RT-PCR法检测白血病干细胞Flt3和N-ras基因的表达

Study on the expression of Flt3 and N-ras genes in human leukemia stem cells using RT-PCR

目的:采用逆转录-聚合酶链反应(RT-PCR)法检测益气养阴方及其拆方后的扶正方组、祛邪方组急性白血病干细胞Flt3和N-ras基因的表达,以期阐明Flt3和N-ras基因表达与疾病发生、发展的关系。方法:RT-PCR法检测中药益气养阴方组、扶正方组、祛邪方组及对照组AML患者骨髓单个核细胞Flt3和N-ras基因表达的情况。结果:Flt3和N-ras基因表达阳性率益气养阴组、扶正组、祛邪组与对照组比较,差异有显著性意义(P均<0.01);益气养阴组与扶正组和祛邪组比较,差异有显著性意义(P均<0.01);扶正组与祛邪组比较,差异无显著性意义(P>0.05);各FAB亚型AML间Flt3与N-ras表达阳性率比较,差异无显著性意义(P>0.05)。结论:中药益气养阴方可降低Flt3和N-ras基因在AML细胞的表达水平,抑制AML细胞的克隆性增殖,对AML具有治疗作用。

Objective：To investigate the expression of Flt3 and N-ras genes in different TCM prescription-treated acute myeloid leukemia （AML） stem cells, in an attempt to reveal the relationship between the genes and the development of AML. Methods： Reverse transcriptase polymerase chain reaction was used to detect the mRNA expressions of Flt3 and N-ras in marrow mononuclear ceils from AML patients. Three kinds of TCM fnmular were used. Results： Compared with the control group,leukemia stem cells were more sensitive to the Yiqiyangyin, Fuzheng and Quxie fomulars （P 〈 0.01 ） in regard to the expression of Flt3 and N-ras on the level of mRNA. The positive rate of Fh3 and N-ras mRNA in Yiqiyangyin fomular was less than that in the Fuzheng and Quxie fomulars（ P 〈0.01 ） and no statistically significant difference between the Fuzheng and Quxie fomulars（ P 〉 0.05 ）. No significant difference in Flt3 and N-ras expression was noted among AML subtypes in all formular-treated groups（ P 〉 0.05 ）. Conclusion ：The yiqiyangyin fomular could inhibit the proliferation of leukemia cells by decreasing the expression of Fit3 and Nras gene on the cellular level and may have therapeutic effects on AML.