[ALKBH5通过抑制上皮-间质转化抑制人滋养层细胞的迁移和侵袭]
[ALKBH5 suppresses migration and invasion of human trophoblast cells by inhibiting epithelial-mesenchymal transition].
作者信息
He Jianping, Li Xiaojuan, Lü Mengxin, Wang Jue, Tang Jian, Luo Shengjun, Qian Yuan
机构信息
Department of Medical Genetics and Prenatal Diagnosis, Kunming Maternal and Child Healthcare Hospital, Kunming 650031, China.
Department of Obstetrics, First Affiliated Hospital of Kunming Medical University, Kunming 650032, China.
出版信息
Nan Fang Yi Ke Da Xue Xue Bao. 2020 Dec 30;40(12):1720-1725. doi: 10.12122/j.issn.1673-4254.2020.12.04.
OBJECTIVE
To investigate the effects of ALKBH5 on migration, invasion and epithelial-mesenchymal transition (EMT) of human trophoblast cells.
METHODS
The expression plasmid of ALKBH5 or a negative control plasmid (ALKBH5-NC) was transfected in human trophoblast HTR-8 /SVneo cells, and the expressions of ALKBH5 mRNA and protein were detected by qRT-PCR and Western blotting. Transwell assay was used to assess the changes in migration and invasion abilities of the trophoblast cells after the transfection. Western blotting was performed to detect the expressions of EMT-related proteins in the cells including vimentin, fibronectin, E-cadherin, N-cadherin, MMP9 and MMP2.
RESULTS
ALKBH5 mRNA and protein expressions were significantly higher in ALKBH5 group than in the control group ( < 0.05). Over-expression of ALKBH5 significantly attenuated migration and invasion abilities of HTR-8/Svneo cells ( < 0.05). Compared with the control cells, the cells overexpressing ALKBH5 showed an up-regulated expression of E-cadherin and down-regulated expressions of vimentin, fibronectin, N-cadherin, MMP9 and MMP2 ( < 0.05).
CONCLUSIONS
ALKBH5 is involved in the pathogenesis of preeclampsia by inhibiting EMT of trophoblast cells and hence reducing their migration and invasion abilities.
目的
探讨ALKBH5对人滋养层细胞迁移、侵袭及上皮-间质转化（EMT）的影响。
方法
将ALKBH5表达质粒或阴性对照质粒（ALKBH5-NC）转染至人滋养层HTR-8/SVneo细胞中，采用qRT-PCR和蛋白质免疫印迹法检测ALKBH5 mRNA和蛋白的表达。采用Transwell实验评估转染后滋养层细胞迁移和侵袭能力的变化。通过蛋白质免疫印迹法检测细胞中波形蛋白、纤连蛋白、E-钙黏蛋白、N-钙黏蛋白、基质金属蛋白酶9（MMP9）和基质金属蛋白酶2（MMP2）等EMT相关蛋白的表达。
结果
ALKBH5组中ALKBH5 mRNA和蛋白表达显著高于对照组（P<0.05）。ALKBH5过表达显著减弱了HTR-8/Svneo细胞的迁移和侵袭能力（P<0.05）。与对照细胞相比，过表达ALKBH5的细胞E-钙黏蛋白表达上调，波形蛋白、纤连蛋白、N-钙黏蛋白、MMP9和MMP2表达下调（P<0.05）。
结论
ALKBH5通过抑制滋养层细胞的EMT，从而降低其迁移和侵袭能力，参与子痫前期的发病机制。
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