Ephrin-A1/Fc融合蛋白对人肝癌细胞系Huh-7生物学特性的影响

目的 探讨Ephritr-A1基因在肝细胞癌发生、发展以及血管生成中的作用.方法 人肝癌细胞系Huh-7分别经Ephrin-A1/Fc融合蛋白和IgG/Fc作用后,MTT法绘制细胞生长曲线,细胞基质黏附实验检测对细胞黏附能力的影响,Transwe11法检测细胞侵袭能力的改变.通过裸鼠移植瘤模型的建立检测Ephrin-A1/Fc融合蛋白对Huh-7细胞体内致瘤能力的影响,并计算瘤体的微血管密度(MVD),判断其对血管生成的影响.结果 Huh-7细胞的黏附率为(131.25±9.57)%,细胞侵袭实验显示Ephrin-A1/Fc融合蛋白作用后,穿过滤膜的细胞数.明显多于IgG/Fc组和空白对照组,其差异均有统计学意义(P<0.05);裸鼠成瘤实验结果 显示,6周后Ephrin-A1/Fc组裸鼠体积为(186.35±16.24)mm3,其差异均有统计学意义(P<0.05);而瘤体内的MVD值Ephrin-A1/Fc也明显高于IgG/Fc组和空白对照组,其差异均有统计学意义(P<0.05).Ephrin-A1/Fc融合蛋白对Huh-7细胞并无增殖促进作用(P>0.05).结论 Ephrin-A1基因在肝细胞癌的侵袭及血管生成的过程中发挥重要的作用,因此有望成为肝细胞癌基因治疗新的靶点.

Effect of Ephrin-A1/Fc fusion protein on biological features of human hepatocellular carcinoma cell line Huh-7

Objective To study the role of Ephrin-A1 gene in tumorigenesis, development and angiogenesis of hepatocellular carcinoma (HCC). Methods MTT, cell-matrix adhesive assay and tr-answell cell assay were used to test the effect of Ephrin-Al/Fc fusion protein on the proliferation, ad-hesive and invasive ability of Huh-7 cell, respectively. In in-vivo study, the growth of Huh-7 xeno-transplant was observed in BALB/C nude mice. The effect of Ephrin-Al/Fc fusion protein on Huh-7's tumorigenesis ability was evaluated, and the tumor microvessel density(MVD) was determined to esti-mate the effect of Ephrin-Al/Fc fusion protein on angiogenesis of HCC. Results Ephrin-Al/Fc fu-sion protein improved the adhesive ability of Huh-7 cell. At 120 min, the adhesive rate of Ephrin-Al/ Fc group was (131.25±9.57)% and that of the IgG/Fc group was (95.18±15.55)%. There was significant difference between the 2 groups (P<0.05). Tumor invasion assay showed that the number of invaded Huh-7 cells in Ephrin-Al/Fc group(77.25±7.95) was markedly higher than that in IgG/ Fc group (62.00±6.97) and control group (47.45±3.09) 24 h later(P<0.05). In in-vivo study, it was found that the xenograft volume of Ephrin-Al/Fc group(186.35±16.24) mm3 was higher than that of the IgG/Fc group (92.31±26.89)mm3 and control group (102.73±46.77) mm3 (P<0.05). The MVD value of Ephrin-Al/Fc group (18.25±7.35) was remarkably higher than that of the IgG/ Fc group (12.56±6.86) and control group(10.33±5.09) (P<0.05). Ephrin-Al/Fc fusion protein did not improve the proliferation of Huh-7 (P>0.05). Conclusion Ephrin-A1 gene plays an impor-tant role in tumorigenesis, development and angiogenesis of HCC. Therefore, it can be used as the new target of the gene therapy of HCC.