西北大学许冰教授课题组在《Cell Death & Differentiation》发表了题为“LINC00941 promotes CRC metastasis through preventing SMAD4 protein degradation and activating the TGF-β/SMAD2/3 signaling pathway”的文章,长链非编码RNA(long non-coding RNA, lncRNA)是指长度大于200个核苷酸、无蛋白编码功能的RNA。近年来研究表明lncRNA LINC 00941在多种癌症迁移和侵袭中发挥了重要作用,而在结直肠癌中的表达及作用尚不清楚。通过TCGA数据库检索分析及组织芯片原位杂交检测LINC 00941在结直肠癌中的表达并分析与临床指标的相关性,利用慢病毒转染构建LINC 00941上调及下调细胞系,并通过Transwell及裸鼠肝肺转移实验检测LINC 00941对结直肠癌侵袭和迁移的作用,利用co-immunoprecipitation (Co-IP)、RNA immunoprecipitation (RIP)及质谱技术检测与LINC 00941潜在结合的蛋白并分析其促进结直肠癌转移的机制。研究发现LINC 00941在结直肠癌中表达较对应的癌旁组织显著升高,且与患者的淋巴结转移及不良预后密切相关;在结直肠癌细胞系LoVo中下调LINC 00941的表达后,细胞的EMT和迁移侵袭能力抑制,而在HCT-116细胞中过表达LINC 00941则促进细胞的EMT和迁移侵袭。通过RIP及质谱检测发现LINC 00941可与SMAD4蛋白结合,蛋白截短实验证实结合区域主要位于MH2结构域。功能研究发现,LINC 00941可竞争性抑制β-TrCP与SMAD4的结合进而抑制其泛素化降解,从而激活TGF-β/SMAD2/3 信号通路促进结直肠癌细胞的EMT和侵袭转移。综上所述,LINC 00941在转移性结直肠癌中表达升高,与β-TrCP蛋白竞争性结合SMAD4的MH2结构域进而抑制其泛素化降解,从而激活TGF-β/SMAD2/3 信号通路促进结直肠癌细胞的EMT和侵袭转移,LINC 00941可作为诊断和治疗转移性结直肠癌的潜在标志物。
本文利用利用四正柏超敏ECL显影液(UltraSignal ECL Reagent,4 A Biotech Co., Ltd, China)检测HCT-116与LoVo细胞中相应蛋白的含量,显影清晰,敏感度高。
文章链接:
https://www.nature.com/articles/s41418-020-0596-y
IF:10.717
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