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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 investigated the effects of human umbilical cord mesenchymal stem cells (hUCMSCs) on the differentiation of HL-60 cells induced by all-trans retinoic acid (ATRA) and the proliferation of HL-60 cells. HL-60 cells were divided into 4 groups: control group without ATRA treatment, hucMSC group co-cultured with hucMSC cells, ATRA treated ATRA group and ATRA + hucMSC group treated with ATRA and co-cultured with hucMSC cells. Cell Counting Kit-8 (CCK8) was used to detect the proliferation of HL-60 cells in control group and hucMSC group. The morphological changes and NBT positive cells were observed under light microscope. The expression of c-myc gene was detected by real-time PCR CD11b surface markers were detected by flow cytometry to compare the differentiation of HL-60 cells in each group. The results showed that hucMSC cells could inhibit the proliferation of HL-60 cells in co-culture system, p <0.05 at 48 hours and p <0.01 at 72 hours after hucMSC: HL-60 was 1: P <0.05 at 72 hours. In ATRA + huc-MSC group, more HL-60 cells with neutrophilic morphology and higher NBT positive rate (p <0.05) appeared in ATRA + huc-MSC group compared with ATRA + hucMSC The expression of c-myc gene in HL-60 cells was lower (p <0.05) than that in HL-60 cells in ATRA + hucMSC group (p <0.05 for 48 hours and p <0.01 for 72 hours). Conclusion: Umbilical cord mesenchymal stem cells can inhibit the proliferation of HL-60 and enhance the differentiation of HL-60 cells induced by ATRA.