人源性抗bFGF抗体抑制人黑色素瘤生长的作用研究

The effect of human antibody against basic fibroblast growth factor on the growth of human melanoma

目的研究人源性抗bFGF抗体对人黑色素瘤A375细胞的体内外增殖抑制作用。方法体外培养人黑色素瘤A375细胞,CCK-8法检测人源性抗bFGF抗体对A375细胞增殖的影响;Western blot检测人源性抗bFGF抗体对bFGF下游信号通路中ERK1/2磷酸化的影响;建立裸鼠皮下移植瘤模型,通过记录各处理组裸鼠皮下移植瘤的体积、质量及裸鼠体质量变化,研究人源性抗bFGF抗体在体内对人黑色素瘤生长的抑制作用;免疫组化检测肿瘤组织内的微血管密度。结果 CCK-8实验结果表明人源性抗bFGF抗体能抑制A375细胞的生长,在800 ng/ml的质量浓度下,抑制率为24%;Western blot结果表明人源性抗bFGF抗体显著抑制FGFR下游信号通路中ERK1/2的磷酸化;裸鼠体内试验中,人源性抗体明显抑制了肿瘤的生长,抑制率为28.12%。免疫组化结果显示各组肿瘤组织中微血管密度均有下降。结论该株人源性抗体在体外可有效的抑制人黑色素瘤细胞的生长,并能在体内抑制肿瘤生长及血管新生。

This study designed to investigate the inhibitory effects of human anti-bFGF antibody on human melanoma in vitro and in vivo. Firstly the proliferation inhibition of human melanoma cells by human anti-bFGF antibody was analyzed by CCK-8, and the phosphorylation level of ERK was detected by Western blot. Xenograft nude mouse model was established to study the inhibitory effects of human anti-bFGF antibody on melanoma in vivo; microvessel density （MVD） of the tumor tissue was analyzed by immunohistochemistry. The results of the proliferation inhibition showed that the proliferation of melanoma cells could be inhibited by human anti-bFGF antibody, and the inhibition rate was about 24% at the antibody concentration of 800 ng/ml; Western blotting results showed that ERK1/2 phosphorylation was inhibited by human anti-bFGF antibody, which demonstrated that human anti-bFGF antibody could inhibit the proliferation of tumor cells by inhibiting the ERK signal pathway. The tumor xenografts nude mice model showed that human anti-bFGF antibody could inhibit the tumor growth in vivo, and the inhibition rate was about 28.12%. Furthermore, the microvessle density of tumor tissue was decreased obviously. These results demonstrated that human anti-bFGF antibody could inhibit the proliferation of human melanoma cell in vitro and inhibit tumor growth and angiogenesis in vivo.