Yang Yang, Liu Sai, Zhu Xiao-Ming, Chen You-Yi, Zhao Jing, Yuan Yu-Fei, Ma Yuan
Reproductive Medicine Center, Xi'an People's Hospital (Xi'an Fourth Hospital), Xi'an, P. R. China.
Department of Obstetrics and Gynecology, Hainan Branch of PLA General Hospital, Sanya, P. R. China.
Kaohsiung J Med Sci. 2025 May;41(5):e70019. doi: 10.1002/kjm2.70019. Epub 2025 May 1.
Trophoblast cells are specialized placental epithelial cells essential for pregnancy maintenance. miR-152 is implicated in trophoblast cell regulation and pregnancy failure. This study explores the role of miR-152 in decidual natural killer (dNK) cell viability and trophoblast cell invasion. HTR-8/SVneo cells were transfected with miR-152-mimics/inhibitor or their respective controls, followed by co-culture with dNK cells. RT-qPCR assessed transfection efficiency, while cytokine secretion (IL-8, IP-10, VEGF), cell viability, apoptosis, and invasion were evaluated via ELISA, CCK-8, flow cytometry, Western blot, and Transwell assays. The interaction between miR-152 and HLA-G was examined via dual-luciferase reporter assay, and HLA-G/sHLA-G levels were measured. Co-cultures of dNK cells and miR-152/HLA-G-overexpressing HTR-8/SVneo cells were established, and anti-KIR2DL4/IgG1 was used to block HLA-G/KIR2DL4 binding. Co-immunoprecipitation confirmed protein interactions. miR-152 overexpression suppressed dNK cell cytokine secretion, reduced HTR-8/SVneo cell viability and invasion, and promoted apoptosis. miR-152 inhibition had the opposite effect. miR-152 directly targeted HLA-G, and HLA-G overexpression rescued dNK function and trophoblast invasion. Blocking the HLA-G/KIR2DL4 binding counteracted the effects of miR-152. miR-152 inhibits dNK cell function and trophoblast invasion by targeting HLA-G, reducing HLA-G/KIR2DL4 interaction. These findings highlight a potential regulatory mechanism in pregnancy maintenance.
滋养层细胞是维持妊娠所必需的特殊胎盘上皮细胞。miR - 152与滋养层细胞调节及妊娠失败有关。本研究探讨miR - 152在蜕膜自然杀伤(dNK)细胞活力和滋养层细胞侵袭中的作用。将miR - 152模拟物/抑制剂或其各自的对照转染至HTR - 8/SVneo细胞,随后与dNK细胞共培养。通过RT - qPCR评估转染效率,而细胞因子分泌(IL - 8、IP - 10、VEGF)、细胞活力、凋亡和侵袭则通过ELISA、CCK - 8、流式细胞术、蛋白质印迹法和Transwell试验进行评估。通过双荧光素酶报告基因试验检测miR - 152与HLA - G之间的相互作用,并测量HLA - G/sHLA - G水平。建立dNK细胞与过表达miR - 152/HLA - G的HTR - 8/SVneo细胞的共培养体系,并使用抗KIR2DL4/IgG1阻断HLA - G/KIR2DL4结合。免疫共沉淀证实了蛋白质相互作用。miR - 152过表达抑制dNK细胞因子分泌,降低HTR - 8/SVneo细胞活力和侵袭,并促进凋亡。miR - 152抑制则产生相反的效果。miR - 152直接靶向HLA - G,HLA - G过表达可挽救dNK功能和滋养层侵袭。阻断HLA - G/KIR2DL4结合可抵消miR - 152的作用。miR - 152通过靶向HLA - G抑制dNK细胞功能和滋养层侵袭,减少HLA - G/KIR2DL4相互作用。这些发现突出了妊娠维持中的一种潜在调节机制。