[地西他滨对白血病细胞中DKK1基因去甲基化的影响]
[Effect of Decitabine on DKK1 Gene Demethylation in Leukemia Cells].
作者信息
Liu Feng-Zhi, He Ling, Wang Ji-Shi, Zhang Song, Zhu Hong-Qian
机构信息
Department of Hematology, Guizhou Provincial People's Hospital, Guiyang 550002, Guizhou Province, China.
Department of Hematology, Affiliated Hospital of Guiyang Medical University, Guiyang 550002, Guizhou Province, China.
出版信息
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2016 Feb;24(1):56-60. doi: 10.7534/j.issn.1009-2137.2016.01.011.
OBJECTIVE
To explore the effect of decitabine on Dickkopf-1 (DKK1) gene expression level and its downstream Wnt signaling pathway in acute myeloid leukemia (AML) cell line HL-60.
METHODS
Flow cytometry and DNA ladder analysis were performed to detect apoptosis in HL-60 cell treated with different concentration of decitabine. Methylation-specific polymerase chain reaction (MS-PCR) was used to examine the methylation status of DKK1 gene. The expressions of mRNA and protein were determined by qRT-PCR and Western blot, respectively.
RESULTS
Flow cytometric detection showed that after treating HL-60 cell line with decitabine of different concentrations for 48 h, the early apoptosis of HL-60 cells increased significantly as compared with control group (P < 0.05). DNA ladder analysis showed that the DNA ladder and demethylation of DKK1 gene appeared. RT-PCR and Western blot showed that the expressions of mRNA and protein increased. The protein expressions of β-catenin and C-MYC decreased.
CONCLUSION
The decitabine can promote the apoptosis of HL-60 cells throngh demethylation of DDK1 gene and inhibition of Wnt signalling pathway.
目的
探讨地西他滨对急性髓系白血病（AML）细胞系HL-60中Dickkopf-1（DKK1）基因表达水平及其下游Wnt信号通路的影响。
方法
采用流式细胞术和DNA梯状分析检测不同浓度地西他滨处理的HL-60细胞的凋亡情况。甲基化特异性聚合酶链反应（MS-PCR）用于检测DKK1基因的甲基化状态。分别通过qRT-PCR和蛋白质免疫印迹法检测mRNA和蛋白质的表达。
结果
流式细胞术检测显示，用不同浓度地西他滨处理HL-60细胞系48小时后，与对照组相比，HL-60细胞的早期凋亡显著增加（P<0.05）。DNA梯状分析显示出现了DKK1基因的DNA梯状条带和去甲基化。逆转录-聚合酶链反应（RT-PCR）和蛋白质免疫印迹法显示mRNA和蛋白质表达增加。β-连环蛋白和C-MYC的蛋白质表达降低。
结论
地西他滨可通过DKK1基因去甲基化和抑制Wnt信号通路促进HL-60细胞凋亡。
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