Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences/Key Laboratory of Biology and Genetic Resources of Torpical Crops, Ministry of Agriculture, Haikou 571101, Hainan, China.
Mol Biol Rep. 2013 Apr;40(4):3381-8. doi: 10.1007/s11033-012-2415-0. Epub 2012 Dec 28.
Hemangioma is a tumor that causes vascular endothelial cell hyperplasia, which commonly occur in newborns. Angiogenesis inhibitor targets the processes of angiogenesis, including the proliferation of vascular endothelial cells. A DNA sequence named Scl was designed, recombined into Pichia Pastoris, expressed by fermenting the engineered strain in a bioreactor, and purified the recombinant Scl by SP-sepharose fast flow. Scl can inhibit CAM angiogenesis. Only 1 μg of Scl significantly suppressed the growth of CAM blood vessel, similar to that of 25 μg of angiostatin. Scl showed a strong cytotoxicity on hemangioma cell (ATCC CRL No. 2587). After the drug acted for 24 h, the OD 570 measured value of the PBS control group averaged 1.873, whereas that of the Sc1 drug group was 0.692 (P < 0.01). Using the DeadEndTM Fluorometric TUNEL System, the detection results showed that 92 % of hemangioma cell apoptosis was observed in the Scl protein group, but only 1.3 % in the PBS control group (P < 0.01). After 2 weeks of treatment with the hemangioma model (cock's wattle) of the PBS group, 151 blood vessels with 100 views (40×) were obtained, whereas 250 in the PBS group (P < 0.01). During the two-week medication, the hemangioma model of the PBS group increased by 1.18 cm, whereas only 0.58 cm in the Scl drug group (P < 0.01).
血管瘤是一种引起血管内皮细胞增生的肿瘤,常见于新生儿。血管生成抑制剂靶向血管生成过程,包括血管内皮细胞的增殖。设计了一个名为 Scl 的 DNA 序列,将其重组到毕赤酵母中,通过在生物反应器中发酵工程菌株进行表达,并通过 SP-琼脂糖快速流动层析纯化重组 Scl。Scl 可以抑制鸡胚尿囊膜血管生成。仅 1μg 的 Scl 就能显著抑制鸡胚尿囊膜血管的生长,与 25μg 的血管抑素相似。Scl 对血管瘤细胞(ATCC CRL No.2587)表现出很强的细胞毒性。药物作用 24 小时后,PBS 对照组 OD570 测量值平均为 1.873,而 Sc1 药物组为 0.692(P<0.01)。使用 DeadEndTM 荧光原位末端标记(TUNEL)系统检测结果表明,Scl 蛋白组中观察到 92%的血管瘤细胞发生凋亡,而 PBS 对照组中仅为 1.3%(P<0.01)。在 PBS 组的血管瘤模型(鸡冠)治疗 2 周后,获得了 100 个视野(40×)的 151 个血管,而 PBS 组为 250 个(P<0.01)。在两周的用药期间,PBS 组的血管瘤模型增加了 1.18cm,而 Scl 药物组仅增加了 0.58cm(P<0.01)。