Inhibition activity of a disulfide-stabilized diabody against basic fibroblast growth factor in lung cancer.
Inhibition activity of a disulfide-stabilized diabody against basic fibroblast growth factor in lung cancer.
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二硫键稳定的双抗体对肺癌碱性成纤维细胞生长因子的抑制活性
DOI：
10.18632/oncotarget.15556
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发表时间：
2017-03-21
期刊：
影响因子：
--
通讯作者：
Deng N
中科院分区：
文献类型：
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作者：
Cai Y;Yao S;Zhong J;Zhang J;Jiang H;Deng Y;Deng N
碱性成纤维细胞生长因子（bFGF）的过度表达在肺癌的发生、侵袭和转移中起着至关重要的作用。因此，抗碱性成纤维细胞生长因子的中和抗体可能会抑制肺癌的生长。本研究采用定点突变和重叠延伸PCR（SOE-PCR）技术，在单链抗体VH 44和VL 100位点构建了二硫键稳定的抗bFGF双抗体（ds-Diabody）。构建了双链抗体并在毕赤酵母中表达。我们发现抗bFGF双链抗体在体外能有效抑制人肺癌A549细胞的增殖、迁移和侵袭。在A549细胞中，抗bFGF的双链抗体可抑制bFGF诱导的下游信号调节因子磷酸化Akt和磷酸化MAPK的激活。在肺癌裸鼠移植瘤模型中，抗bFGF的双链抗体能显著抑制肿瘤生长，降低肿瘤组织中微血管和淋巴管密度。结果表明，抗bFGF双链抗体能够通过阻断bFGF信号通路和抑制肿瘤血管生成，有效抑制肺癌的生长，有望成为肺癌治疗的候选抗体药物。
The over-expression of basic fibroblast growth factor (bFGF) plays a crucial role in the development, invasion and metastasis of lung cancer. Therefore, neutralizing antibodies against bFGF may inhibit the growth of lung cancer. In this study, a Disulfide-stabilized diabody (ds-Diabody) against bFGF was constructed by site-directed mutation and overlap extension PCR (SOE-PCR) at the position of VH44 and VL100 in the scFv. The ds-Diabody was constructed and expressed in Pichia pastoris. We found that the ds-Diabody against bFGF could efficiently suppress the proliferation, migration and invasion of human lung cancer A549 cells in vitro. Moreover, in A549 cells, the ds-Diabody against bFGF could inhibit bFGF-induced activation of downstream signaling regulators, such as phospho-Akt and phospho-MAPK. In the nude mouse xenograft model of lung cancer, the ds-Diabody against bFGF could significantly inhibit tumor growth and decrease the densities of micro-vessels and lymphatic vessels in tumor tissue. Our data indicate that the ds-Diabody against bFGF could effectively suppress the lung cancer growth through blockade of bFGF signaling pathway and inhibition of tumor angiogenesis, which may make it a potential therapeutic candidate antibody drug for human lung cancer therapy.