NHE1抑制剂EIPA在肝癌细胞中的作用及机制  

Role and mechanism of NHE1 inhibitor EIPA in hepatocellular carcino- ma cells

作　　者：金海 刘倩 李婧 赵思宇 朱加兴 JIN Hai;LIU Qian;LI Jing;ZHAO Shiyu;ZHU Jiaxing(Department of Gastroenterology,Digestive Disease Hospital,Affiliated Hospital of Zunyi Medical University,Zunyi 563003,China)

机构地区：[1]遵义医科大学附属医院消化病医院、遵义医科大学附属医院消化内科,贵州遵义563000

出　　处：《中国病理生理杂志》2025年第3期427-432,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.82460466);贵州省科技计划项目(黔科合基础-ZK[2024]一般323号)。

摘　　要：目的:研究Na^(+)/H^(+)交换体1(Na^(+)/H^(+)exchanger 1, NHE1)抑制剂乙基异丙基阿米洛利(ethylisopropylamiloride, EIPA)对肝细胞癌细胞增殖、迁移、侵袭、细胞周期和凋亡的影响,初步探讨其作用机制。方法:体外培养人肝癌Huh7细胞,用不同浓度的EIPA作用于细胞。RT-qPCR及Western blot法检测Huh7细胞NHE1的mRNA及蛋白表达;kFluor488-EdU法检测Huh7细胞增殖能力;Transwell法检测Huh7细胞迁移和侵袭能力;流式细胞术检测Huh7细胞周期和凋亡的变化;PTI高速离子成像系统检测Huh7细胞内pH的改变。结果:EIPA显著抑制NHE1的mRNA及蛋白表达(P<0.01);显著抑制Huh7细胞增殖、迁移和侵袭(P<0.05或P<0.01),促进Huh7细胞凋亡(P<0.01),使细胞发生明显的S期阻滞(P<0.01);显著降低Huh7细胞内pH(P<0.01)。结论:NHE1抑制剂EIPA通过抑制NHE1表达及功能调控细胞内pH,进而影响肝癌细胞的增殖、迁移、侵袭、细胞周期及凋亡。NHE1可以作为肝癌治疗的潜在靶点。AIM:To investigate the effects of ethylisopropylamiloride(EIPA),an inhibitor of Na+/H+exchanger 1(NHE1),on the proliferation,migration,invasion,cell cycle and apoptosis of hepatocellular carcinoma cells,and to explore its mechanisms.METHODS:The human hepatocellular carcinoma cell line Huh7 was cultured in vitro,and treated with different concentrations of EIPA.The mRNA and protein expression of NHE1 in Huh7 cells was assessed by RT-qPCR and Western blot.The proliferation of Huh7 cells was examined by kFluor488-EdU staining.The migration and invasion of Huh7 cells were assessed by Transwell assays.The cell cycle distribution and apoptosis of these cells were detected by flow cytometry.Moreover,the intracellular pH was measured via the PTI high-speed ion imaging system.RE-SULTS:Treatment with EIPA significantly suppressed the mRNA and protein expression of NHE1(P<0.01).It significantly inhibited the proliferation,migration and invasion of Huh7 cells(P<0.05 or P<0.01),promoted their apoptosis(P<0.01),and caused significant S-phase arrest(P<0.01).It also significantly reduced the intracellular pH of Huh7 cells.CONCLUSION:The NHE1 inhibitor EIPA regulates the intracellular pH by inhibiting the expression and function of NHE1,which consequently affects the proliferation,migration,invasion,cell cycle,and apoptosis of Huh7 cells.Therefore,NHE1 may be a potential target for the treatment of hepatocellular carcinoma.

关 键 词：Na^(+)/H^(+)交换体1 乙基异丙基阿米洛利 肝细胞癌 肿瘤酸性微环境 

分 类 号：R735.7[医药卫生—肿瘤] R363[医药卫生—临床医学] R977.7