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目的探讨叶酸和维生素A对MNNG诱导的哈族食管上皮DNMT1高表达细胞PI3K/AKT信号通路的影响。方法在1.500×10~(-5)mol/L MNNG转化浓度、不同浓度叶酸(0.800μg/ml和8.000μg/ml)和维生素A(2.500μg/ml)处理不同时间后,采用PCR和Western blot法检测各干预组细胞PI3K/AKT信号通路关键因子表达水平。结果 (1)MNNG半数抑制浓度为23.897×10~(-5)mol/L,转化剂量为1.500×10~(-5)mol/L;(2)经MNNG、叶酸和维生素A作用细胞一定时间后,(1)随着叶酸浓度增加和维生素A作用可使细胞AKT蛋白表达水平逐渐降低,pp2a和PTEN蛋白表达水平逐渐增加,而随着时间延长,细胞PTEN mRNA和AKT蛋白表达水平逐渐增加,pp2a和PTEN蛋白表达水平逐渐降低;(2)对蛋白表达来说,叶酸和维生素A之间存在交互作用。结论叶酸和维生素A对MNNG致哈族食管上皮DNMT1高表达细胞PI3K/AKT信号通路AKT、pp2a和PTEN基因表达具有一定的干预作用。
Objective To investigate the effect of folic acid and vitamin A on the PI3K / AKT signaling pathway in DNMT1 overexpressing cells of Ha ethnic esophageal epithelium induced by MNNG. Methods After treated with MNNG at a concentration of 1.500 × 10 ~ (-5) mol / L and different concentrations of folic acid (0.800μg / ml and 8.000μg / ml) and vitamin A (2.500μg / ml) Method to detect the expression of PI3K / AKT signaling pathway key factor in each intervention group. Results (1) The half maximal inhibitory concentration of MNNG was 23.897 × 10 -5 mol / L, and the conversion dose was 1.500 × 10 -5 mol / L. (2) The effect of MNNG, folic acid and vitamin A on cells for a certain period of time (1) With the increase of folic acid concentration and the action of vitamin A, the expression of AKT protein in cells gradually decreased and the expression levels of pp2a and PTEN protein gradually increased. With the prolongation of time, the expression of PTEN mRNA and AKT protein gradually increased, pp2a and PTEN protein levels gradually decreased; (2) for the protein expression, there is interaction between folic acid and vitamin A. Conclusions Folic acid and vitamin A can interfere with the expression of PI3K / AKT signaling pathway AKT, pp2a and PTEN in MNNG-induced DNMT1 overexpressing cells.