According to their sugar recognition specificity, plant lectins are proposed as bioactive proteins with potential in cancer treatment and diagnosis. Helja is a mannose-specific jacalin-like lectin from sunflower which was shown to inhibit the growth of certain fungi. Here, we report its recombinant expression in a prokaryotic system and its activity in neurobalstoma cells. Helja coding sequence was fused to the pET-32 EK/LIC, the enterokinase/Ligation-independent cloning vector and a 35 kDa protein was obtained in Escherichia coli representing Helja coupled to thioredoxin (Trx). The identity of this protein was verified using anti-Helja antibodies. This chimera, named Trx-rHelja, was enriched in the soluble bacterial extracts and was purified using Ni+2-Sepharose and d-mannose-agarose chromatography. Trx-rHelja and the enterokinase-released recombinant Helja (rHelja) both displayed toxicity on human SH-SY5Y neuroblastomas. rHelja decreased the viability of these tumor cells by 75% according to the tetrazolium reduction assay, and microscopic analyses revealed that the cell morphology was disturbed. Thus, the stellate cells of the monolayer became spheroids and were isolated. Our results indicate that rHelja is a promising tool for the development of diagnostic or therapeutic methods for neuroblastoma cells, the most common solid tumors in childhood.

译文

:根据其糖识别特异性,提出了植物凝集素作为具有癌症治疗和诊断潜力的生物活性蛋白。 Helja是一种来自向日葵的甘露糖特异性贾卡林样凝集素,被证明可以抑制某些真菌的生长。在这里,我们报告其在原核系统中的重组表达及其在神经原发瘤细胞中的活性。将Helja编码序列与pET-32 EK / LIC融合,肠激酶/连接独立的克隆载体,并在大肠杆菌中获得35 kDa蛋白,代表Helja与硫氧还蛋白(Trx)偶联。使用抗Helja抗体验证了该蛋白质的身份。该嵌合体名为Trx-rHelja,富含可溶细菌提取物,并使用Ni 2-Sepharose和d-甘露糖-琼脂糖层析纯化。 Trx-rHelja和肠激酶释放的重组Helja(rHelja)均对人SH-SY5Y神经母细胞瘤具有毒性。根据四唑鎓还原法,rHelja将这些肿瘤细胞的活力降低了75%,显微镜分析表明细胞形态受到干扰。因此,单层的星状细胞变成球状并被分离。我们的结果表明,rHelja是开发成神经细胞瘤细胞(儿童时期最常见的实体瘤)的诊断或治疗方法的有前途的工具。

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