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目的观察霉菌毒素的致癌机制是否与其影响了细胞内的基因修复系统相关。方法体外培养成纤维细胞NIH/3T3细胞系,在培养液中加入16μM的互隔交链孢酚作用后,提取细胞全RNA,进行半定量分析了解其表达量的变化,RT-PCR扩增DNA聚合酶β基因cDNA序列,克隆至T载体后测序,分析其序列变化。结果克隆成功并测序后发现有DNA聚合酶β基因序列的两个位点发生突变。同时,通过半定量分析发现DNA聚合酶β表达量逐渐上调。结论 AOH可导致NIH/3T3细胞中的DNA聚合酶β发生点突变﹑表达量上调。
Objective To investigate whether the carcinogenic mechanism of mycotoxins is related to its influence on intracellular gene repair system. Methods NIH / 3T3 fibroblasts were cultured in vitro. After adding 16μM interleukinol into the culture medium, total RNA was extracted and analyzed by semi-quantitative analysis. The expression of DNA was amplified by RT-PCR The cDNA sequence of polymerase β gene was cloned into T vector and sequenced to analyze its sequence changes. Results After cloning was successful and sequenced, two sites of DNA polymerase β gene sequence were found to be mutated. Meanwhile, the expression of DNA polymerase β was gradually increased by semi-quantitative analysis. Conclusion AOH can cause point mutation of DNA polymerase β in NIH / 3T3 cells and its expression level is up-regulated.