反义寡核苷酸抑制肝癌细胞血管内皮生长因子的表达

Inhibitory effect of antisense oligodeoxynucleotide on the expression of vascular endothelial growth factor in human hepatocellular carcinoma

目的 :研究血管内皮生长因子 (VEGF)反义寡核苷酸 (ODN)对人肝癌细胞系VEGF表达的抑制作用 ,探讨肝癌基因治疗的新方法。方法 :人肝癌细胞系SMMC772 1能表达VEGF ,采用人工合成反义、正义ODN分别作用于SMMC772 1细胞 ,RT -RCR、流式细胞仪比较各处理组VEGFmRNA及其蛋白的表达变化情况。结果 :VEGFRT -PCR扩增条带的图像分析平均灰度值显示 ,反义组为 0 4 379,正义组为 0 736 7,对照组为 0 74 6 2。流式细胞仪分析VEGF蛋白表达率 ,反义组为 17 2 % ,正义组为 31 8% ,对照组为 31 2 %。结论 :反义ODN能够抑制人肝癌细胞的表达 ,其有望成为抗肝癌的新一代基因治疗药物。

Objective:To investigate the inhibitory effect of antisense oligodeoxynucleotide(ODN)on the expression of vascular endothelial growth factor(VEGF)to explore a new gene therapy for human hepatocellular carcinoma.Methods:Synthetic antisense and sense ODNs were added into the medium of SMMC7721 cells that can express VEGF.Reverse-transcriptase polymerase chain reaction(RT-PCR)and flow cytometry(FCM)were employed make a comparison of the changes in the expressions of mRNA and potein in VEGF among antisense,sense and control group.Results:RT-PCR showed that the average gray levels in antisense,sense and control group were 0.4379,0.7367,and 0.7462,respectively.FCM showed that the expression rates of protein in VEGF in antisense,sense and control group were 17.2%,31.8%,and 31.2%,respectively.Conclusion:Antisense ODN can inhibit the expression of VEGF in human hepatocellular carcinoma,which is hoped to be a new gene therapeutic drug for hepatocellular carcinoma.