VEGF—C反义多肽对人肝癌细胞HepG2增殖及侵袭的抑制效应

Anti-tumor effect of antisense peptide of VEGF-C on human hepatic cancer cells HepG2

目的检测血管内皮生长因子-C（VEGF—C）反义多肽对人肝癌细胞增殖和侵袭能力的影响。方法制备VEGF-C的反义多肽，通过与肝癌细胞株HepG2共培养，用四甲基偶氮唑盐微量酶反应比色法（MTY）检测细胞增殖率，人工基底膜侵袭实验检测细胞侵袭能力；构建肝癌细胞HepG2裸鼠皮下移植瘤模型，瘤体内注射YEGF—C反义多肽，观察其对肝癌细胞的抑制作用；WesternBlot检测VEGF-C在瘤组织的表达情况；免疫组化分析肿瘤组织VEGFR-3阳性脉管密度。结果VEGF—C反义多肽具有较强的生物学活性，对肝癌细胞具有明显增殖抑制作用（P〈0．05）；25、50、100μg／mlVEGF-C反义多肽对侵袭重组基底膜的抑制率分别为41．7％、57．3％和66．9％（P〈0．05）；瘤组织内给予VEGF-C反义多肽后，肿瘤生长受抑制（P〈0．05），WesternBlot分析VEGF—C在瘤组织中的表达降低（P〈0．05）；免疫组化分析VEGF-C反义多肽可抑制肿瘤组织YEGFR-3阳性脉管形成（P〈0．05）。结论制备出的VEGF-C反义多肽具有良好的生物学活性，通过抑制肝癌细胞的增殖、侵袭能力及肿瘤组织VEGFR-3阳性脉管形成而发挥抗肝癌作用。

Objective To investigate the anti-tumor effects of the antisense peptide of vascular endothelial growth factor（VEGF）-C on proliferation and invasion ability of human hepatic cancer cell. Methods Antisense peptide of VEGF-C was prepared, hepatic cancer cell line HepG2 was treated by antisense peptide of VEGF-C, the cell-growth rate was evaluated by MTT method and the invasive capacity was evaluated by reconstituted basement membrane invasion assay. The model of subcutaneous tumor was generated by injection of HepG2 cells into the nude mice, antisense peptide of VEGF- C was subcutaneously injected around xenograft tumors after tumor cell innoculation. Tumor growth was observed, VEGF-C expression was measured by western blot and formation of VEGFR-3 positive vessels in tttmor tissues was analyzed by immunohistochemical analysis. Results VEGFC antisense peptide effectively inhibited HepG2 cells growth（ P 〈0. 05）, suppression ratio of HepG2 cells invasion was 41.7% ,57. 3% and 66. 9% according to VEGF-C antisense peptide concentration of 25,50 and 100 μg/ml （ P 〈0. 05）. Tumor growth was significantly inhibited in mice injected with antisense peptide of VEGF-C （ P 〈0. 05）. Western blot analysis showed expression of VEGF-C protein was also inhibited（ P 〈0. 05） ; nalysis demonstrated significant reduction of VEGFR-3 positive vessel formation in tumor tissues（ P 〈0. 05）. Conclusion Antisense peptide of VEGF-C shows effective biological activity, and it can inhibit tumor growth through inhibiting tumor cell proliferation, suppressing tumor invasion and inhibiting VEGFR-3 positive vessel formation.