Gαi1蛋白促进短期低氧条件下表皮细胞迁移的实验研究

Gαi1 promotes migration of epidermal cells exposed to short-term hypoxia

目的观察低氧条件下表皮细胞迁移及运动变化,初步探究G蛋白偶联受体相关的G蛋白Gαi1在迁移调控中的作用。方法采用活细胞工作站观察比较HaCaT细胞在常氧及低氧处理24 h的迁移及运动情况,采用基因芯片技术筛选出早期关键调控基因,采用Western blot、免疫荧光以及慢病毒转染等方式,检测常氧及低氧条件下G蛋白表达变化,进一步观察细胞迁移情况。结果低氧条件下HaCaT细胞迁移较常氧条件迁移能力明显增强(P <0. 05); HaCaT细胞在低氧处理早期(前12 h)其运动速度及持久性明显强于常氧组。基因芯片、Western blot及免疫荧光检测结果显示,低氧条件下Gαi1蛋白呈现出高表达趋势(P <0. 05);干扰Gαi1蛋白后,HaCaT细胞迁移能力明显降低,常氧及低氧组差异无统计学意义;同时低氧早期运动能力及持久性也明显降低。结论低氧条件下表皮细胞迁移及运动能力明显增强。Gαi1蛋白在低氧促进表皮细胞迁移中起着极其重要的正向调控作用。

Objective To investigate the changes in the migration and motility of epidermal cells cultured under hypoxic conditions and the role of Gαi1 in promoting these changes.Methods A workstation for live cell imaging was used to observe the changes in the migration and motility of HaCaT cells cultured under hypoxic conditions for 24 h in comparison with cells in routine culture.Microarray chip technique was used to screen the differentially expressed genes that participated in the regulation of cell migration early after hypoxia exposure;Western blotting,immunofluorescence assay and lentivirus-mediated si RNA transfection were used to explore the role of Gαi1 protein in regulating the migration of HaCaT cells under hypoxic conditions.Results The migration ability of HaCaT cells was significantly enhanced under hypoxic conditions,and their movement rate and movement persistence were increased obviously early after hypoxic exposure( in the first 12 h) compared with the cells in normoxic culture.Western blotting and immunofluorescence assay showed that the exposure to hypoxia caused over-expression of Gαi1 protein in HaCaT cells.RNA interference of Gαi1 protein significantly lowered the migration ability of the cells irrespective of normoxic and hypoxic exposures and also resulted in obviously lowered cell motility and movement persistence.Conclusion Short-term hypoxic exposure significantly enhances the migration and movement of epidermal cells,and Gαi1 protein plays an important role in promoting the migration of epidermal cells under hypoxia.