摘要
目的探讨叶酸在N-甲基-N-硝基-N-亚硝基胍(MNNG)致哈萨克族食管上皮细胞APE1、MGMT基因mRNA及蛋白表达中的作用。方法采用3×3析因设计将体外培养的哈萨克族食管上皮细胞暴露于不同浓度叶酸(0.00、0.40和0.80μg/ml)和MNNG(0.00、0.75和1.50μg/ml)的培养液中作用48、72和96 h后,采用RT-PCR法检测APE1、MGMT基因mRNA的表达水平,采用Western Blot法检测APE1、MGMT基因蛋白表达水平。结果随着叶酸浓度降低和MNNG浓度增加,APE1基因的mRNA及蛋白表达水平随着损伤的增加而增加,差异有统计学意义(P<0.01);MGMT基因的mRNA及蛋白表达水平则随损伤的增加而下降,差异有统计学意义(P<0.01)。结论叶酸浓度降低对MNNG致哈萨克族食管上皮细胞损伤有一定的促进作用,而保持充足的叶酸则对MNNG的损害起到一定的保护作用。
Objective To investigate the role of folie acid in MNNG induced APE1, MGMT mRNA and protein expression for Kazakh normal esophageal epithelial cell. Methods Kazakh esophageal epithelial cells cultured in vitro were exposed to medium containing different concentrations of folic acid (0. 00, 0. 40, 0. 80 μg/ml) and MNNG (0. 00, 0.75, 1.50 μg/ml) by using 3 x 3 factorial design trial. Some different indexes were detected in 48 h, 72 h and 96 h. The mRNA and protein levels of APE1 and MGMT were tested by RT- PCR and Western-Blot. Results With the decrease of the concentration of folic acid and the increases of the concentration of MNNG, the following result showed that the mRNA expression and protein expression of APE1 in Kazakh esophageal epithelial cells were significantly increased (P〈0.01).The expression of mRNA and protein of MGMT gene were significantly decreased (P 〈0.01). Conclusion The reduction of the concentrations of folic acid promotes the Kazakn esophageal epithelial cells' damage caused by MNNG. Sufficient folic acid plays a protective role in the damage caused by MNNG.
出处
《毒理学杂志》
CSCD
北大核心
2017年第6期437-441,共5页
Journal of Toxicology
基金
国家自然科学基金(81460502)
新疆维吾尔自治区科技厅自然科学基金(2015211C032)