反义寡核苷酸对人淋巴瘤细胞系Namalwa细胞血管内皮生长因子表达影响的体外研究(英文)

Effect of Antisense Oligodeoxynucleotides on the Expression of Vascular Endothelial Growth Factor in Namalwa cell In Vitro

为了研究硫代磷酸化修饰的血管内皮生长因子(vascular endothelial growth factor,VEGF)反义寡核苷酸(antisense oligodeoxynucleotide,ASODN)对人淋巴瘤细胞系Namalwa细胞VEGF表达的影响,将终浓度分别为5、10、20μmol/L的VEGFASODN和错义序列与人淋巴瘤细胞系Namalwa细胞分别孵育24、48小时,采用RT-PCR检测VEGFmRNA的表达,采用链酶菌抗生素蛋白-过氧化酶免疫组织化学法(streptavidin/peroxidase,SP法)检测VEGF的表达。结果表明:VEGFASODN3个浓度组(5、10和20μmol/L)处理的Namalwa细胞VEGFmRNA的表达分别为1.38、0.96、0.57,错义序列组和对照组分别为1.79、1.84。当加入20μmol/LVEGFASODN作用48小时后,细胞内VEGF蛋白水平显著减少,而错义序列组Namalwa细胞VEGF蛋白水平未见明显改变。结论:VEGFASODN在体外能够抑制Namalwa细胞VEGF的表达。

In order to study the effects of phosphorothioated antisense oligodeoxynucleotides （ASODN） on the expression of VEGF in human lymphoma cell line Namalwa cells, human lymphoma cell line Namalwa cells were incubated with VEGF ASODN （the final concentrations of VEGF ASODN were 5, 10, 20 μmol/L respectively ）, or scrambled sequence for 24 or 48 hours. The expressions of VEGF mRNA and VEGF protein were detected by reverse transcriptase-polymerase chain reaction and streptavidin peroxidase （SP） immunohistochemistry respectively. The results showed that the expression levels of VEGF mRNA in Namalwa cells treated with three concentration levels （5,10, 20μmol/L of ASODN） were 1.38, 0.96 and 0.57 respectively. Those in PBS-treated cells and scrambled sequence treated cells were 1. 79 and 1. 84. When treated with 20 μmol/L VEGF ASODN for 48 hours, VEGF protein of Namalwa cells decreased greatly. Meamwhile, there was no obvious change in the scrambled sequence treated group. It is concluded that VEGF ASODN can suppress the VEGF expression in Namalwa cells in vitro.