背景:通过内体囊泡回收整合素对于癌细胞的迁移至关重要,这导致胰腺癌的转移和毁灭性的癌症相关的死亡。所以,需要开发新的诊断和治疗分子,靶向内体囊泡的再循环。
方法:公共数据库,包括TCGA,ICGC,分析GSE21501、GSE28735和GENT以获得诊断和治疗靶标。为了揭示分子靶标的生物学作用和潜在机制,进行了各种分子生物学实验。
结果:首先,我们确定了UNC13D在胰腺癌患者中的过度表达(n=824)及其在四个独立的胰腺癌队列中的预后意义和高风险比(HR)(TCGA,n=178,p=0.014,HR=3.629;ICGC,n=91,p=0.000,HR=4.362;GSE21501,n=102,p=0.002,HR=2.339;GSE28735,n=45,p=0.022,HR=2.681)。此外,其表达与胰腺癌的临床病理进展有关。进一步的生物学研究表明,UNC13D通过调节FAK磷酸化将再循环内体的胞吐作用与粘着灶性周转耦合来调节胰腺癌细胞的迁移。免疫沉淀和免疫细胞化学显示整合素再循环期间内体中RAB11-UNC13D-FAK轴的形成。我们观察到UNC13D直接与FAK的FERM结构域相互作用,并以钙依赖性方式调节FAK磷酸化。最后,我们发现UNC13D和FAK的共表达显示最差的存活率(TCGA,p=0.000;ICGC,p=0.036;GSE28735,p=0.006)。
结论:我们强调UNC13D,一个新的预后因素,通过RAB11-UNC13D-FAK轴将整联蛋白再循环与局灶性粘附周转耦合以促进胰腺癌细胞的迁移,从而促进胰腺癌的进展。
BACKGROUND: Recycling of integrin via endosomal vesicles is critical for the migration of cancer cells, which leads to the metastasis of pancreatic cancer and devastating cancer-related death. So, new diagnostic and therapeutic molecules which target the recycling of endosomal vesicles need to be developed.
METHODS: Public databases including TCGA, ICGC, GSE21501, GSE28735, and GENT are analyzed to derive diagnostic and therapeutic targets. To reveal biological roles and underlying mechanisms of molecular targets, various molecular biological experiments were conducted.
RESULTS: First, we identified UNC13D\'s overexpression in patients with pancreatic cancer (n = 824) and its prognostic significance and high hazard ratio (HR) in four independent pancreatic cancer cohorts (TCGA, n = 178, p = 0.014, HR = 3.629; ICGC, n = 91, p = 0.000, HR = 4.362; GSE21501, n = 102, p = 0.002, HR = 2.339; GSE28735, n = 45, p = 0.022, HR = 2.681). Additionally, its expression is associated with the clinicopathological progression of pancreatic cancer. Further biological studies have shown that UNC13D regulates the migration of pancreatic cancer cells by coupling the exocytosis of recycling endosomes with focal adhesion turnover via the regulation of FAK phosphorylation. Immunoprecipitation and immunocytochemistry showed the formation of the RAB11-UNC13D-FAK axis in endosomes during integrin recycling. We observed that UNC13D directly interacted with the FERM domain of FAK and regulated FAK phosphorylation in a calcium-dependent manner. Finally, we found co-expression of UNC13D and FAK showed the poorest survival (TCGA, p = 0.000; ICGC, p = 0.036; GSE28735, p = 0.006).
CONCLUSIONS: We highlight that UNC13D, a novel prognostic factor, promotes pancreatic cancer progression by coupling integrin recycling with focal adhesion turnover via the RAB11-UNC13D-FAK axis for the migration of pancreatic cancer cells.