姜黄素下调CyclinD1和Bcl-2的表达增强急性髓系LSCs对柔红霉素的敏感性

目的 探讨姜黄素(CUR)增强急性髓系白血病干细胞(LSCs)CD34+ CD38-KG1a细胞对柔红霉素(DNR)敏感性的机制.方法 流式细胞术分析KG1a细胞的CD34、CD38表面分子的表达情况.四甲基偶氮唑蓝(MTT)法获取CUR作用CD34+CD38-KG1a细胞的IC50 .M T T 法、甲基纤维素克隆形成实验和流式细胞术分别检测DNR对两种细胞(CD34+ CD38-KG1a和CUR/CD34+CD38-KG1a)的增殖抑制作用、克隆形成能力和凋亡影响.逆转录PCR(RT-PCR)检测细胞Bcl-2、Bax和XIAP的mRNA表达.Western blot分析细胞周期蛋白D1(CyclinD1)、Bcl-2、Bax 和 XIAP蛋白表达.结果 KG1a细胞系的CD34+CD38-KG1a细胞比例为(98 .2 ± 3 .2)% .CUR作用CD34+CD38-KG1a细胞24 h的IC50 =100 μmol/L.中、高浓度(0 . 8、2 .0 μg/mL)DNR对CUR/CD34+CD38-KG1a细胞的增殖抑制作用比CD34+CD38-KG1a细胞强(P＜0 .05).DNR对CUR/CD34+CD38-KG1a细胞克隆形成能力的抑制作用较CD34+ CD38-KG1a细胞强(P＜0 .05).各DNR浓度组中 ,CUR/CD34+CD38-KG1a细胞凋亡率均比CD34+CD38-KG1a细胞高(P＜0 .05).Bcl-2的mRNA及CyclinD1和Bcl-2蛋白表达下降.Bax、XIAP的mRNA和蛋白表达变化不明显.结论 CUR能增强CD34+ CD38-KG1a细胞对DNR的敏感性 ,与CUR下调CD34+CD38-KG1a细胞CyclinD1和Bcl-2的表达相关.

Curcumin enhancing sensitivity of acute myeloid leukemia stem cells to daunorubicin by down-regulating expression of CyclinD1 and Bcl-2

Objective To explore the potential mechanism of curcumin(CUR) enhancing the sensitivity of acute myeloid leu-kemistem cells(LSCs) CD34+CD38-KG1cellto daunorubicin (DNR) .MethodThe expression of surface moleculeCD34 , CD38 in KG1cellwadetected by the flow cytometry .The half maximal inhibitory concentration (IC50 ) of curcumin to CD34+CD38-Kglcellwacalculated by the Mtmethod .MT,methylcellulose colony formation assay and flow cytometry were ap-plied to examine the effectof Dnon the proliferation ,clone forming ability and apoptosiin the two kindof cell(CD34+CD38-KG1celland CUR/CD34+CD38-KG1cells) respectively .The mRNA-expression of Bcl-2 ,Bax and XIAP in the two cellwere analyzed by RT-PC.Meanwhile ,the protein-expression of CyclinD1 ,Bcl-2 ,Bax and XIAP were detected by Western Blo.ResultThe percentage of CD34+CD38-KG1in KG1cell line wa(98 .2 ± 3 .2)% .The IC50 of curcumin treating CD34+CD38-KG1fo24 h wa100 μmol/L .The inhibition effecof high and middle concentration(0 .8 ,2 .0 μg/mL) of Dnon the proliferation of CUR/CD34+CD38-KG1cellwastrongethan thaof CD34+CD38-KG1cell(P＜0 .05) .The inhibitory role of Dnto the colony formation on CUR/CD34+CD38-KG1cellwamore effective than thaof CD34+ CD38-KG1cells(P＜0 .05) .The ap-optotirate of CUR/CD34+ CD38- KG1cellin each concentration group of Dnwahighethan thaof CD 34+ CD38- KG1cells(P＜0 .05) .The mRNA-expression of Bcl-2 and the protein-expression of Bcl-2 and CyclinD1 were down-regulated .Howeve, no significanchange waobserved aboth mRNA-expression and protein-expression of Bax and XIAP .Conclusion Cucan en-hance the sensitivity of CD34+CD38-KG1cellto DN,which mighbe associated with down-regulating the expression of Cy-clinD1and Bcl-2 .