【摘要】 目的:观察吲哚美辛对急变期慢性粒细胞白血病患者骨髓CD34~+细胞增殖和自我更新能力的影响,并从Wnt/β-catenin信号通路的角度初步探讨吲哚美辛作用的分子机制。方法:采用免疫磁珠分选技术分选获得慢性期和急变期慢性粒细胞白血病患者骨髓以及健康人脐带血标本中的CD34~+细胞,并采用FCM和实时荧光定量PCR法检测CD34~+细胞中β-catenin和Bcr-Abl的表达水平。用吲哚美辛或(和)伊马替尼处理CD34~+细胞后,采用FCM法检测细胞中β-catenin的表达水平,MTT法检测细胞增殖情况,克隆形成实验检测细胞克隆形成能力,实时荧光定量PCR法检测c-Myc和cyclin D1的mRNA水平。结果:成功分选出各标本中的CD34~+细胞,其中急变期慢性粒细胞白血病患者的CD34~+细胞中β-catenin和Bcr-Abl均高表达(P值均<0.05)。吲哚美辛作用后,急变期慢性粒细胞白血病患者的CD34~+细胞中β-catenin表达显著下调(P<0.05),细胞增殖和克隆形成能力均被明显抑制(P值均<0.05),并且β-catenin下游靶基因c-Myc和cyclin D1的mRNA水平也被明显下调(P值均<0.05);而吲哚美辛与伊马替尼联合用药后,以上作用效果更加明显(P值均<0.05)。结论:吲哚美辛可能通过影响Wnt/β-catenin信号通路,抑制CD34~+细胞的增殖和自我更新能力,从而增强伊马替尼对急变期慢性粒细胞白血病患者中CD34~+细胞的杀伤力。更多还原

【Abstract】 Objective :To investigate the effect of indomethacin on the proliferation and self-renewal ability of CD34~+ cells from bonemarrow of the patients with blastic-phase chronic myeloid leukemia,and to explore its molecular mechanism related to Wnt/β-catenin signal pathway.Methods:The immune magnetic bead separation was used to sort CD34~+ cells from bone marrow of the patients with blastic-phase chronic myeloid leukemia or chronic-phase chronic myeloid leukemia and from umbilical cord blood of the healthy volunteers.The protein and mRNA expressions of β-catenin and Bcr-Abl in these CD34~+ cells were analyzed by FCM and real-time fluorescent quantitative PCR(RFQ-PCR),respectively.After CD34~+ cells were treated with indomethacin or/and imatinib,the protein level of β-catenin was detected by FCM,the cell proliferation was assessed by MTT assay,the colony-forming ability was evaluated by colony-forming assay,and the mRNA levels of c-Myc and cyclin D1 were detected by RFQ-PCR.Results:CD34~+ cells were successfully acquired from the different samples.The protein and mRNA expressions of β-catenin and Bcr-Abl in the CD34~+ cells from blastic-phase chronic myeloid leukemia samples were markedly higher than those from other samples(all P < 0.05).Indomethacin significantly suppressed the expression of β-catenin,cell proliferation and colony-forming ability of CD34~+ cells from blastic-phase chronic myeloid leukemia samples(all P < 0.05),especially combined with imatinib(all P < 0.05).In addition,indomethacin decreased the mRNA levels of β-catenin-targeted genes c-Myc and cyclin D1 in CD34~+ cells from blastic-phase chronic myeloid leukemia samples as compared with the other samples(all P < 0.05),especially combined with imatinib(all P < 0.05).Conclusion:Indomethacin significantly suppresses the cell proliferation and colony-forming ability of CD34~+ cells from blastic-phase chronic myeloid leukemia samples,and enhances the effect of imatinib on those CD34~+cells.Wnt/β-catenin signal pathway may be involved in this process.更多还原