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目的探讨黄曲霉毒素G1(AFG1)对体外培养的人食管鳞状上皮细胞表面HLA-Ⅰ分子表达的影响。方法采用原代培养、免疫印迹(Western blot)和反转录聚合酶链反应(RT-PCR)分析方法研究不同浓度AFG1(50、100、1000和2000μg/L)处理后人食管鳞状上皮细胞表面HLA-Ⅰ分子表达的变化。结果免疫印迹结果表明,经不同浓度AFG1(50、100、1000、2000μg/L)处理细胞24h后,100、1000、2000μg/L AFG1处理组HLA-ABC的蛋白表达均明显低于溶剂对照组(P<0.05),而且在100～2000μg/L浓度范围内,人食管鳞状上皮细胞表面HLA-Ⅰ分子的表达逐渐降低,两者呈明显的负相关关系(r=-0.921,n=3,P<0.01)。RT-PCR检测在mRNA水平进一步证实了AFG1对HLA-ABC表达的影响。其中100、1000、2000μg/L AFG1处理细胞后,HLA-A mRNA的表达明显低于溶剂对照组(P<0.05);2000μg/L处理组HLA-B mRNA的表达较溶剂对照组降低(P<0.05);而各AFG1处理组HLA-C mRNA的表达没有明显影响。结论 AFG1处理可以抑制人食管鳞状上皮细胞表面HLA-Ⅰ分子的表达。
Objective To investigate the effect of aflatoxin G1 (AFG1) on the expression of HLA-Ⅰ on the surface of human esophageal squamous epithelial cells cultured in vitro. Methods Primary culture, Western blot and reverse transcription polymerase chain reaction (RT-PCR) were used to study the effects of different concentrations of AFG1 (50, 100, 1000 and 2000 μg / L) on human esophageal squamous epithelial cells Changes of surface HLA-Ⅰ molecule expression. Results The results of Western blotting showed that after treated with different concentrations of AFG1 (50, 100, 1000 and 2000 μg / L) for 24 h, the protein expression of HLA-ABC in AFG1 treated group was significantly lower than that in the solvent control group The expression of HLA-Ⅰ on the surface of human esophageal squamous epithelial cells gradually decreased in the range of 100-2000 μg / L with a significant negative correlation (r = -0.921, n = 3, P <0.05) P <0.01). The effect of AFG1 on HLA-ABC expression was further confirmed by RT-PCR at the mRNA level. The expression of HLA-A mRNA in AFG1 cells treated with 100, 1000, 2000μg / L AFG1 was significantly lower than that in the solvent control group (P <0.05). The expression of HLA-B mRNA in 2000μg / L treatment group was lower than that of the solvent control group 0.05). However, the expression of HLA-C mRNA in AFG1-treated group had no significant effect. Conclusion AFG1 treatment can inhibit the expression of HLA-Ⅰ on the surface of human esophageal squamous cell.