血管内皮生长因子-C反义寡核苷酸对胰腺癌淋巴管及血管生成的影响

Effect on lymphangiogenesis and angiogenesis of pancreatic cancer with antisense oligonucleotide of vascular endothelial growth factor C

目的观察血管内皮生长因子(VEGF)-C反义寡核苷酸对胰腺癌裸鼠原位种植瘤模型淋巴管及血管生成的影响。方法建立人胰腺癌细胞株panc-1裸鼠原位种植瘤模型,将动物随机分为磷酸盐缓冲液(PBS)对照组(A组)、错义对照组(B组)和反义VEGF-C干预组(C组),每组10只,寡核苷酸用量为每次10 mg/kg体重,隔日1次,每周3次,共3周。建模4周后,处死动物,留取血清和瘤体标本,采用酶联免疫吸附试验(ELISA)、免疫组织化学染色法检测反义VEGF-C干预对VEGF-C分泌水平及种植瘤淋巴管和血管生成的影响。结果A、B和C组血清VEGF-C的蛋白表达水平分别为(237．5±41．5)、(221．5±52．3)、(108．6±14．9)ng/L,C组较A、B组明显为低(P＜0．01)；3组种植瘤内淋巴管密度分别为13．8±2．1、12．4±1．9和4．2±1．6,C组较A、B组显著减少(P＜0．01)；3组种植瘤内微血管密度分别为27．5±8．7、25．9±4．2和19．4±5．6,组间差异无统计学意义(P＞0．05)。结论反义寡核苷酸干预可以显著降低胰腺癌裸鼠原位种植瘤模型VEGF-C的表达水平,并对其淋巴管生成具有抑制作用。

Objective To investigate the effect on lymphangiogenesis and angiogenesis of orthotopic implantation of pancreatic cancer with antisense oligonucleotide （ASODN） of vascular endothelial growth factor C（VEGF-C）. Methods Antisense and scamble oligonucleotide of vascular endothelial growth factor C were constructed. Establishment of the model in nude mice with orthotopic implantation for the human pancreatic cancer cell line panc-1. Thirty nude mices were randomized into 3 groups： PBS control group（group A）,scramble control group（group B） and antisense group（group C）. All nude mices were treated once 2 days as 3 times per week, for 3 weeks（oligonucleotide 10mg/kg/time）. After treatment was completed, the exocrine of VEGF-C and lymphangiogenesis and angiogenesis of pancreatic cancer were detected by ELISA and immunohistochemistry. Results With treatment of ASODN, the exocrine of VEGF-C was inhibited significantly in the serum （P 〈 0.05）. The levels were （237.5 ± 41.5 ）, （221.5± 52.3） and （108.6 ± 14.9） ng/L in group A, B and C respectively. Meanwhile, the lymph vessel density was decreased significantly （P 〈 0.05）, 13.8 ± 2.1,12.4 ± 1.9 and 4.2 ± 1.6 in each group respectively. Though the microvessel densities were tended to decrease, 27.5 ±8.7, 25.9 ±4.2 and 19.4 ± 5.6 in each group respectively, but there wre not different statisticaly （P 〉 0.05）. Conclusion The antisense oligonucleotide of VEGF-C decrese the level of VEGF-C in nude mice of orthotopic implantation of pancreatic cancer, and have inhibition effect on lymphangiogenesis.