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目的探讨新疆雪莲总黄酮对DNA甲基化酶活性及癌细胞DNA甲基化状态的影响。方法采用DNA甲基化酶活性分析实验检测雪莲总黄酮对DNA甲基化酶活性的影响,甲基化特异性PCR(MSP)检测雪莲总黄酮处理2d后食管癌KYSE510细胞抑癌基因P16启动子CpG岛甲基化状态;同时流式细胞仪检测细胞凋亡。结果雪莲总黄酮抑制DNA甲基化酶M.Alu I活性,MSP实验结果显示雪莲总黄酮处理2 d后,癌细胞未甲基化状态P16基因的表达水平升高,KYSE510细胞P16基因mRNA恢复表达。流式分析结果显示,雪莲总黄酮处理KYSE510癌细胞2 d后,处理组细胞凋亡率与对照组比较显著增加(P﹤0.05)。结论雪莲总黄酮可以通过影响DNA甲基化酶活性,激活表观沉默的抑癌基因表达,抑制癌细胞增殖。
Objective To investigate the effects of total flavonoids from Xinjiang snow lotus on DNA methylation and DNA methylation status of cancer cells. Methods DNA methyltransferase activity assay was used to detect DNA methylase activity. Methylation-specific PCR (MSP) was used to detect the expression of the suppressor gene P16 promoter of KYSE510 cells CpG island methylation status; meanwhile, flow cytometry was used to detect apoptosis. Results Total flavonoids of Elaeagnus nodosii inhibited the activity of DNA methylase M. Alu I. The results of MSP showed that the expression of P16 gene in unmethylated cancer cells increased and the expression of P16 gene in KYSE510 cells recovered after 2 days . Flow cytometry analysis showed that the apoptosis rate of KYSE510 cancer cells treated with total flavonoids of Saussurea increased significantly (P <0.05) compared with that of the control group. Conclusion The total flavones of Saussurea can inhibit the proliferation of cancer cells by affecting the DNA methylase activity, activating the expression of silencing tumor suppressor genes.