CAJ | 학술논문

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、20 μmol/L的VEGF ASODN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VEGF mRNA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:VEGF ASODN 3个浓度组(5、10和20 μmol/L)处理的Namalwa细胞VEGF mRNA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20 μmol/L VEGF ASODN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:VEGF ASODN在体外能够抑制Namalwa细胞VEGF的表达.
위료연구류대린산화수식적혈관내피생장인자(vascular endothelial growth factor,VEGF)반의과핵감산(antisense oligodeoxynucleotide,ASODN)대인림파류세포계Namalwa세포VEGF표체적영향,장종농도분별위5、10、20 μmol/L적VEGF ASODN화착의서렬여인림파류세포계Namalwa세포분별부육24、48소시,채용RT-PCR검측VEGF mRNA적표체,채용련매균항생소단백-과양화매면역조직화학법(streptavidin/peroxidase,SP법)검측VEGF적표체.결과표명:VEGF ASODN 3개농도조(5、10화20 μmol/L)처리적Namalwa세포VEGF mRNA적표체분별위1.38、0.96、0.57,착의서렬조화대조조분별위1.79、1.84.당가입20 μmol/L VEGF ASODN작용48소시후,세포내VEGF단백수평현저감소,이착의서렬조Namalwa세포VEGF단백수평미견명현개변.결론:VEGF ASODN재체외능구억제Namalwa세포VEGF적표체.
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 (5,10,20 μmol/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.