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以往研究证明,中性粒细胞明胶酶相关载脂蛋白NGAL(neutrophil gelatinase-associated lipocalin)与食管癌密切相关,改变NGAL表达能够对癌细胞的形态结构产生明显影响,但确切的作用机制不明.在酵母细胞中表达NGAL蛋白,柱层析分离纯化.筛选NGALR(NGAL receptor)高表达与弱表达的人食管癌细胞系EC1.71和EC109作为实验细胞模型.5-FAM标记NGAL蛋白,加入到细胞培养上清中,对比研究NGAL蛋白入胞情况、细胞形态学改变、细胞自噬体产生、自噬相关基因表达、细胞内铁离子与铁蛋白水平以及相关细胞信号转导激酶的活性等.结果表明,NGAL蛋白可经由内吞途径进入食管癌细胞发挥作用,致使细胞发生典型的自噬性形态结构变化,自噬体大量产生,自噬相关基因的表达发生相应变化,ERK被激活,但细胞内的铁离子与铁蛋白未受明显影响.上述结果提示,诱导细胞发生自噬是外源性NGAL蛋白经由内吞途径进入食管癌细胞发挥作用的机制之一,与NGAL蛋白跨膜转铁未见直接关联,而ERK信号转导途径可能参与了这一过程.
Previous studies have shown that neutrophil gelatinase-associated lipocalin NGAL (neutrophil gelatinase-associated lipocalin) and esophageal cancer are closely related to change the expression of NGAL can have a significant impact on the morphological structure of cancer cells, but the exact mechanism of action unknown. NGAL protein was expressed in yeast cells and purified by column chromatography.The human esophageal cancer cell lines EC1.71 and EC109 which were highly expressed and weakly expressed in NGALR (NGAL receptor) were screened as experimental cell models.5-FAM labeled NGAL protein was added into cells In the culture supernatants, we compared the influx of NGAL protein, cell morphological changes, autophagy production, autophagy-related gene expression, intracellular iron and ferritin levels, These results indicated that NGAL protein could enter the esophageal cancer cells via the endocytosis pathway, leading to typical autophagic morphological changes. The autophagosomes were produced in large quantities and the expression of autophagy-related genes changed accordingly. ERK was activated. However, Within the iron and ferritin were not significantly affected.The above results suggest that induction of cell autophagy is exogenous NGAL protein by endocytic One of the mechanisms of esophageal carcinoma cells play a role, NGAL protein transferrin no direct correlation, and ERK signaling pathway may be involved in this process.