脂质体介导的VEGF—C反义寡核苷酸对肺癌微淋巴管及微血管生成的影响

Effects on lymphangiogenesis and angiogenesis of lung cancer with antisense oligonudeotides of vascular endothelial growth factor C

目的观察脂质体介导的血管内皮生长因子C（VEGF-C）反义寡核苷酸对肺癌裸鼠皮下原位种植瘤模型微淋巴管及微血管生成的影响。方法建立BALB／C小鼠皮下肺癌模型30只，随机分为磷酸盐缓冲液对照组、正义寡核苷酸对照组（SODN组）和反义寡核苷酸干预组（AODN组）。接种人肺腺癌细胞株A549后24h内，用PBS、SODN及AODN分别行皮下注射，观察小鼠肿瘤的生长情况。建模4周后，处死动物，留取瘤体标本，采用RT-PCR、Western Blot和免疫组织化学方法检测VEGF-C反义寡核苷酸对种植瘤中VEGF-C表达水平和微淋巴管及微血管生成的影响。结果种植瘤内VEGF-C mRNA和蛋白表达量AODN组低于对照组（P〈0．05），种植瘤内微淋巴管密度AODN组亦较对照组减少（P〈0．01）。种植瘤内微血管密度各组间差异无统计学意义（P〉0．05）。结论脂质体介导的VEGF-C反义寡核苷酸可以显著降低肺癌裸鼠皮下原位种植瘤模型VEGF—C的表达水平，并对其微淋巴管生成具有较强的抑制作用。

Objective To investigate the effects on lymphangiogenesis and angiogenesis of orthotopic implantation of lung cancer in nude mice with antisense oligonucleotides of VEGF- C. Methods The model in nude mice was established with orthotopic implantation for the human lung cancer cell line A549. Thirty nude mice were randomized into three groups： PBS control group, sense oligonucleotides control group and antisense oligonucleotides group （ AODN group）. After treatments were completed, the expression of VEGF-C and lymphatic microvessel density （LMVD） and microvessel density （MVD） of lung cancer were detected by RT-PCR, Western Blot and immunohistochemistry. Results The expression of VEGF-C in AODN group was inhibited significantly （ P 〈 0. 05 ）. The LMVD in AODN group was decreased significantly （ P 〈 0. 01 ）. Though the MVD in AODN group was also decreased, but there were no significant differences compared with control groups （ P 〉 0.05 ）. Conclusions The antisense oligonucleotides of VEGF-C can inhibit the expression of VEGF-C in nude mice of orthotopic implantation of lung cancer. It could inhibite the lymphangiogenesis.