RNA结合蛋白Aly/REF输出因子通过上调Yes相关蛋白表达促进胃癌细胞的增殖和迁移  

The RNA Binding Protein Aly/REF Export Factor Promotes the Proliferation and Migration of Gastric Cancer Cells via Up-regulating Yes-associated Protein

作　　者：李睿 邵爽 孟玮[2] 金丹[3] 郭纪伟 LI Rui;SHAO Shuang;MENG Wei;JIN Dan;GUO Ji-Wei(Gastroenterology Department,Binzhou Medical University Hospital,Binzhou 256603,Shandong,China;Laboratory Department,Binzhou Medical University Hospital,Binzhou 256603,Shandong,China;Medical Research Center,Binzhou Medical University Hospital,Binzhou 256603,Shandong,China)

机构地区：[1]滨州医学院附属医院消化内科,山东滨州256603 [2]滨州医学院附属医院检验科,山东滨州256603 [3]滨州医学院附属医院医学研究中心,山东滨州256603

出　　处：《中国生物化学与分子生物学报》2023年第4期622-630,共9页Chinese Journal of Biochemistry and Molecular Biology

基　　金：山东省高等学校“青创科技计划”(No.2020KJK002);“齐鲁卫生与健康杰出青年人才”项目支持。

摘　　要：胃癌是一种常见的发病率高、起病隐匿、侵袭性强的消化道恶性肿瘤,其发病机制错综复杂。RNA结合蛋白Aly/REF输出因子(RNAbinding protein Aly/REF export factor, ALYREF)作为一种RNA胞嘧啶的5-甲基化修饰(m~5C)后的结合蛋白质,在调控RNA出核及稳定其表达的过程中具有关键作用;Yes相关蛋白(yes-associated protein, YAP)作为Hippo-YAP信号通路中的关键分子,调控各种恶性肿瘤的生长和转移。本文旨在探讨ALYREF与YAP在促进胃癌细胞增殖和迁移中是否存在相关性,并初步探索这两者之间的作用机制。本文在胃癌细胞AGS和MKN-45中运用RT-PCR和Western印迹方法检测相关基因表达;CCK8试验检测细胞增殖;划痕试验和Trans-well试验分别检测细胞迁移和侵袭。结果显示,过表达ALYREF后YAP的表达增加(P<0.05),并且促进细胞的增殖、迁移和上皮细胞-间充质转化(epithelial-mesenchymal transition, EMT)(P<0.05);靶向敲低YAP,细胞的增殖、迁移和EMT能力显著抑制(P<0.05);过表达ALYREF且靶向敲除YAP与单独过表达ALYREF相比,YAP的表达显著降低(P<0.05),且细胞的增殖、迁移和EMT能力同样得到抑制(P<0.05);靶向敲低ALYREF,细胞的增殖、克隆形成和侵袭能力显著抑制(P<0.01)。综上所述,ALYREF通过上调YAP表达进而促进胃癌细胞的增殖和迁移。Gastric carcinoma(GC)is a common highly invasive cancer with a high disease rate and insidious onset,and its pathogenesis is complex.RNA binding protein Aly/REF export factor(ALYREF)has been reported as the(5-Methylcytosine)m 5C binding protein,which plays an important role in the nuclear export and stable expression of RNA.Yes-associated protein(YAP),the key molecule of the Hippo-YAP signaling pathway,regulates tumor growth and metastasis.Our research aims to investigate the correlation between ALYREF and YAP in cellular proliferation and migration of GC.The expressions of related genes were detected by RT-PCR and Western blot assays in the AGS and MKN-45 cells;the cellular viability and proliferation were detected by the CCK8 assay;the cell migration and invasion growth were detected by scratch and Trans-well assays.Our results showed that the mRNA and protein levels of YAP were increased(P<0.05)and cellular proliferation,migration growth and epithelial-mesenchymal transition(EMT)were enhanced(P<0.05)in the AGS and MKN-45 cells with transfection of ALYREF.However,the cellular proliferation and metastasis were significantly induced(P<0.05)in the AGS and MKN-45 cells with knockdown of YAP.Additionally,the mRNA and protein levels of YAP were recovered(P<0.05)whereby the cellular proliferation and migration were revived in the AGS and MKN-45 cells with co-transfection with ALYREF and shYAP(P<0.05),but were lower than the individual ALYREF group.Meanwhile,the cellular proliferation and metastasis were significantly induced(P<0.01)in the AGS and MKN-45 cells with knocking-down of ALYREF.In conclusion,the cellular proliferation and migration growth were promoted by ALYRERF via up-regulation of YAP in GC cells.

关 键 词：胃癌 增殖和迁移 RNA结合蛋白Aly/REF输出因子 Yes相关蛋白 

分 类 号：Q291[生物学—细胞生物学]