袭和迁移。