青黄散联合健脾补肾方对骨髓增生异常综合征患者DACT1基因甲基化及Wnt/β-catenin通路的影响

Effects of Qinghuang Powder combined with Jianpi Bushen Formula on DACT1 gene methylation and Wnt/β-catenin pathway in myelodysplastic syndrome patients

目的:探讨青黄散联合健脾补肾方对骨髓增生异常综合征(MDS)患者DACT1基因甲基化及Wnt/β-catenin通路的影响。方法:亚硫酸氢盐测序法检测30例MDS患者治疗前后骨髓DACT1基因甲基化状态、RTPCR检测治疗前后DACT1 mRNA及β-catenin mRNA变化。结果:治疗前MDS骨髓标本中,DACT1基因启动子区S1片段呈高甲基化状态,为15.21%,经青黄散联合健脾补肾方治疗后呈低甲基化状态,为5.45%(P<0.01)。治疗后DACT1 mRNA相对表达量由0.69±0.41升至1.83±2.01,较治疗前显著增加(P<0.01);β-catenin mRNA相对表达量治疗前后差异无统计学意义。结论:青黄散联合健脾补肾方通过DACT1基因去甲基化的作用效应,可能主要通过激活下游其他通路实现抑癌作用。

Objective:To investigate the effect of Qinghuang Powder combined with Jianpi Bushen Formula on DACT1 gene methylation and Wnt/β-catenin pathway in myelodysplastic syndrome(MDS)patients.Methods:The methylation of DACT1 gene in bone marrow of 30 MDS patients before and after treatment was detected by Bisulfite sequencing PCR(BSP),and the changes of DACT1 mRNA and β-catenin mRNA before and after treatment were detected by RT-PCR.Results:Before treatment,the S1 fragment of DACT1 gene promoter was hypermethylated(15.21%),which was hypomethylated(5.45%)after treatment with Qinghuang Powder and Jianpi Bushen Formula(P<0.01),and the relative expression of DACT1 mRNA increased from 0.69±0.41 to 1.83±2.01,which was significantly higher than that before treatment(P<0.01).There was no significant difference in the relative expression ofβ-catenin mRNA before and after treatment.Conclusion:Qinghuang Powder combined with Jianpi Bushen Formula has the effect of demethylation of DACT1 gene,possibly by activating other downstream pathways to achieve anti-cancer effect.