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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 effect of antisense oligodeoxynucleotide (ASODN) modified by phosphorothioated modified vascular endothelial growth factor (VEGF) on the expression of VEGF in human lymphoma cell line Namalwa cells, the final concentrations were The VEGFASODN of 5, 10 and 20μmol / L and the missense sequence were respectively incubated with the human lymphoma cell line Namalwa for 24 and 48 hours. The expression of VEGFmRNA was detected by RT-PCR. The expression of VEGFmRNA was detected by streptavidin-peroxidase immunohistochemistry Method (streptavidin / peroxidase, SP method) to detect the expression of VEGF. The results showed that the expression of VEGFmRNA in Namalwa cells treated with VEGFASODN3 (5, 10 and 20μmol / L) were 1.38,0.96 and 0.57, respectively. The missense sequence and control group were 1.79 and 1.84, respectively. When adding 20μmol / LVEGFASODN for 48 hours, the level of VEGF protein decreased significantly, while the missense sequence of Namalwa cells VEGF protein levels did not change significantly. Conclusion: VEGFASODN can inhibit the expression of VEGF in Namalwa cells in vitro.