亚硝基乙酰青霉胺对腺样囊性癌细胞VEGF表达双向调控作用

摘要

Objective:To observe the effect of S-nitroso-N-acetyl-penicillamine (SNAP) on the regulation of VEGF mRNA levels in Adenoid cystic carcinoma of slavery gland tumor(ACCs) cells. Method: A concentration gradient of SNAP (31.25μmol/L、62.25μmol/L、125μmol/L、250μmol/L、500μmol/L) was used to stimulate ACC-M and ACC-2 cells to identi-fy the expression curve of VEGF mRNA levels. Then, after pretreated with 100ng/ml LPS for 12 hours, the ACCs cells were stimulated by 500μmol/L SNAP on 0, 1, 2, and 4 hours, respectively. Cell Counting Kit-8 (CCK-8) was used to analyze the toxicity of SNAP on ACCs cells, and semi-quantitative RT-PCR was used to detect the VEGF mRNA levels. Results:SNAP significantly increased the VEGF mRNA level in the concentration of 31.25μmol/L, while effectively suppressed the VEGF mRNA level in the concentration of 500μmol/L on a time dependent manner. Conclusion:It was suggested that SNAP might exhibit bidirectional regulation function on modulating VEGF expression in ACCs cells.%目的：观察亚硝基乙酰青霉胺（SNAP）对腺样囊性癌细胞中VEGF表达水平的调控作用。方法：将31.25μmol/L、62.25μmol/L、125μmol/L、250μmol/L、500μmol/L不同浓度的SNAP作用于ACC-M和ACC-2细胞，或用浓度为100ng/mL的LPS预刺激12h后，用500μmol/L的SNAP分别刺激ACCs细胞0、1、2、4小时。采用CCK-8活细胞计数试剂盒对ACCs细胞进行毒性分析，半定量RT-PCR检测各组细胞中VEGF表达水平。结果：（1）在0μmol/L~500μmol/L浓度的SNAP作用下，细胞活力均保持在88%；用500μmol/L的SNAP分别作用不同时间ACC-M和ACC-2细胞活力，其活力均保持在86%。（2）当SNAP的刺激浓度在31.25μmol/L时能显著上调VEGF的表达，而500μmol/L的SNAP则显著下调ACCs细胞中VEGF的mRNA水平（P<0.01），仅为基础mRNA水平的0.23倍和0.17倍。结论：在涎腺腺样囊性癌细胞中亚硝基乙酰青霉胺对血管内皮生长因子的表达存在着双向调控作用。