中药复方干预急性白血病干细胞FLT3和N-ras基因mRNA表达的研究
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摘要
目的:观察中药原方组及其拆方后的扶正组、祛邪组干预急性白血病干细胞FLT3和N-ras基因mRNA的表达,探讨中医药靶向干预急性白血病干细胞的作用机制。方法:将符合纳入标准的50例急性白血病患者,提取白血病干细胞,经过细胞培养后,将每一例细胞混悬液分成四组(空白组不加药物,实验组分别加入原方、扶正方、祛邪方中药制剂)。通过逆转录—聚合酶链式反应(RT-PCR),观察各组FLT3和N-ras基因mRNA半定量表达水平。结果:(1)原方组、扶正组、祛邪组FLT3和N-ras表达水平显著降低,与空白组比较均有非常显著性差异(P<0.01);原方组与扶正组、祛邪组表达水平比较有非常显著性差异(P<0.01);扶正组与祛邪组表达水平比较无显著性差异(P>0.05);FLT3和N-ras表达水平以空白组表达最强,原方组表达最弱,扶正组与祛邪组表达介于两者之间。(2)FAB亚型FLT3和N-ras基因表达水平比较无显著差异(P>0.05)。(3)FLT3和N-ras基因表达阳性患者的外周血白细胞数高(P<0.05),骨髓原始细胞比例高(P<0.05)。结论:中药原方能够降低FLT3和N-ras基因mRNA在急性白血病干细胞的表达水平,发现下调FLT3和N-ras基因表达为其可能作用机制,为中医药调控干预白血病干细胞的靶向治疗提供了可靠依据。
Objectives: To observe the expression of FLT3 and N-ras mRNA in Leukemic Stem Cell(LSC) by using the tranditional medicine fomular,and discuss the regulation mechanism of tranditional medicine fomular targeted intervention in LSC.Methods: We collected the bone marrow of the 50cases of AML,peripheral leukemic stem cell from human leukemic bone marrow,and culture the cell,the cell suspension was divided into four groups(the blank group without the drug,the experimental group,respectively,add the Yuanfang fomular,Fuzheng fomular,Quxie fomular).Through reverse transcription- polymerase chain reaction (RT-PCR), we observe the expression level of FLT3 and N-ras gene mRNA in LSC by using the tranditional medicine fomular. Results: (1)The expression of FLT3 and N-ras in Yuanfang fomular group were lower obviously than blank control group,Fuzheng fomular group and the Quxie fomular group(P<0.01);Yuanfang fomular can significantly reduce the expression of FLT3 and N-ras gene;There was no significant difference between the expression of Fuzheng fomular group and the Quxie fomular group(P>0.05), the strongest expression level is blank control group, the lowest expression level is Yuanfang fomular group, and the expression level of Fuzheng fomular group and the Quxie fomular group between the two groups.(2) There was no significant difference among the FAB subtypes (P>0.05).(3) The positive expression FLT3 and N-ras was associated with a higher white cell count(P<0.05),a higher percentage of bone marrow blast cells(P<0.05).Conclusion: The expression of FLT3 and N-ras mRNA in human leukemias is discreased by using the Yuanfang fomular. The possible mechanism is that lower FLT3 and N-ras gene expression, and provide a reliable basis for tranditional medicine fomular targeted intervention in LSC.
引文
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