Ephrin—Al／Fc融合蛋白对人肝癌细胞系Huh-7生物学特性的影响

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

目的探讨Ephrin-Al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—Al／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，Transwell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-Al／Fc融合蛋白对Huh一7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Huh7细胞的黏附率为（131．25±9．57）％，细胞侵袭实验显示Ephrin-Al／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〈O．05）；裸鼠成瘤实验结果显示，6周后Ephrin-Al／Fc组裸鼠体积为（186．35±16．24）mm^3，其差异均有统计学意义（P〈O．05）；而瘤体内的MVD值EphrinAl／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-Al／Fc融合蛋白对Huh-7细胞并无增殖促进作用（P〉O．05）。结论Ephrin—Al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。

Objective To study the role of Ephrin-Al gene in tumorigenesis, development and angiogenesis of hepatocellular carcinoma （HCC）. Methods MTT, cell-matrix adhesive assay and transwell 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 estimate the effect of Ephrin Al/Fc fusion protein on angiogenesis of HCC. Results Ephrin Al/Fc fusion 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/Fe 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-A1/Fc group（186.35 ±16.24） mm^3 was higher than that of the IgG/Fc group （92.31±26.89）mm3 and control group （102.73±46.77） mm^3 （P〈0.05）. The MVD value of Ephrin-A1/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-Al gene plays an important role in tumorigenesis, development and angiogenesis of HCC. Therefore, it can be used as the new target of the gene therapy of HCC.