Jianping He1, Xiaojuan Li2,3, Mengxin Lü1, Jue Wang4, Jian Tang1, Shengjun Luo1, Yuan Qian1,2,4,5,6,7. 1. Department of Medical Genetics and Prenatal Diagnosis, Kunming Maternal and Child Healthcare Hospital, Kunming 650031, China. 2. Department of Obstetrics, First Affiliated Hospital of Kunming Medical University, Kunming 650032, China. 3. Department of Obstetrics, First People's Hospital of Yunnan Province, Kunming 650032, China. 4. Prenatal Diagnosis Room, Clinical Laboratory, First Affiliated Hospital of Kunming Medical University, Kunming 650032, China. 5. Key Laboratory of Laboratory Medicine of Yunnan Province, Kunming 650032, China. 6. Institute of Practical Diagnosis of Yunnan Province, Kunming 650032, China; 7Research Institute in Yunnan Province, Kunming 650032, China. 7. Research Institute in Yunnan Province, Kunming 650032, China.
Abstract
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 (P < 0.05). Over-expression of ALKBH5 significantly attenuated migration and invasion abilities of HTR-8/Svneo cells (P < 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 (P < 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.
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 (P < 0.05). Over-expression of ALKBH5 significantly attenuated migration and invasion abilities of HTR-8/Svneo cells (P < 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 (P < 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.
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