目的:探讨着丝粒蛋白F(centromere protein F,CENPF)在腺样囊性癌(adenoid cystic carcinoma,ACC)发生发展中的作用和分子机制.方法:通过小干扰RNA转染敲低ACC细胞中的CENPF.通过细胞计数试剂盒(cell counting kit-8,CCK-8)、划痕实验和Transwell测定评估敲低CENPF对ACC细胞增殖、迁移、侵袭的影响;通过蛋白免疫印迹实验分析改变CENPF表达后ACC细胞中磷脂酰肌醇-3-激酶(phosphatidylinositol-3-kinase,PI3K)/丝氨酸/苏氨酸蛋白激酶(serine/threonine protein kinase,AKT)/哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)信号通路相关蛋白表达变化,通过CCK-8、划痕实验和Transwell测定评估敲低CENPF联合PI3K抑制剂BKM-120对ACC细胞增殖、迁移、侵袭的影响.结果:敲低ACC-M中CENPF表达后,CCK-8实验证明其增殖能力减弱;划痕实验表明其迁移能力减弱,Transwell实验表明其侵袭能力减弱,PI3K/AKT通路关键蛋白p-PI3K、p-AKT和p-mTOR蛋白表达水平下降.在下调CENPF的ACC-M中加入PI3K抑制剂,PI3K/AKT通路关键蛋白表达水平进一步下降.此外,加入PI3K抑制剂后,CENPF下调的ACC-M增殖、迁移、侵袭能力较单纯下调CENPF的ACC-M进一步减弱.结论:CENPF通过调控PI3K/AKT信号通路参与ACC进展.
Impact of CENPF on Progression of Adenoid Cystic Carcinoma through Regulation of PI3K/AKT Signaling Pathway
Objective:To investigate the role and molecular mechanisms of CENPF in the pathogenesis and devel-opment of adenoid cystic carcinoma(ACC).Methods:CENPF in ACC cells was knocked down by transfection with small interfering RNA(siRNA).The effects of CENPF knockdown on the proliferation,migration,and invasion of ACC cells were evaluated using CCK-8,scratch assay,and transwell assay,respectively.Changes in the activity of phosphatidylinositol-3-kinase/serine/threonine protein kinase/mammalian target of rapamycin(PI3K/AKT/mTOR)signaling pathway-related proteins in ACC cells after altering CENPF expression were analyzed by Western blot.Finally,the combined effect of CENPF knockdown and PI3K inhibitor BKM-120 on the proliferation,migra-tion,and invasion of ACC cells was assessed using CCK-8,scratch assay,and transwell assay.Results:Knockdown of CENPF expression in ACC-M cells resulted in decreased proliferation,migration,and invasion abilities as demon-strated by CCK-8,scratch assay,and transwell assay,respectively.The expression levels of key proteins in the PI3K/AKT pathway,including p-PI3K,p-AKT,and p-mTOR,were reduced.Furthermore,the expression levels of these proteins further decreased when PI3K inhibitor was added to CENPF-knockdown ACC-M cells.Additional-ly,the proliferation,migration,and invasion abilities of ACC-M cells with CENPF knockdown were further weak-ened when PI3K inhibitor was added compared to CENPF knockdown alone.Conclusion:CENPF regulates the pro-gression of ACC by modulating the PI3K/AKT signaling pathway.
centromere protein Fadenoid cystic carci-nomaPI3K/AKT signaling pathwaymechanistic study
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