缺氧下调FTO并促进小鼠肺动脉平滑肌细胞迁移

Hypoxia down⁃regulates FTO and promotes migration of mouse pulmonary artery smooth muscle cells

目的:探究脂肪质量与肥胖相关蛋白(fat mass and obesity associated protein,FTO)对缺氧小鼠肺动脉平滑肌细胞(mouse pulmonary artery smooth muscle cell,MPASMC)迁移能力的影响。方法:体外培养MPASMC,用三气培养箱构建缺氧MPASMC模型;免疫印迹实验用于检测FTO蛋白表达;FTO过表达以及敲低质粒转染MPASMC以过表达或抑制FTO表达;细胞划痕实验检测细胞迁移能力。结果:缺氧培养后的MPASMC中FTO蛋白表达明显下调,划痕实验提示细胞迁移加快,并在过表达FTO后明显减慢;常氧状态下敲低FTO表达出现了类似缺氧培养的“促迁移”表型。结论:在体外条件下MPASMC缺氧培养后FTO蛋白水平下调,细胞迁移能力增强,并且FTO可能是促进MPASMC迁移的关键调控基因。

Objective:The current study aims to investigate the effect of fat mass and obesity associated protein(FTO)on the migration of hypoxia⁃inoluced pulmonary artery smooth muscle cells(PASMC).Methods:MPASMC was cultured in vitro.The hypoxia model of MPASMC was established in a three⁃gas incubator(37℃,1%O2,5%CO2).The expression of FTO protein was detected by Western blot.FTO overexpression and knockdown plasmids were transfected into MPASMC to over⁃express or inhibit FTO expression,respectively.Cell wound healing assay was used to detect cell migration ability.Results:The expression of FTO protein in MPASMC after hypoxia culture was significantly down⁃regulated.Wound healing assay indicated that MPASMC migration was accelerated after hypoxia culture and was significantly slowed down after FTO overexpression in MPASMC.Knockdown FTO expression in MPASMC in normoxia showed a“pro⁃migration”phenotype similar to hypoxic culture.Conclusion:Under hypoxic conditions,the expression of FTO protein is down⁃regulated in MPASMC and the cells migration ability is enhanced.FTO may be a key regulator of MPASMC migration.