Department of Biomedical Science, Cell and Gene Therapy Research Institute, CHA University, Seongnam 13488, Republic of Korea.
Department of Obstetrics and Gynecology, CL Women's Hospital, Gwangju 61917, Republic of Korea.
Cells. 2023 May 24;12(11):1459. doi: 10.3390/cells12111459.
The expression of mRNA is significantly lower in the chorionic villi of patients with recurrent pregnancy loss (RPL) than in the control group. We conducted an investigation into the cellular functions of HtrA4 using the CRISPR/Cas9 system and shRNA- to create knockout BeWo cells and knockdown JEG3 cells. Our results indicated that the knockout BeWo cells exhibited reduced capacity for invasion and fusion, but increased levels of proliferation and migration, with a significantly shortened cell cycle compared to wild-type cells. Wild-type BeWo cells highly expressed cell invasion- and fusion-related factors, while knockout BeWo cells highly expressed migration-, proliferation-, and cell cycle-related factors. The shRNA- JEG3 cells showed a decreased capacity for invasion, but an increased capacity for migration, accompanied by a decrease in the expression of cell invasion-related factors and an increase in migration-related factors. Moreover, our ELISA results revealed that the serum HtrA4 level was lower in patients with RPL than in the controls. These findings suggest that HtrA4 depletion may be associated with placental dysfunction.
在复发性流产（RPL）患者的绒毛中，mRNA 的表达明显低于对照组。我们使用 CRISPR/Cas9 系统和 shRNA 构建了 HtrA4 的敲除 BeWo 细胞和敲低 JEG3 细胞，研究了 HtrA4 的细胞功能。结果表明，与野生型细胞相比，敲除 BeWo 细胞的侵袭和融合能力降低，增殖和迁移能力增强，细胞周期明显缩短。野生型 BeWo 细胞高表达与侵袭和融合相关的因子，而敲除 BeWo 细胞高表达与迁移、增殖和细胞周期相关的因子。shRNA-JEG3 细胞的侵袭能力降低，但迁移能力增强，同时侵袭相关因子的表达降低，迁移相关因子的表达增加。此外，我们的 ELISA 结果显示，RPL 患者血清中的 HtrA4 水平低于对照组。这些发现表明 HtrA4 的耗竭可能与胎盘功能障碍有关。
