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本研究探讨abca5、mdr-1、kdr、dapk和irf-15个基因在CD7’急性髓细胞性白血病干/祖细胞的表达。利用实时定量RT-PCR(RQ-PCR)方法检测了15例正常骨髓(NBM)和16例AML患者骨髓单个核细胞(MNC)中5个基因的表达。采用流式细胞仪(FCM)分选8例NBM和21例AML患者骨髓CD34’CD38+祖细胞和CD34+CD38-干细胞,利用微量细胞RQ-PCR方法检测分选细胞中5个基因的表达。结果表明:NBMMNC均表达这5个基因,其中irf-1与dapk表达水平最高,相对表达量分别为4.08和3.86;abca5和mdr-1表达水平较低,为0.49和0.84;kdr表达最低为0.02。在经分选的CD34’CD38+祖细胞中,dapk和irf-1表达明显减低(p<0.05),kdr表达明显增加(P<0.05),其余2个基因无明显变化。在CD34’CD38-Lin-干细胞中,5个基因的表达均高于CD34+CD38+祖细胞,普遍增加至近2倍,而kdr增加了3倍,其中kdr和irf-1表达的增加具有统计学差异(P<0.05)。与NBM相比,AMLMNC中5个基因的表达水平均有不同程度减低,以abca5、mdr-1、kdr和dapk最为显著(p<0.05)。与AMLMNC相比,AMLCD34+CD38+细胞中,irf-1和dapk表达明显减低(p<0.05),其余3个基因表达增加,以kdr表达增加有统计学意义(p<0.05)。AMLCD34+CD38+与CD34+CD38-细胞比较,发现5个基因在CD34+CD38-细胞中的表达均增加,尤以kdr和irf-1的表达增加有意义(p<0.05)。AMLCD34+CD38-CD7+细胞与CD34’CD38-CD7-细胞中5个基因的表达均比CD34’CD38’细胞中的高,其中只有CD34’CD38-CD7+细胞中KDR和CD34’CD38-CD7-细胞中KDR和irf-1的表达增加并具有统计学差异(P<0.05)。结论:NBM中kdr主要表达在干/祖细胞中,而dapk和irf-1主要表达在较分化的细胞中。5个基因在干细胞阶段的表达均高于祖细胞阶段。AMLCD34+CD38-CD7+细胞与CD34+CD38-CD7-都具有干/祖细胞的基因表达特点。
This study was to investigate the expression of abca5, mdr-1, kdr, dapk and irf-15 genes in CD7 ’acute myeloid leukemia stem / progenitor cells. The expression of 5 genes in bone marrow mononuclear cells (MNC) from 15 normal bone marrow (NBM) and 16 AML patients was detected by real-time quantitative RT-PCR (RQ-PCR) Bone marrow CD34’CD38 + progenitor cells and CD34 + CD38- stem cells from 8 NBM patients and 21 AML patients were sorted by flow cytometry (FCM). The expression of 5 genes in sorted cells was detected by RQ-PCR. The results showed that: NBMMNC all expressed these five genes, of which irf-1 and dapk expression levels were highest, the relative expression levels were 4.08 and 3.86; abca5 and mdr-1 expression levels were low, 0.49 and 0.84; kdr expression was the lowest 0.02 . In sorted CD34’CD38 + progenitor cells, dapk and irf-1 expression was significantly decreased (p <0.05), kdr expression was significantly increased (P <0.05), the other two genes had no significant change. The expression of 5 genes in CD34’CD38-Lin- stem cells was higher than that of CD34 + CD38 + progenitors, generally increased nearly 2-fold and kdr increased 3-fold. The expression of kdr and irf-1 had statistical difference (P <0.05). Compared with NBM, the expression levels of 5 genes in AMLMNC decreased to some extent, and the most significant was abca5, mdr-1, kdr and dapk (p <0.05). Compared with AMLMNC, the expression of irf-1 and dapk in AMLCD34 + CD38 + cells was significantly decreased (p <0.05), the expression of the other three genes was increased, and the expression of kdr was increased statistically significant (p <0.05). Compared with CD34 + CD38- cells, AMLCD34 + CD38 + cells showed increased expression of all five genes in CD34 + CD38- cells, especially kdr and irf-1 (p <0.05). The expression of five genes in AMLCD34 + CD38-CD7 + cells and CD34’CD38-CD7- cells was higher than in CD34’CD38 ’cells, of which only KDR and CD34’CD38-CD7- cells in CD34’CD38-CD7 + cells The expression of KDR and irf-1 increased and had statistical difference (P <0.05). CONCLUSIONS: Kdr is primarily expressed in stem / progenitor cells in NBM, whereas dapk and irf-1 are predominantly expressed in more differentiated cells. The expression of 5 genes in stem cell stage were higher than in progenitor cell stage. Both AMLCD34 + CD38-CD7 + cells and CD34 + CD38-CD7- have the gene expression characteristics of stem / progenitor cells.