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目的探讨辛伐他汀对急性髓系白血病(AML)NB4细胞株DNA甲基转移酶(DNMT)的影响及其作用机制。方法将NB4细胞与不同浓度辛伐他汀(终浓度为0、5、10、15μmol/L)共处理。DNA甲基转移酶活性/抑制试验测定DNMT活性,采用实时定量PCR法检测DNMT1、DNMT3A、P53 m RNA的表达水平,采用Western blotting法检测DNM T1、DNM T3A、DNM T3B蛋白水平,流式细胞术检测细胞凋亡。结果辛伐他汀处理后,剂量依赖性的DNMT活性下调(P<0.05);DNMT1、DNMT3A m RNA及蛋白水平降低,P53 m RNA表达水平增高(P<0.05),DNM T3B蛋白表达未见明显变化(P>0.05);辛伐他汀促进NB4细胞凋亡。结论辛伐他汀在NB4细胞中可降低DNMT的活性及表达水平并促进细胞凋亡,有望作为DNMT抑制剂治疗急性白血病。
Objective To investigate the effect and mechanism of simvastatin on DNA methyltransferase (DNMT) in acute myeloid leukemia (AML) NB4 cell line. Methods NB4 cells were co-treated with different concentrations of simvastatin (final concentrations of 0, 5, 10 and 15 μmol / L). DNA methyltransferase activity / inhibition assay was used to detect the activity of DNMT. The expression of DNMT1, DNMT3A and P53 mRNA was detected by real-time quantitative PCR. The protein levels of DNM T1, DNM T3A and DNM T3B were detected by Western blotting, Apoptosis was detected. Results After treatment with simvastatin, DNMT activity was down-regulated (P <0.05); DNMT1, DNMT3A mRNA and protein levels were decreased, P53 mRNA expression was increased (P <0.05), and DNM T3B protein expression was not significantly changed (P> 0.05). Simvastatin promoted the apoptosis of NB4 cells. Conclusion Simvastatin can reduce the activity and expression of DNMT and promote apoptosis in NB4 cells, which is expected to be used as a DNMT inhibitor in the treatment of acute leukemia.