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目的 研究血管内皮生长因子 (VEGF)反义寡核苷酸 (ODN)是否可以抑制人肝癌细胞系VEGF的蛋白表达 ,进而抑制肝癌的血管形成 ,探讨肝癌基因治疗的新方法。方法 人肝癌细胞系 772 1能表达VEGF ,其培养上清液对牛主动脉内皮细胞 (BAEC)的生长有促进作用。采用人工合成反义、正义ODN分别加入 772 1细胞培养液 ,比较 772 1细胞VEGF的表达以及各上清液刺激内皮细胞生长的受抑情况 ,即可了解ODN通过抑制VEGF的表达抑制肝癌的血管形成情况。结果 VEGF反义ODN明显抑制了人肝癌细胞VEGF的表达 (灰度值 :反义组 12 2 6 ,正义组 10 1 3 ,对照组 10 6 3,P <0 0 1) ,且抑制率与剂量呈线性关系 (当剂量为 1、2 5、5、10 μmol/L时抑制率分别为 :6 3 2 %、82 4%、2 10 0 %、2 87 5 % )。结论 VEGF反义ODN能够抑制肝癌的血管形成 ,其有望成为抗肝癌的新一代基因治疗手段
Objective To investigate whether vascular endothelial growth factor (VEGF) antisense oligonucleotide (ODN) can inhibit the protein expression of VEGF in human hepatocellular carcinoma cell lines, thereby inhibiting hepatic angiogenesis, and explore new methods for gene therapy of hepatocellular carcinoma. Methods The human hepatocellular carcinoma cell line 772 1 was able to express VEGF and its culture supernatant promoted the growth of bovine aortic endothelial cells (BAEC). Using artificially synthesized antisense and sense ODN to add 772 1 cell culture fluid, comparing the expression of VEGF in 772 1 cells and the suppression of endothelial cell growth inhibition by each supernatant, we can know that ODN inhibits the blood vessels of liver cancer by inhibiting the expression of VEGF. Formation situation. RESULTS: VEGF antisense ODN significantly inhibited the expression of VEGF in human hepatoma cells (gray value: antisense group 1226, sense group 1013, control group 106O3, P<0 01), and the inhibition rate and dose The relationship was linear (inhibition rates were 632,82,210, 2875%) at doses of 1, 25, 5, and 10 μmol/L, respectively. Conclusion VEGF antisense ODN can inhibit the angiogenesis of liver cancer, which is expected to become a new generation of gene therapy for liver cancer.