长链非编码RNA AC087388.1靶向PABPC1对食管鳞癌作用机制  

Mechanism of action of long non-coding RNA AC087388.1 targeting PABPC1 on esophageal squamous cell carcinoma

作　　者：仲瀚 戈艳蕾 甘俊清 金叶 郑璇 刘子情 孙国贵 ZHONG Han;GE Yan-lei;GAN Jun-qing;JIN Ye;ZHENG Xuan;LIU Zi-qing;SUN Guo-gui(North China University of Science and Technology Affiliated Hospital;Dept of Hebei Key Laboratory of Medical-Industrial Intergration Precision Medicine;Dept of Tangshan Key Laboratory of Medical-Industrial Intergration Precision Medicine;Clinical Medicine School,North China University of Science and Technology,Tangshan Hebei 063000,China;School of Public Health,North China University of Science and Technology;School of Pharmacy,North China University of Science and Technology,Tangshan Hebei 063210,China)

机构地区：[1]华北理工大学附属医院 [2]河北省医工融合精准医疗重点实验室 [3]唐山市医工融合精准医疗重点实验室 [4]华北理工大学临床医学院,河北唐山063000 [5]华北理工大学公共卫生学院 [6]华北理工大学药学院,河北唐山063210

出　　处：《中国药理学通报》2025年第5期926-935,共10页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金面上项目(No 82472636);河北省创新能力提升计划项目(No 235A2403D);河北省教育厅河北实验教学及教学实验室建设项目(No 81);华北理工大学公共卫生学院高水平科研创新团队建设计划(No KYTD202309)。

摘　　要：目的探讨长链非编码RNA(long non-coding RNA,lncRNA)AC087388.1和细胞质多聚腺苷酸结合蛋白1(poly(A)binding protein cytoplasmic 1,PABPC1)在食管鳞癌(esophageal squamous cell carcinoma,ESCC)中的作用与机制。方法采用实时荧光定量聚合酶链反应(real-time quantitative polymerase chain reaction,RT-qPCR)和荧光原位杂交实验检测AC087388.1在ESCC组织和细胞中的表达水平并分析其临床相关性。采用细胞计数试剂盒-8(cell counting kit-8,CCK-8)、克隆形成、划痕和Transwell侵袭实验分别检测敲降AC087388.1对ESCC细胞的活性、增殖、迁移和侵袭能力的影响,以及在细胞中的亚定位。RNA Pull Down和蛋白免疫印迹(Western blot)实验验证在ESCC细胞中AC087388.1和PABPC1互相作用。进行挽救实验检测AC087388.1靶向PABPC1对ESCC细胞的影响。结果AC087388.1在ESCC组织和细胞中高表达,且与ESCC患者临床分期正相关,主要定位于细胞质中。敲降AC087388.1可抑制ESCC细胞活性、增殖、迁移和侵袭能力。在RNA Pull Down-MS实验的结果中选择PABPC1进行后续实验,通过RNA Pull Down和Western blot实验验证了AC087388.1与PABPC1相结合。挽救实验验证过表达AC087388.1可以逆转敲降PABPC1对ESCC细胞活性、增殖、迁移和侵袭的影响。结论AC087388.1的高表达与临床分期相关,可能是ESCC进展的危险因素,其通过靶向PABPC1促进ESCC细胞增殖、迁移和侵袭,其可能成为ESCC的诊断和治疗的一个新的生物标志物。Aim To explore the roles and mechanisms of long non-coding RNA(lncRNA)AC087388.1 and poly(A)binding protein cytoplasmic 1(PABPC1)in esophageal squamous cell carcinoma(ESCC).Methods The expression level of AC087388.1 in ESCC tissues and cells was detected by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)and fluorescence in situ hybridization experiments and its clinical relevance was analyzed.Cell counting kit-8(CCK-8),clone formation,scratch and Transwell invasion assays were used to detect the effects of knockdown of AC087388.1 on the cell viability,proliferation,migratory,and invasion of ESCC cells respectively ability,and sub-localization in cells.RNA pull down and Western blot experiments were employed to verify the interaction between AC087388.1 and PABPC1 in ESCC cells.Salvage experiments were performed to detect the effect of AC087388.1 targeting PABPC1 on ESCC cells.Results AC087388.1 was highly expressed in ESCC tissues and cells and positively correlated with clinical stage of ESCC patients,mainly localized in cytoplasm.Knockdown AC087388.1 inhibited ESCC cell viability,proliferation,migration and invasionability.PABPC1 was selected from the results of RNA Pull Down-MS experiments for subsequent experiments,and AC087388.1 was verified to bind to PABPC1 by RNA Pull Down and Western blot experiments.Overexpression of AC087388.1 was verified by rescue experiment to reverse the effects of knockdown of PABPC1 on ESCC cell viability,proliferation,migration and invasion.Conclusions High expression of AC087388.1 correlates with clinical stage and may be a risk factor for ESCC progression.AC087388.1 promotes the cell viability,proliferation,migration and invasive ability of ESCC cells by targeting PABPC1,which may be a novel biomarker for the diagnosis and treatment of ESCC.

关 键 词：食管鳞状细胞癌 lnc-AC087388.1 细胞质多聚腺苷酸结合蛋白1 增殖 迁移 侵袭 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学] R341[医药卫生—基础医学] R342.2R730.261R735.1