首页|PIEZO1影响HaCaT细胞的趋电性迁移

PIEZO1影响HaCaT细胞的趋电性迁移

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目的 伤口中心产生的内源性电场是指导细胞定向迁移促进伤口愈合的重要因子.PIEZO1是机械门控阳离子通道家族成员之一,它参与细胞迁移,影响细胞的趋化迁移,并且受到电压的调节,在伤口愈合过程中发挥重要作用.但PIEZO1是否参与影响电场指导的细胞定向迁移过程尚不清晰,本文以HaCaT细胞为模型探讨PIEZO 1及其下游相关蛋白质对细胞趋电性迁移的影响.方法 应用活细胞工作站追踪HaCaT细胞在微直流电场中的迁移,使用抑制剂及RNAi技术调控PIEZO 1功能和表达,研究PIEZO1对于细胞趋电性迁移的影响;用蛋白质印迹(Western blot)检验细胞的整合素(integrin)β1与FAK磷酸化水平在电场作用下的变化情况,探讨PIEZO1与integrin β1及FAK等对电场信号的响应及细胞趋电性迁移的影响.结果 PIEZO1 广谱抑制剂钌红、GsMTx4和RNAi处理显著抑制了 HaCaT细胞向正极趋电性迁移的能力;电场和GsMTx4单独作用升高FAK磷酸化和integrin β1表达,GsMTx4阻止电场进一步升高FAK的磷酸化水平和integrin β1表达;siRNA干扰PIEZO 1表达后显著下调FAK的磷酸化水平,并且电场对FAK磷酸化和integrin β1表达的促进作用受到抑制;Integrin β1和FAK的抑制剂使HaCaT细胞的趋电性迁移能力均显著降低.结论 P1EZO1参与HaCaT细胞的趋电性迁移,是影响HaCaT细胞趋电性迁移的重要分子之一;抑制integrin β1和FAK会影响HaCaT细胞的电性迁移;电场信号可能通过PIEZO 1介导的信号通路调控integrin β1的表达和FAK的活化进而影响细胞趋电性迁移.
PIEZO1 Channel is Involved in Electric Field Guided Cell Migration
Objective Disruption of epithelial layer may instantaneously induce the generation of endogenous electric fields,which was proved to play an important role in guiding the cell migration and promoting wound healing.PIEZO 1 is a kind of mechanic sensitive channel,may be regulated by voltage,is proved to involve in chemotactic migration of cells and play an important role in the process of wound healing.In this paper,the role of PIEZO 1 and its downstream proteins FAK and integrin β1 in the electric field guided cell migration were investigated by HaCaT cells(human immortalized keratinocyte).Methods Cell migration was tracked by Living Cell Imaging System in directed current(DC)electric field(EF).Inhibitors and RNAi techniques were applied to study the function of PIEZO 1 and other related proteins in electric fields.Western blot was used to detect the expression and phosphorylation levels of integrin β1 and FAK in electric field guided migration under EF stimulation.Results Piezo1 RNAi as well as Ruthenium red and GsMTx4 treatment all significantly inhibited the electrotaxis of HaCaT cells.Electric field stimulation with GsMTx4 treatment alone increased FAK phosphorylation level and the expression of integrin β1.Electric field promoted the expression level of integrin β1 and the phosphorylation level of FAK.Inhibiting the expression of PIEZO 1 by RNAi significantly attenuated the phosphorylation level of FAK under EF stimulation.Inhibition of integrin β1 and FAK by inhibitor significantly decrease the electric field guided cell migration.Conclusion PIEZO1 as well as integrin β1 and FAK are involved in the electric field guided cell migration of HaCaT cells.Electric field signals regulate the expression of integrin β1 and the activation of FAK through PIEZO 1-mediated signal pathway to orchestrate cell migration.

electric fieldselectrotaxiscell migrationHaCaTPIEZO1integrin β1FAK

张桂诚、龚鹏、王一凡、赵三军

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云南师范大学生命科学学院,昆明 650500

电场 趋电性 细胞迁移 HaCaT细胞 PIEZO1 整合素β1 FAK

国家自然科学基金国家自然科学基金

8206035581660519

2024

生物化学与生物物理进展
中国科学院生物物理研究所,中国生物物理学会

生物化学与生物物理进展

CSTPCD北大核心
影响因子:0.476
ISSN:1000-3282
年,卷(期):2024.51(3)
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