State Key Laboratory of Silkworm Genome Biology, College of Biotechnology, Southwest University, Chongqing, 400715, China.
Transgenic Res. 2019 Dec;28(5-6):601-609. doi: 10.1007/s11248-019-00173-y. Epub 2019 Sep 20.
Recombinant human vascular endothelial growth factor (rhVEGF) has important applications in therapeutic angiogenesis and inhibition of VEGF-mediated pathological angiogenesis. Previous studies have shown that rhVEGF can be produced in several expression systems, including Escherichia coli, yeasts, insect cells and mammalian cells. However, little is known regarding the effective production of this protein in organs of live organisms. Here, we report for the first time the expression and characterization of rhVEGF165 in the middle silk gland (MSG) of the transgenic silkworm line S1-V165. Our results confirmed that (1) rhVEGF165 was highly expressed in MSG cells and was secreted into the cocoon of S1-V165; (2) the dimeric form of rhVEGF165 could be easily dissolved from S1-V165 cocoons using an alkaline solution; (3) rhVEGF165 extracted from S1-V165 cocoons exhibited slightly better cell proliferative activity than the hVEGF165 standard in cultured human umbilical vein endothelial cells. This study provides an alternative strategy for the production of bioactive rhVEGF165 using the MSG of transgenic silkworms.
重组人血管内皮生长因子（rhVEGF）在治疗性血管生成和抑制 VEGF 介导的病理性血管生成方面有重要应用。先前的研究表明 rhVEGF 可以在多种表达系统中产生，包括大肠杆菌、酵母、昆虫细胞和哺乳动物细胞。然而，对于这种蛋白质在活体生物器官中的有效生产知之甚少。在这里，我们首次报道了 rhVEGF165 在转基因家蚕 S1-V165 的中肠（MSG）中的表达和特性。我们的结果证实：（1）rhVEGF165 在 MSG 细胞中高度表达，并分泌到 S1-V165 的茧中；（2）rhVEGF165 的二聚体形式可以用碱性溶液从 S1-V165 茧中轻易溶解；（3）从 S1-V165 茧中提取的 rhVEGF165 在培养的人脐静脉内皮细胞中表现出比 hVEGF165 标准品稍好的细胞增殖活性。本研究为利用转基因家蚕的 MSG 生产生物活性 rhVEGF165 提供了一种替代策略。
