Crumbs3 is a critical factor that regulates invasion and metastasis of colon adenocarcinoma via the specific interaction with FGFR1.
Crumbs3 is a critical factor that regulates invasion and metastasis of colon adenocarcinoma via the specific interaction with FGFR1.
复制标题
Crumbs3是通过与FGFR1特异性相互作用调节结肠腺癌侵袭和转移的关键因子。
作者：
Iioka H;Saito K;Sakaguchi M;Tachibana T;Homma K;Kondo E.
上皮细胞极性调节因子Crumbs 3（Crb 3）是果蝇Crb基因家族中的哺乳动物同源物，最初被认为是胚胎发育的重要因子。它最近被牵连在肿瘤抑制，虽然其具体功能是有争议的。我们发现Crb 3对人结肠腺癌细胞的侵袭和转移有明显的促进作用。Crb 3对肿瘤迁移的中心性由患者组织的结肠腺癌的浸润前沿和转移灶的强表达支持。因此，通过CRISPR-Cas9系统产生了来自人结肠腺癌的两种不同的Crb 3敲除（KO）细胞系Crb 3-KO（Crb 3-/-）DLD-1和Crb 3-KO WiDr。与野生型DLD-1相比，Crb 3-KOLD-1细胞表现出体外细胞迁移率的丧失和体内肝转移酶的显著抑制。与DLD-1不同，Crb 3-KOWiDr的移动性和转移性不受影响，这与野生型WiDr相似。Crb 3免疫共沉淀蛋白的蛋白质组分析鉴定了不同的成纤维细胞生长因子受体（FGFR）同种型，其通过其细胞内结构域特异性结合Crb 3同种型a。在DLD-1中，Crb 3显示出FGFR 1的膜定位，导致其功能活化，而在没有FGFR 1表达的WiDr中，Crb 3与细胞质FGFR 4结合，导致细胞生长。Crb 3和FGFR 1在原发性和转移性结直肠癌组织中的相关表达一致。综上所述，Crb 3通过与结肠癌细胞上的FGFR 1特异性相互作用，显著加速细胞迁移，即人类结肠癌的侵袭和转移。
Epithelial cell polarity regulatorCrumbs3(Crb3), a mammalian homolog within theDrosophila Crbgene family, was initially identified as an essential embryonic development factor. It is recently implicated in tumor suppression, though its specific functions are controversial. We here demonstrate thatCrb3strongly promotes tumor invasion and metastasis of human colon adenocarcinoma cells. Crb3 centrality to tumor migration was supported by strong expression at invasive front and metastatic foci of colonic adenocarcinoma of the patient tissues. Accordingly, two differentCrb3‐knockout (KO) lines,Crb3‐KO (Crb3−/−) DLD‐1 andCrb3‐KO WiDr from human colonic adenocarcinomas, were generated by the CRISPR‐Cas9 system.Crb3‐KODLD‐1 cells exhibited loss of cellular mobilityin vitroand dramatic suppression of liver metastasesin vivoin contrast to the wild type of DLD‐1. Unlike DLD‐1,Crb3‐KOWiDr mobility and metastasis were unaffected, which were similar to wild‐type WiDr. Proteome analysis of Crb3‐coimmunopreciptated proteins identified different respective fibroblast growth factor receptor (FGFR) isotypes specifically bound to Crb3 isoform a through their intracellular domain. In DLD‐1, Crb3 showed membranous localization of FGFR1 leading to its functional activation, whereas Crb3 bound to cytoplasmic FGFR4 in WiDr without FGFR1 expression, leading to cellular growth. Correlative expression between Crb3 and FGFR1 was consistently detected in primary and metastatic colorectal cancer patient tissues. Taking these together, Crb3 critically accelerates cell migration, namely invasion and metastasis of human colon cancers, through specific interaction to FGFR1 on colon cancer cells.