抗骨桥蛋白单链抗体可抑制乳腺癌转移和血管生成
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摘要
骨桥蛋白(Osteopontin，OPN)与肿瘤的生长和转移有密切的关系。为系统研究抗OPN抗体在抑制肿瘤转移过程中的作用，探讨OPN作为肿瘤转移治疗靶点的可能性，本实验在建立噬菌体抗体展示平台的基础上，筛选得到抗OPN单克隆单链抗体，构建并表达获得抗OPN单链抗体-Fc融合蛋白；研究抗OPN单链抗体-Fe融合蛋白在体内外对乳腺癌转移和血管生成的作用。结果表明抗OPN单链抗体-Fc融合蛋白1A12和2H8不但能够抑制乳腺癌细胞MDA-MB-435s在体外的粘附、侵袭、迁移和锚定非依赖生长，而且可以抑制OPN体内外的促血管生成作用；该血管生成抑制作用是通过抑制NF-κB介导的细胞生存调节通路，促进内皮细胞凋亡而实现的。在体内模型中，1A12和2H8能够抑制MDA-MB-435s在裸鼠体内的原发肿瘤生长和自发性肺转移，并能够降低肿瘤组织中微血管密度，增加肿瘤内皮细胞凋亡。本研究发现OPN是抑制肿瘤转移的有效治疗靶点，抗OPN单链抗体-Fc融合蛋白可以通过抑制肿瘤细胞的转移活性同时阻碍肿瘤血管生成，从而达到抑制肿瘤转移的目的。
Osteopontin (OPN) has been associated with tumor growth and metastasis. In order to study the function of anti-OPN antibodies in inhibiting tumor metastasis, we have raised several anti-OPN single-chain Fv (scFv) antibodies from phage antibody library and expressed them as scFv-Fc fusion proteins. We studied their anti-tumor effects in vitro and in vivo. Two anti-OPN scFv-Fc fusion proteins 1A12 and 2H8 were able to inhibit MDA-MB-435s breast cancer cell attachment, invasion, migration and colony formation in soft agar. 1A12 and 2H8 showed markedly inhibitory effects toward angiogenesis, and this inhibitory effect was achieved by inhibiting NF-kB mediated cell survival pathway and increasing endothelial cell apoptosis. In vivo studies showed 1A12 and 2H8 delayed primary tumor growth and reduced spontaneous lung metastasis in breast cancer xenograft model; furthermore, they reduced tumor microvessel density and increased tumor endothelial cell apoptosis. The present study has demonstrated OPN is an effective target in the treatment of tumor, and anti-OPN scFv-Fc fusion proteins could inhibit tumor metastasis by inhibiting angiogenesis and breast cancer cell migration.
Osteopontin (OPN) has been associated with tumor growth and metastasis. In order to study the function of anti-OPN antibodies in inhibiting tumor metastasis, we have raised several anti-OPN single-chain Fv (scFv) antibodies from phage antibody library and expressed them as scFv-Fc fusion proteins. We studied their anti-tumor effects in vitro and in vivo. Two anti-OPN scFv-Fc fusion proteins 1A12 and 2H8 were able to inhibit MDA-MB-435s breast cancer cell attachment, invasion, migration and colony formation in soft agar. 1A12 and 2H8 showed markedly inhibitory effects toward angiogenesis, and this inhibitory effect was achieved by inhibiting NF-kB mediated cell survival pathway and increasing endothelial cell apoptosis. In vivo studies showed 1A12 and 2H8 delayed primary tumor growth and reduced spontaneous lung metastasis in breast cancer xenograft model; furthermore, they reduced tumor microvessel density and increased tumor endothelial cell apoptosis. The present study has demonstrated OPN is an effective target in the treatment of tumor, and anti-OPN scFv-Fc fusion proteins could inhibit tumor metastasis by inhibiting angiogenesis and breast cancer cell migration.
引文
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