人脐带间充质干细胞增强全反式维甲酸对HL-60细胞的诱导分化作用  

Enhancement of All-trans Retinoic Acid Induced HL-60 Leukemia Cell Differentiation by Human Umbilical Cord Mesenchymal Stem Cells

作　　者：吴昊 马凤霞 李建平 杨少光 贾海蓉 卢世红 任倩 赵钦军 刑文 张磊[1] 韩忠朝 

机构地区：[1]中国医学科学院、北京协和医学院血液学研究所、血液病医院实验血液学国家重点实验室,天津300020

出　　处：《中国实验血液学杂志》2010年第4期877-881,共5页Journal of Experimental Hematology

基　　金：国家自然科学基金项目(编号30872330);国家自然科学基金项目(编号30900557)

摘　　要：本研究探讨人脐带间充质干细胞(hucMSC)对全反式维甲酸(ATRA)诱导HL-60细胞分化以及对HL-60增殖的影响。HL-60细胞分为4组:未经ATRA处理的对照组、与hucMSC细胞共培养的hucMSC组,经ATRA处理的ATRA组以及经ATRA处理并与hucMSC细胞共培养的ATRA+hucMSC组。在规定时间采用Cell Counting Kit-8(CCK8)检测对照组和hucMSC组HL-60细胞增殖情况;在光学显微镜下观察各组细胞形态、NBT阳性细胞;应用real-timePCR方法检测c-myc基因表达水平以及流式细胞术检测CD11b表面标志以比较各组中HL-60细胞的分化情况。结果表明:共培养体系中hucMSC细胞能抑制HL-60的增殖,hucMSC∶HL-60为1∶1时,48小时时p<0.05,72小时时p<0.01;hucMSC∶HL-60为1∶5时,72小时时p<0.05。在2μmol/LATRA的刺激下,ATRA+huc-MSC组与ATRA组相比,出现更多具有中性粒细胞形态的HL-60细胞和更高的NBT阳性率(p<0.05);ATRA+hucMSC组中HL-60细胞c-myc基因表达更低(p<0.05);ATRA+hucMSC组中HL-60细胞CD11b的表达更高(48小时p<0.05,72小时p<0.01)。结论:脐带间充质干细胞能抑制HL-60增殖并且增强ATRA对HL-60细胞的诱导分化作用。This study was aimed to investigate the enhancement of all-trans retinoic acid-induced HL-60 leukemia cell differentiation by human umbilical cord mesenchymal stem cells （hucMSC）. The HL-60 cells were divided into 4 groups： control group （HL-60 cells treated without ATRA）, hucMSC group （HL-60 cells co-cultured with hucMSCs）, ATRA group （HL-60 cells treated with ATRA） and ATRA ＋ hucMSC group （HL-60 cells treated with ATRA and co- cultcured with hucMSCs）. The proliferations of control group and hucMSC group were compared by Cell Counting Kit- 8 （ CCK8 ）. The morphology of HL-60 cells and NBT positive rate in 4 groups were observed and compared by means of microscopy, the c-myc expression of HL-60 cells in different groups was evaluated by real-time PCR, and the CDllb expression on HL-60 cells in different groups were detected by flow cytometry. The results showed that in the co-culturing system, hucMSCs could inhibit the proliferation of HL-60 （ hucMSC： HL-60 is 1 ： 1, 48 hours p 〈 0.05, 72 hours p 〈 0.01 ; hucMSC： HL-60 is 1 ： 5, 72 hours p 〈 0.05 ）. In condition of stimulation with 2 μmol/L ATRA, the neutrophil like HL-60 cells and NBT positive rate in ATRA ＋ hucMSC group were higher than those in ATRA group （p 〈 0.05 ）. The c-myc expression of HL-60 cells in ATRA ＋ hucMSC group was lower than that in ATRA group （p 〈0.05 ）. Furthermore, HL-60 cells in ATRA ＋ hucMSC group had stronger CD1 lb expression than ATRA group （48 hours p 〈 0.05, 72 hours p 〈0.01 ）. It is concluded that hucMSC not only can inhibit the proliferation of HL-60 cells, but also can enhance the differentiation effect of HL-60 cells induced by ATRA.

关 键 词：脐带间充质干细胞 HL-60 全反式维甲酸 诱导分化 中性粒细胞 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学] Q253[医药卫生—基础医学]