The L‐fucose‐specific lectin from Aspergillus niger (ANL), isolated from the corneal smears of a keratitis patient was reported earlier. Here, we examined the interaction of ANL with human hepatocellular and colon cancer cells, evaluated its anti‐cancer activity and diagnostic potential to detect aberrantly glycosylated tumour‐associated serum glycoproteins such as alpha‐fetoprotein (AFP). We observed that ANL strongly bound to both HepG2 and HT‐29 cell‐lines and this interaction was effectively blocked with L‐fucose and mucin in a dose and time‐dependent manner with an IC₅₀ of 1.25 and 5 μg/mL for HepG2 and HT‐29 cells respectively at 48 hours. ANL treatment increased hypodiploidy and decreased the number of HepG2 cell in G₀‐G₁ phase at both 24 and 48 hours. Furthermore, ANL increased the level of apoptosis in both HepG2 and HT‐29 cells in a time‐dependent manner via enhanced production of reactive oxygen species and altered mitochondrial membrane potential, indicative of intrinsic apoptotis pathway activation. Immunoblot analysis confirmed the time‐dependent elevation of levels of cytochrome c, initiator caspase‐9 and activation of caspase‐3. ANL immunohistochemistry on colon cancer tissue and quantification of AFP in HCC patient serum samples by developing an ANL‐anti‐AFP antibody sandwich enzyme‐linked immunosorbent assay confirmed the diagnostic potential of ANL. Here, interaction of ANL with AFP could be effectively blocked in the presence of competing fucose‐bearing glycans. We found ANL to be more sensitive than Lens culinaris lectin, a well‐known fucose‐specific lectin and currently used diagnostic agent. ANL can be further explored as a diagnostic and anti‐cancer agent.

中文翻译：

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黑曲霉的岩藻糖特异性凝集素ANL通过诱导肝细胞和结肠癌细胞的内在凋亡途径而具有抗癌活性

较早前报道了从黑曲霉（ANL）分离出的L-岩藻糖特异性凝集素，该凝集素从一名角膜炎患者的角膜涂片中分离出来。在这里，我们检查了ANL与人类肝细胞和结肠癌细胞的相互作用，评估了其抗癌活性和诊断潜力，以检测异常糖基化的肿瘤相关血清糖蛋白，例如甲胎蛋白（AFP）。我们观察到ANL与HepG2和HT-29细胞系都牢固结合，并且这种相互作用被L-岩藻糖和粘蛋白以剂量和时间依赖的方式有效阻断，HepG2和IC的IC ₅₀为1.25和5μg/ mL。 HT-29细胞分别在48小时内。ANL处理增加了二倍体，减少了G ₀ -G ₁中HepG2细胞的数量分别在24小时和48小时内进入阶段。此外，ANL通过增加活性氧的产生和改变线粒体膜电位，从而以时间依赖性的方式增加了HepG2和HT-29细胞的凋亡水平，这表明内在的凋亡途径活化。免疫印迹分析证实了细胞色素c，启动子caspase-9和caspase-3激活水平随时间的升高。通过开发ANL‐anti-AFP抗体夹心酶联免疫吸附测定法对结肠癌组织进行ANL免疫组织化学和HCC患者血清样本中AFP的定量测定，证实了ANL的诊断潜力。在这种情况下，当存在带有岩藻糖的竞争聚糖时，可以有效地阻止ANL与AFP的相互作用。我们发现ANL比lens culinaris更敏感血凝素是一种著名的岩藻糖特异性凝集素，目前是诊断试剂。可以进一步探索ANL作为诊断和抗癌药。